CD14
Monocyte differentiation antigen CD14 precursor (Myeloid cell-specific leucine-rich glycoprotein) (CD14 antigen) [Contains: Monocyte differentiation antigen CD14, urinary form; Monocyte differentiation antigen CD14, membrane-bound form]
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Mononuclear phagocytes and NK cells constitute the first line of innate immune defense. How these cells interact and join forces against cancer is incompletely understood. Here, we observed an early accumulation of slan (6-sulfo LacNAc) non-classical monocytes (slanMo) in stage I melanoma, which was followed by an increase in NK cell numbers in stage III. Accordingly, culture supernatants of slanMo induced migration of primary human NK cells [i]in vitro[/i] via the chemotactic cytokine IL-8 (CXCL8), suggesting a role for slanMo in NK cell recruitment into cancer tissues. High levels of TNF-α and IFN-γ were produced in co-cultures of TLR-ligand stimulated slanMo and NK cells, whereas much lower levels were contained in cultures of slanMo and NK cells alone. Moreover, TNF-α and IFN-γ concentrations in slanMo/NK cell co-cultures exceeded those in CD14 monocyte/NK cell and slanMo/T cell co-cultures. Importantly, TNF-α and IFN-γ that was produced in TLR-ligand stimulated slanMo/NK cell co-cultures induced senescence in different melanoma cell lines, as indicated by reduced melanoma cell proliferation, increased senescence-associated β-galactosidase expression, p21 upregulation, and induction of a senescence-associated secretory phenotype (SASP). Taken together, we identified a role for slanMo and NK cells in a collaborative innate immune defense against melanoma by generating a tumor senescence-inducing microenvironment. We conclude that enhancing the synergistic innate immune crosstalk of slanMo and NK cells could improve current immunotherapeutic approaches in melanoma.
Keywords
- NK cell
- cytokines
- melanoma
- senescence
- slanMo
Intestinal (i.e., "gut") permeability may be related to cardiovascular disease (CVD) risk, but biomarkers for gut permeability are limited and associations with CVD risk are unknown-particularly among older adults. This cross-sectional study aimed to determine if serum biomarkers related to gut permeability [intestinal fatty acid-binding protein (iFABP)] and bacterial toxin clearing [cluster of differentiation 14 (CD14), lipopolysaccharide binding protein (LBP)] are associated with CVD risk among older adults. Older adults (n = 74, 69.6 ± 6.5-years-old) were stratified by CVD risk category. One-way ANOVAs determined differences in each biomarker by risk category, and associations with risk score were evaluated with Pearson correlations. LBP (p = 0.007), but not iFABP and CD14, was significantly different between CVD risk categories. Post-hoc tests indicated LBP was higher in moderate risk and high-moderate risk compared to the high risk category (p < 0.005). Evaluation of LBP and individual components in the risk score demonstrated a moderate, negative correlation of LBP with age and systolic blood pressure (r = - 0.335 and r = - 0.297) and a small positive correlation between LBP and total cholesterol and LDL cholesterol (r = 0.204 and r = 0.220). Lower risk for CVD was associated with higher circulating concentrations of LBP, lower iFABP, and lower systemic inflammation in older adults. Further, there were small positive relationships between total and LDL cholesterol and circulating levels of LBP. These data suggest LBP may be a key component in reducing CVD risk in older adults.
Keywords
- Aging
- Cardiovascular disease risk
- Intestinal permeability
- Lipopolysaccharide binding protein
It is well established that multinucleation is central for osteoclastic bone resorption. However, our knowledge on the mechanisms regulating how many nuclei an osteoclast will have is limited. The objective of this study was to investigate donor-related variations in the fusion potential of in vitro-generated osteoclasts. Therefore, CD14 monocytes were isolated from 49 healthy female donors. Donor demographics were compared to the in vivo bone biomarker levels and their monocytes' ability to differentiate into osteoclasts, showing that: (1) C-terminal telopeptide of type I collagen (CTX) and procollagen type I N-terminal propeptide (PINP) levels increase with age, (2) the number of nuclei per osteoclast in vitro increases with age, and (3) there is a positive correlation between the number of nuclei per osteoclast in vitro and CTX levels in vivo. Furthermore, the expression levels of the gene encoding dendritic cell-specific transmembrane protein ([i]DCSTAMP[/i]) of osteoclasts in vitro correlated positively with the number of nuclei per osteoclast, CTX levels in vivo, and donor age. Our results furthermore suggest that these changes in gene expression may be mediated through age-related changes in DNA methylation levels. We conclude that both intrinsic factors and age-induced increase in fusion potential of osteoclasts could be contributing factors for the enhanced bone resorption in vivo, possibly caused by increased expression levels of [i]DCSTAMP[/i].
Keywords
- CTX
- DC-STAMP
- DNA methylation
- aging
- cell fusion
- epigenetics
- menopause
- multinucleation
- osteoclast
- osteoclastogenesis
Soluble toll-like receptor-2 (sTLR2) and soluble CD14 (sCD14) in saliva are defense proteins that bind specific microbe-associated molecular patterns. Since the oral flora changes with aging, the objective of this study is to determine and compare the concentration of sTLR2 and sCD14 in the saliva of healthy individuals in age groups from the first to the sixth decade of life. Unstimulated whole saliva was collected after obtaining informed consent. The concentration of sCD14 and sTLR-2 was measured by enzyme-linked immunosorbent assay. Statistical differences between the age groups were determined by analysis of variance. The relationship between the two markers in each age group was evaluated by Pearson's correlation coefficient and linear regression analyses. The concentration of salivary sTLR2 was highest in the youngest, and that of the sCD14 was highest in the oldest age group. While the salivary sCD14 and the sTLR2 exhibited a moderate negative correlation in the youngest, the relationship between the two markers was inversed in the oldest age group. The results of our exploratory study suggest a need to adjust for age-dependent changes in sCD14 and sTLR2 in healthy saliva while assessing the two proteins as biomarkers.
MeSH Terms
- Adolescent
- Adult
- Aging
- Biomarkers
- Child
- Child, Preschool
- Humans
- Lipopolysaccharide Receptors
- Middle Aged
- Saliva
- Salivary Proteins and Peptides
- Toll-Like Receptor 2
- Young Adult
Keywords
- Age changes
- CD14
- Saliva
- Toll-like receptor-2
To test the hypothesis that the inflammatory marker plasma soluble CD14 (sCD14) associates with incident dementia and related endophenotypes in 2 community-based cohorts. Our samples included the prospective community-based Framingham Heart Study (FHS) and Cardiovascular Health Study (CHS) cohorts. Plasma sCD14 was measured at baseline and related to the incidence of dementia, domains of cognitive function, and MRI-defined brain volumes. Follow-up for dementia occurred over a mean of 10 years (SD 4) in the FHS and a mean of 6 years (SD 3) in the CHS. We studied 1,588 participants from the FHS (mean age 69 ± 6 years, 47% male, 131 incident events) and 3,129 participants from the CHS (mean age 72 ± 5 years, 41% male, 724 incident events) for the risk of incident dementia. Meta-analysis across the 2 cohorts showed that each SD unit increase in sCD14 was associated with a 12% increase in the risk of incident dementia (95% confidence interval 1.03-1.23; [i]p[/i] = 0.01) following adjustments for age, sex, [i]APOE[/i] ε4 status, and vascular risk factors. Higher levels of sCD14 were associated with various cognitive and MRI markers of accelerated brain aging in both cohorts and with a greater progression of brain atrophy and a decline in executive function in the FHS. sCD14 is an inflammatory marker related to brain atrophy, cognitive decline, and incident dementia.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Atrophy
- Biomarkers
- Brain
- Cognitive Dysfunction
- Dementia
- Female
- Humans
- Incidence
- Lipopolysaccharide Receptors
- Longitudinal Studies
- Male
- Middle Aged
Fracture repair is initiated by a multitude of immune cells and induction of an inflammatory cascade. Alterations in the early healing response due to an aged adaptive immune system leads to impaired bone repair, delayed healing or even formation of non-union. However, immuno-senescence is not limited to the adaptive immunity, but is also described for macrophages, main effector cells from the innate immune system. Beside regulation of pro- and anti-inflammatory signaling, macrophages contribute to angiogenesis and granulation tissue maturation. Thus, it seems likely that an altered macrophage function due to aging may affect bone repair at various stages and contribute to age related deficiencies in bone regeneration. To prove this hypothesis, we analyzed the expression of macrophage markers and angiogenic factors in the early bone hematoma derived from young and aged osteotomized Spraque Dawley rats. We detected an overall reduced expression of the monocyte/pan-macrophage markers CD14 and CD68 in aged rats. Furthermore, the analysis revealed an impaired expression of anti-inflammatory M2 macrophage markers in hematoma from aged animals that was connected to a diminished revascularization of the bone callus. To verify that the age related disturbed bone regeneration was due to a compromised macrophage function, CD14 macrophage precursors were transplanted locally into the osteotomy gap of aged rats. Transplantation rescued bone regeneration partially after 6 weeks, demonstrated by a significantly induced deposition of new bone tissue, reduced fibrosis and significantly improved callus vascularization.
MeSH Terms
- Age Factors
- Aging
- Animals
- Antigens, CD
- Antigens, Differentiation, Myelomonocytic
- Biomarkers
- Bone Regeneration
- Bone and Bones
- Female
- Fractures, Bone
- Gene Expression
- Lipopolysaccharide Receptors
- Macrophages
- Osteotomy
- Rats, Sprague-Dawley
- Wound Healing
Keywords
- CD14 cells
- aging
- angiogenesis
- bone regeneration
- compromised healing
- macrophage
- monocyte
Stromal mesenchymal stem cells (MSCs) have the capability to self-renew and can differentiate into multiple cell types of the mesoderm germ layer, but their properties are affected by molecular aging mechanisms. MSCs can be obtained from adipose tissue termed as adipose-derived stem/stromal cells (ASCs) representing a promising tool for studying age-related diseases in detail. ASCs from young (16 weeks) and old (>108 weeks) rabbits were successfully isolated and propagated. ASCs showed the typical morphology and stained positive for CD105, Vimentin, Collagenase 1A, and negative for CD14, CD90, and CD73, demonstrating their mesenchymal origin. ASCs expressed MSC markers, including [i]MYC[/i], [i]KLF4[/i], [i]CHD1[/i], [i]REST[/i], and [i]KAT6A[/i], whereas pluripotency-related genes, such as [i]NANOG[/i], [i]OCT4[/i], and [i]SOX2[/i], were not expressed. Aged ASCs showed altered protein and mRNA levels of APOE, ATG7, FGF2, PTEN, and SIRT1. Adipogenic differentiation of old visceral ASCs was significantly decreased compared with young visceral ASCs. We successfully established rabbit ASC cultures representing an [i]in vitro[/i] model for the analysis of stem cell aging mechanisms. ASCs, obtained from old female rabbits, showed age- and source-specific alteration due to aging of the donor. Stem cell plasticity was altered with age as shown by reduced adipogenic differentiation capacity.
MeSH Terms
- Adipogenesis
- Adipose Tissue
- Aging
- Animals
- Biomarkers
- Cell Differentiation
- Cell Plasticity
- Cell Proliferation
- Cells, Cultured
- Female
- Mesenchymal Stem Cells
- Rabbits
Keywords
- adipogenic differentiation
- adipose-derived stem/stromal cells
- aging biomarkers
- and stem cell plasticity
- healthy aging
The key to newer therapeutic and eradication approaches often lies in understanding slow disease progression in HIV infection. The paediatric population has been poorly studied in this regard. We aimed to describe a cohort of perinatally infected long-term nonprogressor (LTNP) children living with HIV in India and to evaluate the immune biomarkers of disease progression. LTNPs (ART-naïve, with a CD4 count ≥ 500 cells/μL at age ≥ 7 years) among the cohort of HIV-infected children were identified and monitored longitudinally, and their CD4 T-cell counts and plasma viral loads were measured every 6 months. The plasma monocyte/macrophage activation markers, namely soluble CD14 (sCD14), soluble CD163 (sCD163) and interferon-inducible protein-10 (IP-10) were measured by enzyme-linked immunosorbent assay (ELISA) in LTNPs and progressors. The Mann-Whitney U-test was used to compare the two groups and P values < 0.05 were considered statistically significant. Spearman's rank or Pearson's correlation coefficient (r) was calculated to determine the associations between variables. Among 378 children living with HIV-1 surveyed in our cohort, 40 (10.6%) were LTNPs. Longitudinal analysis of the LTNP data showed that both CD4 count and viral load declined significantly with age (P < 0.0001 for both). Plasma sCD14 levels were significantly (P < 0.005) higher in progressors and sCD163 levels were significantly (P < 0.0001) higher in LTNPs. The prevalence of LTNPs in our cohort of perinatally infected children living with HIV was 10.6%. We observed a trend for associations between the increasing sCD163 monocyte/macrophage activation marker levels, declining CD4 counts and the gradual loss of nonprogressor status with age in the LTNPs. These findings underscore the need for early antiretroviral therapy in those children with proven slow disease progression.
MeSH Terms
- Adolescent
- Aging
- Antigens, CD
- Antigens, Differentiation, Myelomonocytic
- Biomarkers
- CD4 Lymphocyte Count
- Chemokine CXCL10
- Child
- Cross-Sectional Studies
- Disease Progression
- Female
- HIV Infections
- HIV Long-Term Survivors
- HIV-1
- Humans
- India
- Infectious Disease Transmission, Vertical
- Longitudinal Studies
- Male
- Monocytes
- Prevalence
- Receptors, Cell Surface
- Viral Load
Keywords
- HIV-1
- children
- immune biomarkers
- long-term nonprogressors
- monocyte activation
To elucidate biochemical mechanisms leading to seed deterioration, we studied 23 wheat genotypes after exposure to seed bank storage for 6-16 years compared to controlled deterioration (CD) at 45 °C and 14 (CD14) and 18% (CD18) moisture content (MC) for up to 32 days. Under two seed bank storage conditions, seed viability was maintained in cold storage (CS) at 0 °C and 9% seed MC, but significantly decreased in ambient storage (AS) at 20 °C and 9% MC. Under AS and CS, organic free radicals, most likely semiquinones, accumulated, detected by electron paramagnetic resonance, while the antioxidant glutathione (GSH) was partly lost and partly converted to glutathione disulphide (GSSG), detected by HPLC. Under AS the glutathione half-cell reduction potential (E ) shifted towards more oxidising conditions, from -186 to -141 mV. In seeds exposed to CD14 or CD18, no accumulation of organic free radicals was observed, GSH and seed viability declined within 32 and 7 days, respectively, GSSG hardly changed (CD14) or decreased (CD18) and E shifted to -116 mV. The pH of extracts prepared from seeds subjected to CS, AS and CD14 decreased with viability, and remained high under CD18. Across all treatments, E correlated significantly with seed viability ([i]r[/i] = 0.8, [i]p[/i]<.001). Data are discussed with a view that the cytoplasm is in a glassy state in CS and AS, but during the CD treatments, underwent transition to a liquid state. We suggest that enzymes can be active during CD but not under the seed bank conditions tested. However, upon CD, enzyme-based repair processes were apparently outweighed by deteriorative reactions. We conclude that seed ageing by CD and under seed bank conditions are accompanied by different biochemical reactions.
MeSH Terms
- Antioxidants
- Disulfides
- Glutathione
- Hydrogen-Ion Concentration
- Oxidation-Reduction
- Seeds
- Sulfhydryl Compounds
- Time Factors
- Triticum
Keywords
- Antioxidants
- EPR
- ESR
- gene bank
- germination
- organic radicals
- seed longevity
The incidence of cardiovascular disease is higher in HIV-positive (HIV ) patients than it is in the average population, and combination antiretroviral therapy (cART) is a recognized risk factor for cardiovascular disease. However, the molecular mechanisms that link cART and cardiovascular disease are currently unknown. Our study explores the role of the activation of p90RSK, a reactive oxygen species-sensitive kinase, in engendering senescent phenotype in macrophages and accelerating atherogenesis in patients undergoing cART. Peripheral whole blood from cART-treated HIV individuals and nontreated HIV-negative individuals was treated with H O (200 µmol/L) for 4 minutes, and p90RSK activity in CD14 monocytes was measured. Plaque formation in the carotids was also analyzed in these individuals. Macrophage senescence was determined by evaluating their efferocytotic ability, antioxidation-related molecule expression, telomere length, and inflammatory gene expression. The involvement of p90RSK-NRF2 signaling in cART-induced senescence was assessed by p90RSK-specific inhibitor (FMK-MEA) or dominant-negative p90RSK (DN-p90RSK) and NRF2 activator (NRF2A). Further, the severity of atherosclerosis was determined in myeloid cell-specific wild-type and DN-p90RSK transgenic mice. Monocytes from HIV patients exhibited higher levels of p90RSK activity and were also more sensitive to reactive oxygen species than monocytes from HIV-negative individuals. A multiple linear regression analysis involving cART, Reynolds cardiovascular risk score, and basal p90RSK activity revealed that cART and basal p90RSK activity were the 2 significant determinants of plaque formation. Many of the antiretroviral drugs individually activated p90RSK, which simultaneously triggered all components of the macrophage senescent phenotype. cART inhibited antioxidant response element reporter activity via ERK5 S496 phosphorylation. NRF2A reversed the H O -induced overactivation of p90RSK in cART-treated macrophages by countering the induction of senescent phenotype. Last, the data obtained from our gain- or loss-of-function mice conclusively showed the crucial role of p90RSK in inducing senescent phenotype in macrophages and atherogenesis. cART increased monocyte/macrophage sensitivity to reactive oxygen species- in HIV individuals by suppressing NRF2-ARE activity via p90RSK-mediated ERK5 S496 phosphorylation, which coordinately elicited senescent phenotypes and proinflammatory responses. As such, our report underscores the importance of p90RSK regulation in monocytes/macrophages as a viable biomarker and therapeutic target for preventing cardiovascular disease, especially in HIV patients treated with cART.
MeSH Terms
- Animals
- Anti-Retroviral Agents
- Cellular Senescence
- Female
- HIV Seropositivity
- HIV-1
- Humans
- Macrophages
- Male
- Mice
- NF-E2-Related Factor 2
- Oxidative Stress
- Protein Kinase Inhibitors
- Ribosomal Protein S6 Kinases, 90-kDa
Keywords
- HIV
- antioxidants
- atherosclerosis
- reactive oxygen species
- senescence
- telomere
There is evidence that pepsin can aggravate tonsil hypertrophy. Pepstatin is a potent inhibitor of pepsin activity and could protect patients against reflux tonsil hypertrophy by inhibiting pepsin. We examined the effects of pepstatin on the development of tonsil hypertrophy to investigate pepsin's role in the pathogenesis of tonsil lesions. We investigated whether pepstatin suppresses pepsin-mediated lymphocyte proliferation in tonsil hypertrophy. Forty-nine children with tonsil hypertrophy and twenty-two adults with tonsillitis were recruited to the study prior to surgery. Tonsil tissue from each patient was harvested and assessed for changes in the number of lymphocytes and macrophages in the presence of pepsin and pepstatin. We found that the proportions of CD4- and CD14-positive cells were significantly lower (p < 0.05), but that the proportions of CD19- and CD68-positive cells were significantly higher (p < 0.05), in children than in adults. There were significantly more CD4-positive cells after pepsin treatment, but these numbers were reduced by pepstatin. The levels of both interleukin-2 (IL-2) and interferon gamma (IFN-γ) increased significantly in response to pepsin, but were reduced when pepsin was inhibited by pepstatin. The level of IL-10 is reduced in pepsin-treated CD4 cells and the level is restored by pepstatin. IL-2 blocking reduced the increased CD4 cell number by pepsin. But, an additive or a synergic effect is not founded in combined with IL-2 blocking and pepstatin. Pepsin-positive cells did not co-localize with CD20 and CD45 cells, but they were found surrounding CD20- and CD45-positive hypertrophic tonsil cells. Pepsin-positive cells co-localized with CD68-positive cells. It is probable that pepsin from extraesophageal reflux aggravates tonsil hypertrophy and pepstatin exerts a protective effect by inhibiting pepsin activity.
MeSH Terms
- Adolescent
- Adult
- Aging
- Child
- Child, Preschool
- Female
- Humans
- Hypertrophy
- In Vitro Techniques
- Interferon-gamma
- Interleukin-10
- Interleukin-2
- Lymphocytes
- Macrophages
- Male
- Palatine Tonsil
- Pepsin A
- Pepstatins
- Pharyngeal Diseases
This study aimed to evaluate whether ICU patients who developed persistent critical illness displayed an immune profile similar to an aged immune phenotype and any associations with patient outcomes. Twenty two critically ill ICU patients (27-76 years, 15 males), at day 5 of mechanical ventilation, and 22 healthy age-matched controls (27-77 years, 13 males) were recruited. Frequency and phenotype of innate and adaptive immune cells and telomere length in peripheral blood mononuclear cells (PBMCs) were measured. An elevated granulocyte count (p < 0.0001), increased numbers of immature granulocytes (p < 0.0001), increased CD16 monocytes (p = 0.003) and CD14 HLADR monocytes (p = 0.004) and lower NK cell numbers (p = 0.007) were observed in ICU patients compared to controls. Critically ill patients also had lower numbers of total T lymphocytes (p = 0.03), naïve CD4 T cells (p = 0.003) and PTK7 recent thymic emigrants (p = 0.002), and increased senescent CD28 CD57 CD4 T cells (p = 0.02), but there was no difference in PBMC telomere length. Regulatory immune cell frequency was affected with reduced circulating CD19 CD24 CD38 regulatory B cells (p = 0.02). However, only a raised neutrophil:lymphocyte ratio and reduced frequency of CD14 HLADR monocytes were associated with poor outcomes. We conclude that persistent critical illness results in changes to immune cell phenotype only some of which are similar to that seen in physiological ageing of the immune system.
MeSH Terms
- Adult
- Aged
- Aging
- Critical Illness
- Female
- Healthy Volunteers
- Humans
- Immunity, Cellular
- Intensive Care Units
- Leukocyte Count
- Leukocytes, Mononuclear
- Male
- Middle Aged
- Phenotype
- Telomere Homeostasis
Endotoxemia-induced inflammation has been associated with insulin resistance and atherosclerosis, ultimately increasing the risk of coronary heart disease. Increased intestinal permeability is an important event leading to endotoxemia. This study aims to elucidate the possible association between endotoxin (lipopolysaccharide) and zonulin (a biomarker of intestinal permeability) levels and the risk of coronary heart disease, and thus healthy aging. Serum levels of zonulin, lipopolysaccharide and soluble CD14 (a protein that binds lipopolysaccharide) were measured in disease-free centenarians, young healthy controls and patients with precocious acute myocardial infarction. Disease-free centenarians had significantly lower levels of serum zonulin ([i]P[/i]<0.01) and lipopolysaccharide ([i]P[/i]<0.001) than young patients with acute myocardial infarction, and had significantly lower concentrations of serum lipopolysaccharide than young healthy controls ([i]P[/i]<0.05). No significant differences were found for soluble CD14 between groups. Our findings may stimulate further research into the role played by intestinal permeability and endotoxemia not only in coronary heart disease but also in lifespan modulation.
Keywords
- centenarians
- endotoxemia
- inflammation
- longevity
Neutrophils, basophils, and monocytes are continuously produced in bone marrow via myelopoiesis, circulate in blood, and are eventually removed from circulation to maintain homeostasis. To quantitate the kinetics of myeloid cell movement during homeostasis, we applied 5-bromo-2'-deoxyuridine pulse labeling in healthy rhesus macaques ([i]Macaca mulatta[/i]) followed by hematology and flow cytometry analyses. Results were applied to a mathematical model, and the blood circulating half-life and daily production, respectively, of each cell type from macaques aged 5-10 y old were calculated for neutrophils (1.63 ± 0.16 d, 1.42 × 10 cells/l/d), basophils (1.78 ± 0.30 d, 5.89 × 10 cells/l/d), and CD14 CD16 classical monocytes (1.01 ± 0.15 d, 3.09 × 10 cells/l/d). Classical monocytes were released into the blood circulation as early as 1 d after dividing, whereas neutrophils remained in bone marrow 4-5 d before being released. Among granulocytes, neutrophils and basophils exhibited distinct kinetics in bone marrow maturation time and blood circulation. With increasing chronological age, there was a significant decrease in daily production of neutrophils and basophils, but the half-life of these granulocytes remained unchanged between 3 and 19 y of age. In contrast, daily production of classical monocytes remained stable through 19 y of age but exhibited a significant decline in half-life. These results demonstrated relatively short half-lives and continuous replenishment of neutrophils, basophils, and classical monocytes during homeostasis in adult rhesus macaques with compensations observed during increasing chronological age.
MeSH Terms
- Aging
- Animals
- Basophils
- Bone Marrow Cells
- Eosinophils
- Half-Life
- Homeostasis
- Macaca mulatta
- Male
- Monocytes
- Myeloid Cells
- Neutrophils
Children are more susceptible to Giardia lamblia infection. Cells and hormones contained in human colostrum have an immunoprotective action against giardiasis, but the effects of advanced maternal age on these components are poorly understood. This study analyzed the colostrum of older women to determine melatonin and cortisol levels besides the participation of these hormones on the functional activity of phagocytes against G. lamblia. Colostrum samples were collected from younger (18 to 35 years old) and older (over 36 years old) lactating women. Colostrum samples were subjected to melatonin and cortisol determination, immunophenotyping, quantification of superoxide release, and assessment of phagocytic rate and microbicidal activity of phagocytes treated with hormones and in the presence of G. lamblia. Colostrum from mothers of advanced age contained higher melatonin and cortisol levels and a lower rate of cells expressing CD14 and CD15 . In the colostru of these older mothers, melatonin increased superoxide release by phagocytes. In both groups, superoxide release by phagocytes treated with cortisol was higher in the presence of G. lamblia. In colostrum from mothers of advanced age, mononuclear (MN) phagocytes treated with melatonin showed higher phagocytosis of G. lamblia and higher microbicidal index. In younger mothers, MN and polymorphonuclear (PMN) colostrum phagocytes exhibited higher rates of G. lamblia elimination when treated with both melatonin and cortisol. In older mothers, cortisol and melatonin regulation for the functional activity of colostrum phagocytes against G. lamblia may represent an additional defense mechanism, relevant for the protection and treatment of parasitic infections in breastfed children.
MeSH Terms
- Adolescent
- Adult
- Aging
- Animals
- Child
- Colostrum
- Cross-Sectional Studies
- Female
- Giardia lamblia
- Giardiasis
- Humans
- Hydrocortisone
- Lactation
- Lewis X Antigen
- Lipopolysaccharide Receptors
- Maternal Age
- Melatonin
- Neutrophils
- Phagocytes
- Phagocytosis
- Pregnancy
- Superoxides
- Young Adult
Keywords
- Climacteric
- Colostrum
- Giardia lamblia
- Hormones
- Phagocytes
Human primary monocytes comprise a heterogeneous population that can be classified into three subsets based on CD14 and CD16 expression: classical (CD14 /CD16 ), intermediate (CD14 /CD16 ), and non-classical (CD14 /CD16 ). The non-classical monocytes are the most pro-inflammatory in response to TLR stimulation in vitro, yet they express a remarkably high basal level of miR-146a, a microRNA known to negatively regulate the TLR pathway. This concurrence of a pro-inflammatory status and a high miR-146a level has been associated with cellular senescence in other cell types. Hence, we assessed the three monocyte subsets for evidence of senescence, including proliferative status, telomere length, cellular ROS levels, and mitochondrial membrane potential. Indeed, the non-classical subset exhibited the clearest hallmarks of senescence, followed by the intermediate and then the classical subset. In addition, the non-classical subset secreted pro-inflammatory cytokines basally in vitro. The highly pro-inflammatory nature of the non-classical monocytes could be a manifestation of the senescence-associated secretory phenotype (SASP), likely induced by a high basal NF-κB activity and IL-1α production. Finally, we observed an accumulation of the non-classical monocytes, in conjunction with higher levels of plasma TNF-α and IL-8, in the elderly. These factors may contribute to inflamm-aging and age-related inflammatory conditions, such as atherosclerosis and osteoarthritis. With our new understanding that the non-classical monocyte subset is a senescent population, we can now re-examine the role of this subset in disease conditions where this subset expands.
MeSH Terms
- Adult
- Age Factors
- Aging
- Cell Proliferation
- Cells, Cultured
- Cellular Senescence
- Cytokines
- Female
- GPI-Linked Proteins
- Humans
- Inflammation
- Inflammation Mediators
- Interleukin-1alpha
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Male
- Membrane Potential, Mitochondrial
- MicroRNAs
- Middle Aged
- Mitochondria
- Monocytes
- NF-kappa B
- Phenotype
- Reactive Oxygen Species
- Receptors, IgG
- Signal Transduction
- Telomere Homeostasis
- Young Adult
Damage to the central nervous system during HIV infection can lead to variable neurobehavioral dysfunction termed HIV-associated neurocognitive disorders (HAND). There is no clear consensus regarding the neuropathological or cellular basis of HAND. We sought to study the potential contribution of aging to the pathogenesis of HAND. Aged (range = 14.7-24.8 year) rhesus macaques of Chinese origin (RM-Ch) (n = 23) were trained to perform cognitive tasks. Macaques were then divided into four groups to assess the impact of SIVmac251 infection (n = 12) and combined antiretroviral therapy (CART) (5 infected; 5 mock-infected) on the execution of these tasks. Aged SIV-infected RM-Ch demonstrated significant plasma viremia and modest CSF viral loads but showed few clinical signs, no elevations of systemic temperature, and no changes in activity levels, platelet counts or weight. Concentrations of biomarkers of acute and chronic inflammation such as soluble CD14, CXCL10, IL-6 and TNF-α are known to be elevated following SIV infection of young adult macaques of several species, but concentrations of these biomarkers did not shift after SIV infection in aged RM-Ch and remained similar to mock-infected macaques. Neither acute nor chronic SIV infection or CART had a significant impact on accuracy, speed or percent completion in a sensorimotor test. Viremia in the absence of a chronic elevated inflammatory response seen in some aged RM-Ch is reminiscent of SIV infection in natural disease resistant hosts. The absence of cognitive impairment during SIV infection in aged RM-Ch might be in part attributed to diminishment of some facets of the immunological response. Additional study encompassing species and age differences is necessary to substantiate this hypothesis.
MeSH Terms
- Age Factors
- Aging
- Animals
- Antibodies, Viral
- Antiretroviral Therapy, Highly Active
- Asymptomatic Diseases
- Brain
- CD4-Positive T-Lymphocytes
- CD8-Positive T-Lymphocytes
- Cognitive Dysfunction
- Disease Models, Animal
- Female
- HIV Infections
- Humans
- Macaca mulatta
- RNA, Viral
- Simian Immunodeficiency Virus
- Viral Load
- Viremia
Keywords
- Aging
- Cognition
- HIV
- HIV-associated neurocognitive disorder
- Rhesus macaque
- Simian immunodeficiency virus
Resveratrol generated enormous interest as it improved functions of multiple organs and could delay aging in animal models. However, basic mechanism of action was not understood and due to poor bioavailability, it has failed to enter the market. A highly active nano-formulation of resveratrol (XAR™) with enhanced bioavailability is now available. Present study was undertaken to evaluate its effects on stem cells biology in the human peripheral blood. Twelve healthy participants were enrolled of which five received XAR™, five were age-matched placebo controls and two were 76 and 85 years old. Peripheral blood was processed to study serum profile to monitor cardiac and pancreatic functions and subjected to density gradient centrifugation to enrich pluripotent (VSELs) and adult stem cells that get enriched along with red blood cells and in the Buffy coat respectively on Day 2 and Day 15 after XAR™ treatment. The XAR™ treatment resulted in an increased expression of pluripotency transcripts specific for VSELs (Oct-4A, Nanog and Sox2) on D2; specific transcripts for differentiation in the progenitors including Oct-4, Ikaros, CD14, CD90 on D15, and anti-ageing and tumor suppressor transcripts NAD, SIRT1, SIRT6 and p53 in both stem cells and progenitors. An improvement of cardiac and pancreatic markers in serum profile was also observed on D15. The decline in VSELs numbers with age and beneficial effects of the XAR™ treatment were evident by up-regulation of specific transcripts and on serum profile. XAR™ is a promising molecule that has the potential to activate pluripotent VSELs and tissue committed adult stem cells 'progenitors' resulting in the rejuvenation of various body tissues and for improved, cancer-free health with advanced age.
MeSH Terms
- Adult
- Adult Stem Cells
- Blood Buffy Coat
- Female
- Humans
- Male
- Middle Aged
- Nanog Homeobox Protein
- Octamer Transcription Factor-3
- Proliferating Cell Nuclear Antigen
- Resveratrol
- SOXB1 Transcription Factors
- Sirtuin 1
- Sirtuins
- Stem Cells
Keywords
- Aging
- OCT-4
- Regeneration
- Resveratrol
- Stem cells
- Tumor suppression
- VSELs
Aging is associated with a vasoconstrictive, pro-coagulant, and pro-inflammatory profile of arteries and a decline in the bioavailability of the endothelium-derived molecule nitric oxide. Dietary nitrate elicits vasodilatory, anti-coagulant and anti-inflammatory effects in younger individuals, but little is known about whether these benefits are evident in older adults. We investigated the effects of 140 mL of nitrate-rich (HI-NI; containing 12.9 mmol nitrate) versus nitrate-depleted beetroot juice (LO-NI; containing ≤0.04 mmol nitrate) on blood pressure, blood coagulation, vascular inflammation markers, plasma nitrate and nitrite before, and 3 h and 6 h after ingestion in healthy older adults (five males, seven females, mean age: 64 years, age range: 57-71 years) in a randomized, placebo-controlled, crossover study. Plasma nitrate and nitrite increased 3 and 6 h after HI-NI ingestion ([i]p[/i] < 0.05). Systolic, diastolic and mean arterial blood pressure decreased 3 h relative to baseline after HI-NI ingestion only ([i]p[/i] < 0.05). The number of blood monocyte-platelet aggregates decreased 3 h after HI-NI intake ([i]p[/i] < 0.05), indicating reduced platelet activation. The number of blood CD11b-expressing granulocytes decreased 3 h following HI-NI beetroot juice intake ([i]p[/i] < 0.05), suggesting a shift toward an anti-adhesive granulocyte phenotype. Numbers of blood CD14 CD16⁺ intermediate monocyte subtypes slightly increased 6 h after HI-NI beetroot juice ingestion ([i]p[/i] < 0.05), but the clinical implications of this response are currently unclear. These findings provide new evidence for the acute effects of nitrate-rich beetroot juice on circulating immune cells and platelets. Further long-term research is warranted to determine if these effects reduce the risk of developing hypertension and vascular inflammation with aging.
MeSH Terms
- Aged
- Aging
- Beta vulgaris
- Biomarkers
- Blood Platelets
- Blood Pressure
- CD11b Antigen
- Cardiovascular Diseases
- Cross-Over Studies
- Diet
- Double-Blind Method
- Female
- Fruit and Vegetable Juices
- Granulocytes
- Hemostasis
- Humans
- Inflammation
- Male
- Middle Aged
- Monocytes
- Nitrates
- Nitrites
- P-Selectin
- Plant Roots
- Prothrombin
- Thromboplastin
- Waist Circumference
Keywords
- aging
- anti-adhesive effects
- anti-thrombotic effects
- beetroot juice
- blood pressure
- dietary nitrate
- low-grade inflammation
- preserving vascular health
- thrombosis
Follicular fluid (FF) accumulates in the antrum of the ovarian follicle. In addition, FF provides the microenvironment for oocyte development, oocyte maturation and competence, which are acquired during follicular development. Superstimulatory treatment of 1-month-old lambs can achieve synchronous development of numerous growing follicles. However, these growing follicles are unable to completely mature and ovulate. Furthermore, the oocytes exhibit lower competence compared with those of ewes. In this study, we utilized an isobaric tag for relative and absolute quantification (iTRAQ)-based proteomics analysis and compare protein composition between pre-pubertal and adult superstimulated follicle FF in sheep. In total, 243 differentially expressed proteins were identified, including 155 downregulated and 88 upregulated between lamb and ewe. Gene ontology (GO) and KEGG pathway analysis indicated that the differentially expressed proteins are involved in signal transduction, anatomical structure development, stress response, metabolic pathways, and the complement and coagulation cascades. Many of the proteins known to affect follicle development were observed in lower abundance in FF of lamb (e.g. ADAMTS9, CD14, CTNNB1, FST, GCLC, HSPG2, IGFBP2, IGFBP6, INHBA, PRL, PAPPA, POSTN, PRDX1, SERPINA1, SOD3, STC1, VEGFC, etc.). However, a higher abundance was observed for proteasome proteins. Inadequate amounts of these proteins in FF may be lead to the unique characteristics of follicular development in lamb. These differentially expressed proteins illuminate the age-dependent changes in protein expression in the follicle microenvironment.
MeSH Terms
- Aging
- Animals
- Female
- Follicle Stimulating Hormone
- Follicular Fluid
- Gene Ontology
- Ovarian Follicle
- Proteomics
- Sexual Maturation
- Sheep, Domestic
Keywords
- follicular fluid
- lamb
- proteome
- quantitative proteomic
- superstimulation
Human cytomegalovirus (HCMV) primary infection and periodic reactivation of latent virus is generally well controlled by T-cell responses in healthy people. In older donors, overt HCMV disease is not generally seen despite the association of HCMV infection with increased risk of mortality. However, increases in HCMV DNA in urine of older people suggest that, although the immune response retains functionality, immunomodulation of the immune response due to lifelong viral carriage may alter its efficacy. Viral transcription is limited during latency to a handful of viral genes and there is both an IFNγ and cellular IL-10 CD4 T-cell response to HCMV latency-associated proteins. Production of cIL-10 by HCMV-specific CD4 T-cells is a candidate for aging-related immunomodulation. To address whether long-term carriage of HCMV changes the balance of cIL-10 and IFNγ-secreting T-cell populations, we recruited a large donor cohort aged 23-78 years and correlated T-cell responses to 11 HCMV proteins with age, HCMV IgG levels, latent HCMV load in CD14 monocytes, and T-cell numbers in the blood. IFNγ responses by CD4 and CD8 T-cells to all HCMV proteins were detected, with no age-related increase in this cohort. IL-10-secreting CD4 T cell responses were predominant to latency-associated proteins but did not increase with age. Quantification of HCMV genomes in CD14 monocytes, a known site of latent HCMV carriage, did not reveal any increase in viral genome copies in older donors. Importantly, there was a significant positive correlation between the latent viral genome copy number and the breadth and magnitude of the IFNγ T-cell response to HCMV proteins. This study suggests in healthy aged donors that HCMV-specific changes in the T cell compartment were not affected by age and were effective, as viremia was a very rare event. Evidence from studies of unwell aged has shown HCMV to be an important comorbidity factor, surveillance of latent HCMV load and low-level viremia in blood and body fluids, alongside typical immunological measures and assessment of the antiviral capacity of the HCMV-specific immune cell function would be informative in determining if antiviral treatment of HCMV replication in the old maybe beneficial.
Keywords
- IFNγ production
- cIL-10 CD4 T cells
- human cytomegalovirus
- human cytomegalovirus-specific T-cells
- immunology of aging
- latent viral load
- viral latency
Obesity prevalence among people living with HIV (HIV ) is rising. HIV and obesity are proinflammatory states, but their combined effect on inflammation (measured by interleukin 6, IL-6), altered coagulation (D-dimer), and monocyte activation (soluble CD14, sCD14) is unknown. We hypothesized inflammation increases when obesity and HIV infection co-occur. The Veterans Aging Cohort Study survey cohort is a prospective, observational study of predominantly male HIV veterans and veterans uninfected with HIV; a subset provided blood samples. Inclusion criteria for this analysis were body mass index ≥ 18.5 kg/m and biomarker measurement. Dependent variables were IL-6, sCD14, and D-dimer quartiles. Obesity/HIV status was the primary predictor. Unadjusted and adjusted logistic regression models were constructed. Data were analyzed for 1477 HIV and 823 uninfected participants. Unadjusted median IL-6 levels were significantly higher and sCD14 levels significantly lower in obese/HIV compared with nonobese/uninfected (P <0.01 for both). In adjusted analyses, the odds ratio for increased IL-6 in obese/HIV patients was 1.76 (95% confidence interval: 1.18 to 2.47) compared with nonobese/uninfected, and obesity/HIV remained associated with lower odds of elevated sCD14. We did not detect a synergistic association of co-occurring HIV and obesity on IL-6 or sCD14 elevation. D-dimer levels did not differ significantly between body mass index/HIV status groups. HIV-obesity comorbidity is associated with elevated IL-6, decreases in sCD14, and no significant difference in D-dimer. These findings are clinically significant, as previous studies associated these biomarkers with mortality. Future studies should assess whether other biomarkers show similar trends and potential mechanisms for unanticipated sCD14 and D-dimer findings.
MeSH Terms
- Adult
- Aging
- Biomarkers
- Comorbidity
- Cross-Sectional Studies
- Female
- Fibrin Fibrinogen Degradation Products
- HIV Infections
- Humans
- Inflammation
- Interleukin-6
- Lipopolysaccharide Receptors
- Male
- Middle Aged
- Obesity
- Prospective Studies
- United States
- Veterans
To evaluate the consequences of plasma from individuals with obesity on parameters associated with immunosenescence in unrelated healthy peripheral blood mononuclear cells (PBMC). Freshly isolated PBMC were incubated in media supplemented with 10% of plasma from individuals with obesity or control subjects for the first 4 hours of 24 to 120 hours of culture. Plasma from individuals with obesity modulated the phenotype of healthy PBMC, leading to a higher rate of apoptosis, lower amounts of phospho-γH2AX and -p53, and mitochondrial dysfunction. After 120 hours, there was a higher secretion of inflammatory cytokines IL-1β and IL-8. CD8 T lymphocytes presented decreased expression of CD28, which is associated with the immunosenescent phenotype. CD14 macrophages showed increased expression of CD80 and CD206, suggesting a modulation in the activation of macrophages. These results demonstrate that chronic systemic inflammation observed in obesity induces dysfunctional features in PBMC that are consistent with premature immunosenescence.
MeSH Terms
- Adult
- Apoptosis
- CD8-Positive T-Lymphocytes
- Culture Media
- Female
- Humans
- Immunosenescence
- Inflammation
- Interleukin-1beta
- Interleukin-8
- Leukocytes, Mononuclear
- Macrophages
- Male
- Obesity
- Serum
- Signal Transduction
β -Microglobulin (β M), the light chain of the major histocompatibility complex class I (MHC I), has been identified as a proaging factor and is involved in the pathogenesis of neurodegenerative disorders by driving cognitive and regenerative impairments. However, little attention has focused on the effect of β M in the development of lung emphysema. Here, we found that concentrations of β M in plasma were significantly elevated in patients with lung emphysema than those in normal control subjects (1.89 ± 0.12 vs. 1.42 ± 0.06 mg/l, [i]P[/i] < 0.01). Moreover, the expression of β M was significantly higher in lung tissue of emphysema (39.90 ± 1.97 vs. 23.94 ± 2.11%, [i]P[/i] < 0.01). Immunofluorescence showed that β M was mainly expressed in prosurfactant protein C-positive (pro-SPC ) alveolar epithelial cells and CD14 macrophages. Exposure to recombinant human β M and cigarette smoke extract (CSE) in vitro enhanced cellular senescence and inhibited proliferation of A549 cells, which was partially reversed by the presence of anti-β M antibody. However, anti-β M antibody did not attenuate the elevated production of IL-1β, IL-6, and TNF-α in A549 cells that were exposed to CSE. Immunofluorescence showed that colocalization of β M, and the hemochromatosis gene (HFE) protein was observed on A549 cells. These data suggest β M might participate in the development of lung emphysema through induction of lung epithelial cell senescence and inhibition.
MeSH Terms
- A549 Cells
- Antibodies
- Cell Proliferation
- Cellular Senescence
- Demography
- Epithelial Cells
- Female
- Humans
- Lung
- Male
- Middle Aged
- Phenotype
- Pulmonary Emphysema
- Smoking
- beta 2-Microglobulin
Keywords
- CSE
- emphysema
- epithelial cells
- senescence
- β2-microglobulin
Despite sepsis being a life-threatening disease, targeted drugs that improve the therapy of affected patients are still lacking. Infants and adults differ in the maturity level of their immune system and this results in distinct reactions to Gram-negative bacteria. To study reactions of human immune cells in vivo, we used NOD scid gamma mice transplanted with human CD34 stem cells to engraft a functional human immune system. Human cells undergo differentiation and maturation in these mice after transplantation and, accordingly, animals were divided into two groups: 8-13 wk and 15-22 wk after transplantation. Endotoxemia was induced by injecting LPS. Six h later, mice were euthanized. In both groups, LPS stimulation induced a decrease of CD14 monocytes in peripheral blood, an up-regulation of activation markers on different cell subsets such as myeloid dendritic cells, and a release of the human cytokines TNF-α, IL-6 and IL-10. However, significant differences were detected with regard to the amounts of released cytokines, and 8-13-wk-old mice produced more IL-6, while PTX3 was mainly released by 15-22-wk-old animals. Thus, here we provide a potential model for preclinical research of sepsis in infants and adults.
MeSH Terms
- Adult
- Aging
- Animals
- Antigens, CD34
- C-Reactive Protein
- Dendritic Cells
- Gram-Negative Bacterial Infections
- Humans
- Immune System
- Immunity, Cellular
- Infant
- Infant, Newborn
- Interleukin-6
- Lipopolysaccharides
- Mesenchymal Stem Cell Transplantation
- Mesenchymal Stem Cells
- Mice
- Mice, Inbred NOD
- Mice, SCID
- Sepsis
- Serum Amyloid P-Component
Keywords
- Humanized mice
- PTX3
- sepsis
- sepsis mouse model
The maintenance of chronic immune activation due to leishmaniasis or even due to microbial translocation is associated with immunosenescence and may contribute to frequent relapses. Our aim was to investigate whether patients with HIV-associated visceral leishmaniasis (VL/HIV) who experience a single episode of VL have different immunological behaviors in comparison to those who experience frequent relapses. VL/HIV patients were allocated to non-relapsing (NR, n = 6) and relapsing (R, n = 11) groups and were followed from the active phase of VL up to 12 months post-treatment (mpt). The patients were receiving highly active antiretroviral therapy (HAART) and secondary prophylaxis after VL therapy. During active VL, the two groups were similar in all immunological parameters, including the parasite load. At 6 and 12 mpt, the NR group showed a significant gain of CD4 T cells, a reduction of lymphocyte activation, and lower soluble CD14 and anti-Leishmania IgG3 levels compared to the R group. The viral load remained low, without correlation with the activation. The two groups showed elevated but similar percentages of senescent T cells. These findings suggest a decreased ability of the R group to downmodulate immune activation compared to the NR group. Such functional impairment of the effector response may be a useful indicator for predicting clinical prognosis and recommending starting or stopping secondary prophylaxis.
MeSH Terms
- Antibodies, Protozoan
- Bacterial Translocation
- CD4 Lymphocyte Count
- Coinfection
- Disease Progression
- HIV Infections
- Humans
- Immunity
- Immunoglobulin G
- Immunosenescence
- Leishmaniasis, Visceral
- Lymphocyte Activation
- Parasite Load
- Recurrence
- T-Lymphocyte Subsets
- Viral Load
Accumulation of tissue macrophages is a significant characteristic of disease-associated chronic inflammation, and facilitates the progression of disease pathology. However, the functional roles of these bone marrow-derived macrophages (BMDMs) in aging are unclear. Here, we identified agedependent macrophage accumulation in the bone marrow, showing that aging significantly increases the number of M1 macrophages and impairs polarization of BMDMs. We found that age-related dysregulation of BMDMs is associated with abnormal overexpression of the anti-inflammatory interleukin-10. BMDM dysregulation in aging impairs the expression levels of pro-inflammatory cytokines and genes involved in B-cell maturation and activation. Phagocytosis of apoptotic Jurkat cells by BMDMs was reduced because of low expression of phagocytic receptor CD14, indicating that increased apoptotic cells may result from defective phagocytosis of apoptotic cells in the BM of aged mice. Therefore, CD14 may represent a promising target for preventing BMDM dysregulation, and macrophage accumulation may provide diagnostic and therapeutic clues. [BMB Reports 2017; 50(1): 43-48].
MeSH Terms
- Aging
- Animals
- Anti-Inflammatory Agents
- Apoptosis
- Bone Marrow Cells
- Cell Differentiation
- Cytokines
- Humans
- Inflammation
- Interleukin-10
- Jurkat Cells
- Lipopolysaccharide Receptors
- Macrophages
- Male
- Mice
- Mice, Inbred C57BL
- Phagocytosis
HIV infection and biomarkers of inflammation [measured by interleukin-6 (IL-6)], monocyte activation [soluble CD14 (sCD14)], and coagulation (D-dimer) are associated with morbidity and mortality. We hypothesized that these immunologic processes mediate (explain) some of the excess risk of mortality among HIV infected (HIV ) versus uninfected people independently of comorbid diseases. Among 2350 (1521 HIV ) participants from the Veterans Aging Cohort Study Biomarker Cohort (VACS BC), we investigated whether the association between HIV and mortality was altered by adjustment for IL-6, sCD14, and D-dimer, accounting for confounders. Participants were followed from date of blood draw for biomarker assays (baseline) until death or July 25, 2013. Analyses included ordered logistic regression and Cox Proportional Hazards regression. During 6.9 years (median), 414 deaths occurred. The proportional odds of being in a higher quartile of IL-6, sCD14, or D-dimer were 2-3 fold higher for viremic HIV versus uninfected people. Mortality rates were higher among HIV compared with uninfected people [incidence rate ratio (95% CI): 1.31 (1.06 to 1.62)]. Mortality risk increased with increasing quartiles of IL-6, sCD14, and D-dimer regardless of HIV status. Adjustment for IL-6, sCD14, and D-dimer partially attenuated mortality risk among HIV people with unsuppressed viremia (HIV-1 RNA ≥10,000 copies per milliliter) compared with uninfected people-hazard ratio (95% CI) decreased from 2.18 (1.60 to 2.99) to 2.00 (1.45 to 2.76). HIV infection is associated with elevated IL-6, sCD14, and D-dimer, which are in turn associated with mortality. Baseline measures of these biomarkers partially mediate excess mortality risk among HIV versus uninfected people.
MeSH Terms
- Adult
- Aging
- Analysis of Variance
- Biomarkers
- Blood Coagulation Disorders
- Female
- Fibrin Fibrinogen Degradation Products
- HIV Infections
- HIV-1
- Humans
- Inflammation
- Interleukin-6
- Lipopolysaccharide Receptors
- Logistic Models
- Longitudinal Studies
- Male
- Middle Aged
- Monocytes
- United States
- Veterans
Rats of Albino Oxford (AO) strain in our animal facility exhibit a longer average healthy life span than rats of Dark Agouit (DA) strain. Since chronic activation of macrophages contributes to chronic low level inflammation common in older age, elucidation of the changes in middle-aged rats could be useful in prevention of unbalanced inflammatory response in advanced age. We have analysed the phenotype of unelicited and thioglycollate-elicited peritoneal macrophages from young and middle-aged DA and AO rats and tested functions of these cells following stimulation with lipopolysaccharide (LPS) in vitro. Unelicited cells from middle-aged DA rats produced higher amounts of proinflammatory mediators interleukin-6 (IL-6) and nitric oxide (NO), but have a diminished response to LPS stimulation then cells from young rats, in spite of increased frequency of TLR4- and CD14-expressing mature macrophages. Injection of thioglycollate robustly increased overall cytokine production in young rats' macrophages, while diminishing their response to LPS stimulation. In middle-aged DA rats injection of thioglycollate diminished IL-6 production, but increased it in response to LPS stimulation. Quite the contrary to DA rats, the macrophages from middle-aged AO rats have released diminished levels of TNF-α and NO, whereas urea production was strongly increased, when compared to the macrophages from young rats. Although the thioglycollate injection has increased the proportion of CD86 MHCII mature macrophages in young rats, and percentages of activated TLR4 macrophages in both age groups of AO rats, it has not affected the cytokine production in young rats' macrophages, and the TNF-α production in middle-aged rats' macrophages. Moreover, the injection of thioglycollate has robustly increased the production of urea in macrophages derived from both age groups of AO rats. Although middle-aged rats of both strains were healthy during experiment, differences between the inflammatory responses of peritoneal macrophages of middle-aged rats of these strains might be one of the contributing factors defining their health in their advanced age. Development of strategies for the prevention of undesirable inflammatory changes in the elderly would benefit from the prospective study of the middle-aged.
MeSH Terms
- Aging
- Animals
- Interleukin-6
- Lipopolysaccharides
- Macrophages, Peritoneal
- Male
- Nitric Oxide
- Peritonitis
- Rats
- Rats, Inbred Strains
- Thioglycolates
- Tumor Necrosis Factor-alpha
Keywords
- Macrophages
- Rat strains
- Thioglycollate-induced peritonitis
- Young and middle-aged rats
Alzheimer's disease (AD) is a neurodegenerative disease characterized by formation of amyloid-β (Aβ) plaques, activated microglia, and neuronal cell death leading to progressive dementia. Recent data indicate that microglia and monocyte-derived macrophages (MDM) are key players in the initiation and progression of AD, yet their respective roles remain to be clarified. As AD occurs mostly in the elderly and aging impairs myeloid functions, we addressed the inflammatory profile of microglia and MDM during aging in TgAPP/PS1 and TgAPP/PS1dE9, two transgenic AD mouse models, compared to WT littermates. We only found MDM infiltration in very aged mice. We determined that MDM highly expressed activation markers at basal state. In contrast, microglia exhibited an activated phenotype only with normal aging and Aβ pathology. Our study showed that CD14 and CD36, two receptors involved in phagocytosis, were upregulated during Aβ pathogenesis. Moreover, we observed, at the protein levels in AD models, higher production of pro-inflammatory mediators: IL-1β, p40, iNOS, CCL-3, CCL-4, and CXCL-1. Taken together, our data indicate that microglia and MDM display distinct phenotypes in AD models and highlight the specific effects of normal aging vs Aβ peptides on inflammatory processes that occur during the disease progression. These precise phenotypes of different subpopulations of myeloid cells in normal and pathologic conditions may allow the design of pertinent therapeutic strategy for AD.
MeSH Terms
- Aging
- Alzheimer Disease
- Amyloid beta-Peptides
- Animals
- CD36 Antigens
- Chemokines
- Disease Models, Animal
- Encephalomyelitis, Autoimmune, Experimental
- Inflammation
- Lipopolysaccharide Receptors
- Macrophages
- Mice, Inbred C57BL
- Mice, Transgenic
- Microglia
- Models, Biological
- Monocytes
- Myeloid Cells
- Nitric Oxide Synthase Type II
- Phagocytosis
- Phenotype
- Plaque, Amyloid
Keywords
- Alzheimer's disease
- aging
- chemokines
- cytokines
- macrophages
- microglia
Moderate consumption of red wine is associated with fewer cardiovascular events. We investigated whether red wine consumption counteracts the adverse vascular effects of cigarette smoking. Participants smoked 3 cigarettes alone or after drinking a titrated volume of red wine. Clinical chemistry, blood counts, plasma cytokine enzyme-linked immunosorbent assays, immunomagnetic separation of CD14 monocytes for gene expression analysis, fluorescence-activated cell sorting for microparticles, and isolation of circulating mononuclear cells to measure telomerase activity were performed, and urine cotinine levels were quantified. Compared with baseline, leukocytosis (P = .019), neutrophilia (P <.001), lymphopenia (P <.001), and eosinopenia (P = .008) were observed after only smoking. Endothelial and platelet-, monocyte-, and leukocyte-derived microparticles (P <.001 each) were elevated. In monocytes, messenger RNA expression of interleukin (IL)-6 (2.6- ± 0.57-fold), tumor necrosis factor alpha (2.2- ± 0.62-fold), and IL-1b (2.3- ± 0.44-fold) were upregulated, as was IL-6 (1.2 ± 0.12-fold) protein concentration in plasma. Smoking acutely inhibited mononuclear cell telomerase activity. Markers of endothelial damage, inflammation, and cellular aging were completely attenuated by red wine consumption. Cigarette smoke results in acute endothelial damage, vascular and systemic inflammation, and indicators of the cellular aging processes in otherwise healthy nonsmokers. Pretreatment with red wine was preventive. The findings underscore the magnitude of acute damage exerted by cigarette smoking in "occasional lifestyle smokers" and demonstrate the potential of red wine as a protective strategy to avert markers of vascular injury.
MeSH Terms
- Adult
- Cardiovascular System
- Cotinine
- Eosinophils
- Female
- Humans
- Interleukin-1beta
- Interleukin-6
- Leukocytosis
- Lymphopenia
- Male
- Neutrophils
- Smoking
- Telomerase
- Tumor Necrosis Factor-alpha
- Wine
Keywords
- Aging
- Endothelial function
- Inflammation
- Smoking
- Wine
Aging-associated noncommunicable comorbidities are more prevalent among human immunodeficiency virus type 1 (HIV)-infected individuals than among HIV-uninfected individuals. Residual HIV-related chronic immune activation and senescence may increase the risk of developing comorbidities. Immune phenotyping, thymic output, and telomere length were assessed in 94 HIV-infected individuals who were aged >45 years and receiving antiretroviral therapy (ART; cases) and 95 age-matched uninfected controls. Cases had lower CD4( ) T-cell counts, higher CD8( ) T-cell counts, and increased levels of immune activation (ie, increased soluble CD14 [sCD14] level and increased percentages of CD38( )HLA-DR( ) cells among both CD4( ) and CD8( ) T cells), regulatory T cells, and percentage of programmed cell death 1 (PD-1)-expressing cells among CD4( ) T cells. Immune senescence levels (ie, percentages of CD27(-)CD28(-) cells or CD57( ) cells) were comparable between cases and controls. Peripheral blood mononuclear cells from cases had shorter telomeres but increased single-joint T-cell receptor excision circle content and CD31( ) naive CD4( ) T cells. Although cytomegalovirus (CMV) antibody titers were higher in cases, CMV-specific T-cell responses were comparable between cases and controls. T-cell senescence in cases was independently associated with T-cell activation but not with CMV-specific immune responses. Despite long-term receipt of ART, HIV-infected adults had higher levels of immune activation, regulatory T cells, and PD-1-expressing CD4( ) cells and shorter telomeres. The increased soluble CD14 levels and percentage of CD38( )HLA-DR( ) cells among CD4( ) T cells correlated with shorter telomeres and increased regulatory T-cell levels. This suggests that HIV influences immune function irreversibly, with several pathways that are persistently abnormal during effective ART. Therapies aimed at improving immune health during ART are needed.
MeSH Terms
- Aged
- Aging
- Anti-Retroviral Agents
- Cohort Studies
- Female
- HIV Infections
- Humans
- Immunophenotyping
- Lipopolysaccharide Receptors
- Lymphocyte Activation
- Male
- Middle Aged
- Programmed Cell Death 1 Receptor
- T-Lymphocyte Subsets
- T-Lymphocytes, Regulatory
- Telomere
- Thymus Gland
Keywords
- ART
- HIV
- immune activation
- senescence
- telomeres
- thymic output
Cytomegalovirus (CMV) is a highly prevalent herpesvirus, which maintains lifelong latency and places a significant burden on host immunity. Infection is associated with increased rates of vascular disease and overall mortality in the elderly and there is an urgent need for improved understanding of the viral-host balance during ageing. CMV is extremely difficult to detect in healthy donors, however, using droplet digital PCR of DNA from peripheral blood monocytes, we obtained an absolute quantification of viral load in 44 healthy donors across a range of ages. Viral DNA was detected in 24 % (9/37) of donors below the age of 70 but was found in all individuals above this age. Furthermore, the mean CMV load was only 8.6 copies per 10,000 monocytes until approximately 70 years of age when it increased by almost 30 fold to 249 copies in older individuals (p < 0.0001). CMV was found within classical CD14 monocytes and was not detectable within the CD14-CD16 subset. The titre of CMV-specific IgG increased inexorably with age indicating that loss of humoral immunity is not a determinant of the increased viral load. In contrast, although cellular immunity to the structural late protein pp65 increased with age, the T cell response to the immediate early protein IE1 decreased in older donors. These data reveal that effective control of CMV is impaired during healthy ageing, most probably due to loss of cellular control of early viral reactivation. This information will be of value in guiding efforts to reduce CMV-associated health complications in the elderly.
Keywords
- Ageing
- Cytomegalovirus
- Lifespan
- Monocyte
- ddPCR
The aging of the human immunodeficiency virus type 1 (HIV-1)-infected population obligates a focus on the interaction between aging, comorbid conditions, and HIV-1. We recruited a cohort of HIV-1-infected men aged ≤ 35 years or ≥ 50 years who were receiving fully suppressive antiretroviral therapy (ART). We analyzed plasma markers of inflammation; T-cell activation, exhaustion, proliferation; and innate cellular subsets and functional capacity. Levels of lipopolysaccharide and the plasma marker of chemokine (C-C motif) ligand 2 were significantly elevated in older HIV-infected men despite comparable cellular phenotypes. Compared with similarly age-stratified uninfected subjects, older HIV-1-infected adults were also more frequently in the upper quartile of soluble CD14 expression.
MeSH Terms
- Adult
- Aging
- Anti-HIV Agents
- Bacterial Translocation
- Biomarkers
- Chemokine CCL2
- Genotype
- HIV Infections
- HIV-1
- Humans
- Immunity, Innate
- Inflammation
- Lymphocyte Activation
- Male
- Middle Aged
- T-Lymphocytes
Keywords
- HIV-1
- chemokine
- inflammation
- monocytes
Human immunodeficiency virus (HIV) infection and associated immune activation predict the risk of cardiovascular disease in resource-rich areas. Less is known about these relationships in sub-Saharan Africa. Beginning in 2005, we enrolled subjects in southwestern Uganda into a cohort at the time of antiretroviral therapy (ART) initiation. Multiple immune activation measures were assessed before and 6 months after ART initiation. Beginning in 2013, participants aged >40 years underwent metabolic profiling, including measurement of hemoglobin A1c and lipid levels and carotid ultrasonography. We fit regression models to identify traditional and HIV-specific correlates of common carotid intima media thickness (CCIMT). A total of 105 participants completed carotid ultrasonography, with a median completion time of 7 years following ART initiation. Age, low-density lipoprotein cholesterol level, and pre-ART HIV load were correlated with CCIMT. No association was found between CCIMT and any pre-ART biomarkers of immune activation. However, in multivariable models adjusted for cardiovascular disease risk factors, lower absolute levels of soluble CD14 and interleukin 6 and greater declines in the CD14 level and kynurenine-tryptophan ratio after 6 months of ART predicted a lower CCIMT years later (P < .01). Persistent immune activation despite ART-mediated viral suppression predicts the future atherosclerotic burden among HIV-infected Ugandans. Future work should focus on clinical correlates of these relationships, to elucidate the long-term health priorities for HIV-infected people in the region.
MeSH Terms
- Anti-HIV Agents
- Antigens, CD
- Biomarkers
- Carotid Artery Diseases
- Cohort Studies
- Cytokines
- Female
- Gene Expression Regulation
- HIV Infections
- Humans
- Male
- Middle Aged
- Risk Factors
- Uganda
Keywords
- HIV/AIDS
- Uganda
- aging
- antiretroviral therapy
- atherosclerosis
- carotid intima media thickness
- inflammation
Different immune alterations have been described in HIV-infected patients with visceral leishmaniasis (VL). We aimed to identify the immunological factors involved in the lack of immunological recovery and VL relapses in HIV-infected patients with VL, by comparison with other HIV-infected patients. We carried out a cross-sectional study of 55 patients receiving suppressive combination antiretroviral therapy (cART) for at least 1 year: nine with previous relapsing VL, 20 with an immunodiscordant response (IDR) to cART (CD4 count < 200 cells/μL) and no previous VL, and 26 with a concordant response (CR) to cART (CD4 count > 350 cells/μL) without VL. Immunosenescence was investigated by analysing CD57( ) CD28(-) levels, immune activation by analysing CD38( ) HLA-DR( ) levels, inflammation by analysing interleukin (IL)-6 levels, and microbial translocation by analysing lipopolysaccharide (LPS) and soluble CD14 (sCD14) levels. In VL patients, the median time since VL diagnosis was 42 months, and all patients had had at least one relapse despite suppressive cART for a median time of 43 months. Patients with previously diagnosed VL had a higher CD8 T-cell activation level (P < 0.001) than those with IDR. Also, levels of IL-6, LPS and especially sCD14, associated with bacterial translocation and additional monocyte activation, were significantly increased in patients with previous VL compared with patients with IDR (P = 0.048, P = 0.049 and P < 0.001, respectively). In addition, patients with previous VL had higher levels of CD8 T-cell senescence. Notably, the levels of immune activation and inflammation in patients with previous VL were not related to the time of VL diagnosis, the number of VL relapses, or hepatitis C virus (HCV) coinfection. Our data demonstrate that VL patients had an even worse immunological status than patients with IDR, which was probably associated with increased microbial translocation and additional monocyte/macrophage activation. These data explain the observed lack of immunological recovery and the occurrence of VL relapses in HIV-infected patients with previous VL.
MeSH Terms
- Adult
- Antiretroviral Therapy, Highly Active
- CD4 Lymphocyte Count
- Cellular Senescence
- Coinfection
- Cross-Sectional Studies
- Female
- Flow Cytometry
- Follow-Up Studies
- HIV Infections
- Humans
- Inflammation
- Leishmaniasis, Visceral
- Lymphocyte Activation
- Male
- Middle Aged
- Spain
- T-Lymphocytes
- Viral Load
Keywords
- HIV
- immune activation
- immunodiscordance
- immunosenescence
- leishmaniasis
Altered functions of macrophages with aging contribute to impairment of both innate and adaptive immunity in the elderly. The present study aimed to examine strain specificity of age-related changes in the phenotypic and functional characteristics of macrophages from DA and AO rats, which differ in average life span. Resident peritoneal macrophages from young (10-12 weeks old) and aged (98-104 weeks old) rats were tested for: (a) the surface expression of TLR4 and CD14; (b) the basal and LPS-induced production of TNF-α and IL-10; and (c) the basal and LPS-induced activity of iNOS and arginase, by measuring the levels of NO and urea, respectively, in the culture supernatants. Aging elevated TLR4 macrophage surface density in rats of both strains. Conversely, the age-related decrease in the surface density of CD14 co-receptor was detected only on macrophages from aged DA rats. Accordingly, with aging in DA rats, contrary to AO rats, upon LPS-stimulation both TNF-α and IL-10 levels decreased in culture supernatants. However, in rats of both strains TNF-α stimulation index (LPS-induced over basal production) remained stable with aging, but it was significantly greater in AO rats. Furthermore, with aging, IL-10 stimulation index decreased and increased in DA and AO rats, respectively. Age-related shift in urea stimulation index complied with the changes of IL-10 stimulation index during aging. In conclusion, the study suggests that the preserved ability of macrophages from aged AO rats to synthesize not only proinflammatory TNF-α, but also immunoregulatory IL-10 cytokine most likely contributes to their longer average life compared with DA rats.
MeSH Terms
- Adaptive Immunity
- Aging
- Animals
- Arginase
- Dipeptidyl Peptidase 4
- Female
- Immunity, Innate
- Interleukin-10
- Lipopolysaccharide Receptors
- Longevity
- Macrophages, Peritoneal
- Nitric Oxide
- Nitric Oxide Synthase Type II
- Rats
- Species Specificity
- Toll-Like Receptor 4
- Tumor Necrosis Factor-alpha
- Urea
A better understanding of the control of body fat distribution and muscle development is of the upmost importance for both human and animal physiology. This requires a better knowledge of the features and physiology of adult stem cells in adipose tissue and skeletal muscle. Thus the objective of the current study was to determine the type and proportion of these cells in growing and adult pigs. The different cell subsets of stromal vascular cells isolated from these tissues were characterized by flow cytometry using cell surface markers (CD11b, CD14, CD31, CD34, CD45, CD56, and CD90). Adipose and muscle cells were predominantly positive for the CD34, CD56, and CD90 markers. The proportion of positive cells changed with age especially in intermuscular adipose tissue and skeletal muscle where the percentage of CD90( ) cells markedly increased in adult animals. Further analysis using coimmunostaining indicates that eight populations with proportions ranging from 12 to 30% were identified in at least one tissue at 7 days of age, i.e., CD90( )/CD34( ), CD90( )/CD34(-), CD90( )/CD56( ), CD90( )/CD56(-), CD90(-)/CD56( ), CD56( )/CD34( ), CD56( )/CD34(-), and CD56(-)/CD34( ). Adipose tissues appeared to be a less heterogeneous tissue than skeletal muscle with two main populations (CD90( )/CD34(-) and CD90( )/CD56(-)) compared with five or more in muscle during the studied period. In culture, cells from adipose tissue and muscle differentiated into mature adipocytes in adipogenic medium. In myogenic conditions, only cells from muscle could form mature myofibers. Further studies are now needed to better understand the plasticity of those cell populations throughout life.
MeSH Terms
- Adipogenesis
- Adipose Tissue
- Adult Stem Cells
- Age Factors
- Aging
- Animals
- Biomarkers
- Cell Separation
- Cells, Cultured
- Female
- Flow Cytometry
- Immunohistochemistry
- Muscle Development
- Myoblasts, Skeletal
- Phenotype
- Swine
Keywords
- adipose tissue
- differentiation
- pig
- skeletal muscle
- stem cells
Aging is marked by a decline in physical function. Although the biological underpinnings for this remain unclear, loss of regenerative capacity has been proposed as one cause of the loss of physical function that occurs over time. The quantity of circulating progenitor cells (CPCs) may be one reflection of regenerative capability. We sought to determine whether certain specific CPC subpopulations were associated with physical function. Baseline CPCs were measured in 129 randomized participants in the Enhanced Fitness clinical trial based on the cell surface markers CD34, CD133, CD146, and CD14 and aldehyde dehydrogenase (ALDH) activity. Physical function was assessed using usual and rapid gait speed, 6-minute walk distance, chair stand time, and balance time. Low counts of early angiogenic CPCs identified as CD34( ), CD34( )CD133( ), and ALDH-bright (ALDH(br)) cells were associated with low usual gait speed (p < .005, p < .001, and p < .007), rapid gait speed (p < .001, p < .003, and p < .001), and 6-minute walking distance (all comparisons p < .001), and longer time required to complete five chair stands (p < .006, p < .002, and p < .004). CPC counts of mature endothelial or monocytic markers were not associated with physical function. The numbers of CD34( ) and ALDH(br) CPCs are significantly lower in patients with impaired physical function. Further studies are needed to determine the underlying causes for this association.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Aldehyde Dehydrogenase
- Antigens, CD
- Biomarkers
- Endothelial Cells
- Exercise Tolerance
- Geriatric Assessment
- Glucose Intolerance
- Humans
- Male
- Middle Aged
- Monocytes
- Physical Fitness
- Pilot Projects
- Statistics as Topic
- Stem Cells
Keywords
- Aging
- Endothelial progenitor cells
- Physical function
- Physical performance
- Progenitor cells biology.
The intricate interactions between the immune, endocrine and central nervous systems shape the innate immune response of the brain. We have previously shown that estradiol suppresses expression of immune genes in the frontal cortex of middle-aged ovariectomized rats, but not in young ones reflecting elevated expression of these genes in middle-aged, ovarian hormone deficient animals. Here, we explored the impact of menopause on the microglia phenotype capitalizing on the differential expression of macrophage-associated genes in quiescent and activated microglia. We selected twenty-three genes encoding phagocytic and recognition receptors expressed primarily in microglia, and eleven proinflammatory genes and followed their expression in the rat frontal cortex by real-time PCR. We used young, middle-aged and middle-aged ovariectomized rats to reveal age- and ovariectomy-related alterations. We analyzed the expression of the same set of genes in the postcentral and superior frontal gyrus of pre- and postmenopausal women using raw microarray data from our previous study. Ovariectomy caused up-regulation of four classic microglia reactivity marker genes including Cd11b, Cd18, Cd45 and Cd86. The change was reversible since estradiol attenuated transcriptional activation of the four marker genes. Expression of genes encoding phagocytic and toll-like receptors such as Cd11b, Cd18, C3, Cd32, Msr2 and Tlr4 increased, whereas scavenger receptor Cd36 decreased following ovariectomy. Ovarian hormone deprivation altered the expression of major components of estrogen and neuronal inhibitory signaling which are involved in the control of microglia reactivity. Strikingly similar changes took place in the postcentral and superior frontal gyrus of postmenopausal women. Based on the overlapping results of rat and human studies we propose that the microglia phenotype shifts from the resting toward the reactive state which can be characterized by up-regulation of CD11b, CD14, CD18, CD45, CD74, CD86, TLR4, down-regulation of CD36 and unchanged CD40 expression. As a result of this shift, microglial cells have lower threshold for subsequent activation in the forebrain of postmenopausal women.
MeSH Terms
- Adult
- Age Factors
- Aged
- Aging
- Animals
- Antigens, CD
- Cytokines
- Estradiol
- Estrogen Receptor alpha
- Female
- Frontal Lobe
- Gene Expression Regulation
- Histocompatibility Antigens
- Humans
- Menopause
- Middle Aged
- Ovariectomy
- Phagocytosis
- RNA, Messenger
- Rats
- Rats, Wistar
- Toll-Like Receptor 4
- Toll-Like Receptor 9
Physical function declines, and markers of inflammation increase with advancing age, even in healthy persons. Microbial translocation (MT) is the systemic exposure to mucosal surface microbes/microbial products without overt bacteremia and has been described in a number of pathologic conditions. We hypothesized that markers of MT, soluble CD14 (sCD14) and lipopolysaccharide (LPS) binding protein (LBP), may be a source of chronic inflammation in older persons and be associated with poorer physical function. We assessed cross-sectional relationships among two plasma biomarkers of MT (sCD14 and LBP), physical function (hand grip strength, short physical performance battery [SPPB], gait speed, walking distance, and disability questionnaire), and biomarkers of inflammation (C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-α), TNF-α soluble receptor 1 [[[TNF]]sR1]) in 59 older (60-89 years), healthy (no evidence of acute or chronic illness) men and women. LBP was inversely correlated with SPPB score and grip strength (p = .02 and p < .01, respectively) and positively correlated with CRP (p = 0.04) after adjusting for age, gender, and body mass index. sCD14 correlated with IL-6 (p = .01), TNF-α (p = .05), and TNFsR1 (p < .0001). Furthermore, the correlations between LBP and SPPB and grip strength remained significant after adjusting for each inflammatory biomarker. In healthy older individuals, LBP, a surrogate marker of MT, is associated with worse physical function and inflammation. Additional study is needed to determine whether MT is a marker for or a cause of inflammation and the associated functional impairments.
MeSH Terms
- Acute-Phase Proteins
- Aged
- Aged, 80 and over
- Aging
- Bacterial Translocation
- Biomarkers
- Carrier Proteins
- Cross-Sectional Studies
- Cytokines
- Exercise Test
- Female
- Gait
- Geriatric Assessment
- Hand Strength
- Humans
- Inflammation
- Inflammation Mediators
- Linear Models
- Male
- Membrane Glycoproteins
- Middle Aged
- Multivariate Analysis
- Physical Fitness
- Sensitivity and Specificity
{{medline-entry |title=ESeroS-GS modulates lipopolysaccharide-induced macrophage activation by impairing the assembly of TLR-4 complexes in lipid rafts. |pubmed-url=https://pubmed.ncbi.nlm.nih.gov/21276822 |abstract=The binding of lipopolysaccharides (LPS) to macrophages results in inflammatory responses. In extreme cases it can lead to endotoxic shock, often resulting in death. A broad range of antioxidants, including tocopherols, can reduce LPS activity in vitro and in vivo. To elucidate the underlying mechanisms of their action, we investigated the effect of the sodium salt of γ-L-glutamyl-S-[2-[[[3,4-dihydro-2,5,7,8-tetramethyl-2-(4,8,12-trimethyltridecyl)-2H-1-benzopyran-6-yl]oxy]carbonyl]-3-[[2-(1H-indol-3-yl)ethyl]amino]-3-oxopropyl]-L-cysteinylglycine (ESeroS-GS), a novel α-tocopherol derivative, on LPS-induced inflammation in vitro and in vivo. ESeroS-GS reduced the transcription of TNF-α, IL-1β, IL-6 and iNOS genes in a dose-dependent manner in RAW264.7 macrophages, and inhibited the release of these inflammatory factors. In addition, ESeroS-GS inhibited LPS-induced mortality in a mouse sepsis model. Electrophoretic mobility shift assays (EMSA) and reporter gene assays revealed that ESeroS-GS down-regulated the transcriptional activity of NF-κB. By analyzing the partitioning of CD14 and Toll-like receptor 4 (TLR-4) in cell membrane microdomains, we found that ESeroS-GS attenuates the binding of LPS to RAW264.7 cells via interfering with the relocation of CD14 and TLR-4 to lipid rafts, blocking the activation of interleukin-1 receptor-associated kinase 1 (IRAK-1), and inhibiting the consequent phosphorylation of TAK1 and IKKα/β, which together account for the suppression of NF-κB activation. Taken together, our data suggest that ESeroS-GS can modulate LPS signaling in macrophages by impairing TLR-4 complex assembly via a lipid raft dependent mechanism. This article is part of a Special Issue entitled: 11th European Symposium on Calcium. |mesh-terms=* Animals
- Benzopyrans
- Cell Line
- Cytokines
- Down-Regulation
- Fluorescein-5-isothiocyanate
- I-kappa B Kinase
- Indoles
- Inflammation Mediators
- Interleukin-1 Receptor-Associated Kinases
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Longevity
- MAP Kinase Kinase Kinases
- Macrophage Activation
- Macrophages
- Male
- Membrane Microdomains
- Mice
- Mice, Inbred C57BL
- Models, Biological
- Multiprotein Complexes
- NF-kappa B
- Nitric Oxide
- Nitric Oxide Synthase Type II
- Phosphorylation
- Sepsis
- Toll-Like Receptor 4
|full-text-url=https://sci-hub.do/10.1016/j.bbamcr.2011.01.019 }}
Young mice do not develop measurable periodontal bone loss, unless heavily infected with human periodontal pathogens. However, mice with a genetically altered immune system are unable to control their own oral flora and develop periodontitis early in life. Based on the potential of the indigenous oral microbiota to cause periodontitis, we hypothesized that normal mice may ultimately develop inflammatory periodontal bone loss, i.e. as a function of age. If confirmed, this could serve as an aging model of chronic periodontitis. Periodontal bone levels were measured as the distance from the cementoenamel junction to the alveolar bone crest in young mice (8-10 wk of age), old mice (>or= 18 mo of age) and mice of intermediate ages. Differential expression of inflammatory mediators in the gingivae of young and old mice was determined by quantitative real-time PCR. In comparison with young mice, old mice displayed significantly (p < 0.05) increased periodontal bone loss, accompanied by elevated expression of proinflammatory cytokines (interleukin-1 beta, tumor necrosis factor alpha and interleukin-17A) and innate immune receptors involved in the induction or amplification of inflammation (Toll-like receptor 2, CD14, CD11b, CD18, complement C5a receptor and triggering receptor expressed on myeloid cells 3). Mice develop naturally induced periodontal bone loss as a function of age. This aging model of periodontitis represents a genuinely chronic model to study mechanisms of periodontal tissue destruction.
MeSH Terms
- Aging
- Alveolar Bone Loss
- Alveolar Process
- Animals
- CD11b Antigen
- CD18 Antigens
- Chronic Periodontitis
- Disease Models, Animal
- Gingiva
- Immunity, Innate
- Inflammation Mediators
- Interleukin-17
- Interleukin-1beta
- Lipopolysaccharide Receptors
- Mice
- Mice, Inbred BALB C
- Mice, Inbred Strains
- Receptor, Anaphylatoxin C5a
- Receptors, Immunologic
- Toll-Like Receptor 2
- Tooth Cervix
- Tumor Necrosis Factor-alpha
Sepsis is a major cause of morbidity and mortality in the elderly population. In prior studies, we have shown that in vivo, the inflammatory response in aged animals is exaggerated as compared to young animals and that this response likely accounts for the increased morbidity and mortality. Part of this uncontrolled inflammatory response in sepsis is due to the innate immune response. However, recent studies have shown that the pathogenesis of sepsis is much more complex. The adrenergic autonomic nervous system is now thought to play a key role in modulating the inflammatory response in sepsis. In this study, we hypothesize that not only is the innate immune response enhanced in response to lipopolysaccharide (LPS) in aged animals, but that the adrenergic nervous system also plays a role in the release of excess inflammatory cytokines. Male Fischer-344 rats (young: 3 months; aged: 24 months) were used. Endotoxemia was induced by intravenous injection of lipopolysaccharide (LPS, 15 mg/kg BW). Splenic tissues were harvested and mRNA and protein were extracted. The protein expression of CD14 and TLR4, key mediators of LPS in the innate response, as well as alpha-2A adrenergic receptor (alpha(2A)-AR) and phosphodiesterase 4D (PDE4D), as the means by which the autonomic nervous system exerts its effects were analyzed. Splenic tissue concentrations of alpha(2A)-AR, PDE4D, CD14, and TLR4 were significantly increased in septic aged rats as compared to aged sham rats and septic young rats. The increased expression of alpha(2A)-AR in septic aged rats was further confirmed by immunohistochemical staining of splenic tissues. These data support the hypothesis that not only is the innate immune response increased in aged animals during sepsis, but that there is also an upregulated response of the adrenergic autonomic nervous system that contributes to excess proinflammatory cytokine release.
MeSH Terms
- Age Factors
- Aging
- Animals
- Cyclic Nucleotide Phosphodiesterases, Type 4
- Cytokines
- Endotoxemia
- Inflammation
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Male
- RNA, Messenger
- Rats
- Rats, Inbred F344
- Receptors, Adrenergic, alpha-2
- Sepsis
- Spleen
- Toll-Like Receptor 4
Natural killer (NK) cells are critical to the innate defence against intracellular infection. High NK cell frequencies have been detected in human neonates, which may compensate for the relative immaturity of the specific immune response. Additionally, phenotypic subsets of NK cells have been identified in humans with different functional properties. In this study, we examined the age distribution and phenotype of NK populations in bovine peripheral blood, including neonatal animals. We found that the NK cell populations defined by the phenotypes CD3(-)CD2( ) and NKp46( ) largely overlapped, so that the majority of NK cells in bovine peripheral blood were CD3(-)CD2( )NKp46( ). The remainder of the NK-like cells comprised two minor populations, CD3(-)CD2( )NKp46(-) and CD3(-)CD2(-)NKp46( ); the relative proportions of these varied with age. The lowest frequency of NK cells was recorded in 1-day-old calves, with the highest frequency in day 0 calves. The phenotypic characteristics of CD3(-)CD2( ) and NKp46( ) NK populations were similar; both populations expressed CD45RO, CD45RB, CD11b, CC84, CD8alphaalpha and CD8alphabeta and did not express CD21, WC1, CD14 or gammadelta TCR. Age-related phenotypic differences were apparent. The phenotypic characteristics of three NK subpopulations were described; a significantly greater proportion of the CD3(-)CD2(-)NKp46( ) population expressed CD8alpha compared to CD3(-)CD2( )NKp46( ) cells. Furthermore, a significantly greater proportion of the CD3(-)CD2( )NKp46(-) population expressed CD8 compared to total CD3(-)CD2( ) cells. Adult cattle had a significantly higher proportion of perforin( ) cells compared to calves aged </=6 weeks. In this age group, the majority of perforin( ) cells expressed NKp46, while in adults the majority of perforin( ) cells were NKp46(-). However, the proportion of NKp46( ) and CD3(-)CD2( ) cells that expressed perforin was not significantly different in any age group tested.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- CD2 Antigens
- CD3 Complex
- Cattle
- Humans
- Immunophenotyping
- Infant, Newborn
- Killer Cells, Natural
- Lymphocyte Count
- Lymphocyte Subsets
- Natural Cytotoxicity Triggering Receptor 1
- Perforin
- Species Specificity
Gout is the most common form of inflammatory arthritis in the elderly. In the last two decades, both hyperuricemia and gout have increased markedly and similar trends in the epidemiology of the metabolic syndrome have been observed. Recent studies provide new insights into the transporters that handle uric acid in the kidney as well as possible links between these transporters, hyperuricemia, and hypertension. The treatment of established hyperuricemia has also seen new developments. Febuxostat and PEG-uricase are two novel treatments that have been evaluated and shown to be highly effective in the management of hyperuricemia, thus enlarging the therapeutic options available to lower uric acid levels. Monosodium urate (MSU) crystals are potent inducers of inflammation. Within the joint, they trigger a local inflammatory reaction, neutrophil recruitment, and the production of pro-inflammatory cytokines as well as other inflammatory mediators. Experimentally, the uptake of MSU crystals by monocytes involves interactions with components of the innate immune system, namely Toll-like receptor (TLR)-2, TLR-4, and CD14. Intracellularly, MSU crystals activate multiple processes that lead to the formation of the NALP-3 (NACHT, LRR, and pyrin domain-containing-3) inflammasome complex that in turn processes pro-interleukin (IL)-1 to yield mature IL-1 beta, which is then secreted. The inflammatory effects of MSU are IL-1-dependent and can be blocked by IL-1 inhibitors. These advances in the understanding of hyperuricemia and gout provide new therapeutic targets for the future.
MeSH Terms
- Aging
- Animals
- Gout
- Humans
Flow cytometric analysis of human P2X(7) pore activity segregates variant from common P2RX7 genotypes and may serve as a biomarker for cancer, pain, inflammation, and immune responses to infection. Standardization is needed to accommodate variable sample age and instrumentation differences in a multicenter clinical trial. CD14-PE-stained whole blood samples were treated with YO-PRO-1 combined with a P2X(7) agonist (BzATP) or control, followed by the addition of PI after closure of the P2X(7) pore. Recalled instrument settings from previous publications were used to adapt a standardized fluorescent particle-adjusted set-up method. Experiments were performed to compare the two methods while evaluating components of systematic variability and facilitating reliable processing of samples with varied ages. The median YO-PRO-1 fluorescence of BzATP-treated samples had less variability when collected by the bead-adjusted method and was less influenced by the compensation strategy used. The average day-to-day coefficient of variance for assessments of P2X(7) pore activity by this method was 0.11 /- 0.04, and the exclusion of nonviable cells was found to accommodate samples aged up to 4 days after phlebotomy. The bead-adjusted set-up method produced measurements differing by only 2.0% /- 1.5% on two analog cytometers, and within similar decades when comparing analog to digital instruments. These results provide a standardized method for quantitative flow cytometric analysis of P2X(7) receptor phenotypes in blood monocytes with minimal intralaboratory variation and potential for interlaboratory comparisons that can greatly facilitate multicenter functional genomic clinical studies.
MeSH Terms
- Adenosine Triphosphate
- Aging
- Asthma
- Benzoxazoles
- Cell Survival
- Clinical Trials as Topic
- Flow Cytometry
- Fluorescence
- Humans
- Lipopolysaccharide Receptors
- Monocytes
- Multicenter Studies as Topic
- Phlebotomy
- Quinolinium Compounds
- Receptors, Purinergic P2
- Receptors, Purinergic P2X7
The aetiology of porcine post-weaning multisystemic wasting syndrome (PMWS) is poorly understood. Porcine circovirus type 2 (PCV-2) is an essential component of the experimental disease model for PMWS: however, evidence from experimental and field studies indicates that additional factors play a critical role in the aetiopathogenesis of PMWS. Current candidates include (1) immune stimulation (for example, via co-infection or vaccination), and (2) a novel infectious agent. A prospective, longitudinal case-control study was designed to investigate molecular triggers in leucocytes of neonatal piglets that may predispose to the development of PMWS. Blood samples were collected weekly from pigs (n=125) within five farms, from 1 week to 8 weeks of age: that is, before the appearance of clinical signs. Four colour flow cytometry was used to investigate changes in subsets of peripheral blood mononuclear cells, using monoclonal antibodies against the following cell associated markers; sIgG, CD3, MHCII dR, CD14, CD4a, CD8a, CD45RC, CD25, SWC3a, SWC8, CD163 and CD45. Sampling and laboratory analysis was supported by monitoring of clinical signs from 1 week to 20 weeks of age, or until disease supervened. At the conclusion of the study, 68 pigs (54%) were classified in Group 1 (no signs of clinical disease), 34 pigs (27%) in Group 2 (signs of clinical disease but not characteristic of PMWS), 17 pigs (14%) in Group 3 (suspect PMWS case) and 5 pigs (4%) in Group 4 (PMWS case). A single case of Porcine Dermatitis and Nephropathy syndrome (PDNS) was also diagnosed. Significant changes with age were demonstrated in clinically normal, neonatal pigs (Group 1), including an increase in B-cells and T-cells, and an increase in the proportion of total T-cells expressing MHCII. Within the T-cell subset, the proportion of CD8( high) CD4(-) T-cells increased, in addition to the proportion of CD4( ) T-cells co-expressing CD8. Of the factors recorded, farm was found to have a highly significant effect on immune system development in the neonate. Comparison of Groups 1 and 4 cases identified significant differences between pigs which remained normal and those which subsequently developed PMWS. Pigs which went on to develop PMWS had a greater proportion of T-cells expressing MHCII in early life, higher mean intensity of expression of MHCII on T-cells, higher mean intensity of expression of MHCII on B cells and higher expression of CD25 on CD45RC(-) T-cells. These findings suggest that lymphocyte activation may be a key early event in the aetiology of PMWS.
MeSH Terms
- Aging
- Animals
- Animals, Newborn
- Gene Expression Regulation
- Immunity, Cellular
- Major Histocompatibility Complex
- Porcine Postweaning Multisystemic Wasting Syndrome
- Swine
- T-Lymphocyte Subsets
Brain aging often results in cognitive impairment and is considered to be a major risk factor for neurodegenerative diseases. Earlier studies reported inflammatory responses in aged brain that could contribute to age-related neurodegeneration. Recently, innate immune receptors such as toll-like receptors (TLRs), so far implicated in defense against microorganisms, have been linked to pathogenesis of Alzheimer's disease. Therefore, we asked whether the transcription of TLRs (1-9) and CD14, could also be altered in physiological brain aging. Using real-time polymerase chain reaction (PCR), we indeed observed that TLR1, TLR2, TLR4, TLR5, TLR7 and CD14 expression was up-regulated in mouse brain in correlation with age. In contrast, transcriptions of TLR3, TLR6 and TLR8 were unchanged and the one of TLR9 was down-regulated. In situ hybridization further confirmed these results and identified the cellular source of TLR2 and TLR7 as mononuclear phagocytes. Together, this first systematic analysis demonstrates altered regulation of those innate immune receptors even in normal brain aging, which might be of relevance for understanding susceptibility to neurodegenerative processes associated with aging.
MeSH Terms
- Age Factors
- Aging
- Animals
- Brain
- Gene Expression Regulation
- In Situ Hybridization
- Lipopolysaccharide Receptors
- Mice
- Mice, Inbred C57BL
- RNA, Messenger
- Reverse Transcriptase Polymerase Chain Reaction
- Toll-Like Receptors
The impaired infection control related to the functional immaturity of the neonatal immune system is an important cause of infection in preterm newborns. We previously reported that constitutive Toll-like receptor (TLR) 4 expression and cytokine secretion on lipopolysaccharide (LPS) stimulation increases with gestational age. Here, we analyzed constitutive monocyte TLR2 expression and evaluated the expression profiles of the proximal downstream adapter molecule myeloid differentiation factor 88 (MyD88). We further investigated activation of protein kinases p38 and extracellular regulated kinsase (ERK) 1/2 in CD14 monocytes after ex vivo stimulation with bacterial TLR ligands (LPS and lipoteichoic acid [[[LTA]]]). The functional outcome of the stimulation was determined by cytokine secretion. Monocytes from 31 preterm newborns (<30 weeks of gestation, n=16; 30-37 weeks of gestation, n=15), 10 term newborns, and 12 adults were investigated. In contrast to TLR4 expression, TLR2 levels did not differ between age groups. However, MyD88 levels were significantly lower in preterm newborns. Activation of p38 and ERK1/2 was impaired in all newborn age groups after stimulation with TLR-specific ligands. Accordingly, after LTA stimulation, the levels of interleukin (IL)-1 beta , IL-6, and IL-8 cytokine production were substantially lower (P<.001) in preterm newborns than in adults. The reduced functional response to bacterial cell wall components appears to be part of the functional immaturity of the neonatal immune system and might predispose premature newborns to bacterial infection.
MeSH Terms
- Adult
- Aging
- Female
- Fetal Blood
- Gene Expression Regulation, Developmental
- Gestational Age
- Humans
- Immunity, Innate
- Infant, Newborn
- Infant, Premature
- Monocytes
- Myeloid Differentiation Factor 88
- Pregnancy
- Signal Transduction
- Toll-Like Receptor 2
- Toll-Like Receptor 4
- Toll-Like Receptors
The aim of this study was to investigate the distribution of immune cells in the endometrium of anoestrous female pigs, five sows in anoestrus by lactation and five pre-pubertal gilts (Swedish Landrace x Swedish Yorkshire). Uterine samples, taken immediately after slaughter, were fixed, embedded in plastic resin and stained with toluidine blue or cryo fixed and stored in a freezer at -70 degrees C until analysed by immunohistochemistry with an avidin-biotin peroxidase method. Immune cells in the surface (luminal) and the glandular epithelium as well as the subepithelial and the glandular connective tissue layers were counted using light microscopy. In the surface (luminal) and the glandular epithelia of gilts and sows, lymphocytes were the predominant immune cells found. There were no significant differences between gilts and sows. Macrophages were detected in the glandular epithelium of sows but not in gilts. In the subepithelial and the glandular connective tissue layers of both gilts and sows, lymphocytes were also the most common immune cells found. The numbers of lymphocytes and macrophages were significantly higher in the sows than in the gilts (p <or= 0.05) in both the layers of connective tissue. Numbers of plasma cells, mast cells, eosinophils and neutrophils in the connective tissue were low and not significantly different between sows and gilts. In both the surface (luminal) epithelium and the subepithelial connective tissue, higher numbers of CD2 than CD3 positive cells were found (p <or= 0.01). The numbers of CD2 positive cells in both epithelium and connective tissue and the number of CD3 positive cells in the epithelium were significantly higher in the sows than the gilts (p <or= 0.05). A few CD79 positive cells were found in the subepithelial connective tissue and none in the epithelia. A few CD14 and SWC3 positive cells were found in the epithelia. The numbers of CD14, SWC3 and MHC class II positive cells were significantly higher in the sows than in the gilts (p <or= 0.05) in the subepithelial connective tissue. In conclusion, the distribution of immune cells in the endometrium of anoestrous female pigs was affected by experienced pregnancy and parturition. In sows with lactation-induced anoestrus, there was a markedly higher cell infiltration (lymphocytes and macrophages) than in the pre-pubertal gilts. In pre-pubertal gilts, lymphocytes dominated, which indicates a role in the maturation of the endometrium.
MeSH Terms
- Aging
- Anestrus
- Animals
- Endometrium
- Female
- Immunohistochemistry
- Lactation
- Lymphocyte Subsets
- Lymphocytes
- Macrophages
- Sexual Maturation
- Swine
Dysfunctional inflammatory responses have been implicated in several neonatal inflammatory disorders following infection and hypoxia. We aimed to study the effects of in vitro hypoxia and heat shock (HS) on normal adult and newborn neutrophil migration (CD11b) and persistence (apoptosis) following lipopolysaccharide (LPS) stimulation. The mechanism for altered LPS responses was assessed at the level of the LPS signalling receptors, Toll-like receptor-4 (TLR-4), TLR-2 and CD14 expression in normal neonates and adults. In adults, although hypoxia delayed neutrophil apoptosis, LPS enhanced this response. In contrast, HS (42 degrees C) increased adult apoptotic rates and abrogated the LPS responses. Both hypoxia and HS prevented the LPS-induced increase in adult CD11b although it was unaltered in neonates. Adult TLR-4 neutrophil expression was increased by LPS and hypoxia, and decreased in HS, possibly explaining their variable LPS responsiveness. In contrast, neonatal neutrophils were LPS hyporesponsive which may be mediated by failure of TLR-4 upregulation with LPS. Neonates do not have increased LPS responsiveness in hypoxia or heat shock in vitro, which may prevent hyperinflammation and thereby minimise tissue damage in inflammation or infection.
MeSH Terms
- Adult
- Aging
- Apoptosis
- CD11b Antigen
- Cell Hypoxia
- Female
- Fetal Blood
- Gestational Age
- Hot Temperature
- Humans
- Infant, Newborn
- Lipopolysaccharides
- Male
- Neutrophils
- Toll-Like Receptor 4
A reduction in macrophage (MPhi) function with aging makes mice less responsive to bacterial capsular polysaccharides, such as those present in the pneumococcal polysaccharide vaccine, a model of thymus independent (TI) antigen (Ag). Using trinitrophenol (TNP)-lipopolysaccharide (LPS) and TNP-Ficoll, two other well-studied TI Ag, we studied the mechanistic basis of reduced MPhi function in the aged. We show that aged mice are profoundly hyporesponsive to these TI Ag. As a result of a requirement for MPhi, highly purified B cells from young-adult mice do not respond to TI Ag. When purified, young B cells were immunized with TNP-Ficoll, the antibody production from those cultures reconstituted with MPhi from aged mice was significantly lower than that seen with young MPhi. Consequently, this unresponsiveness can be overcome by a mixture of interleukin (IL)-1beta and IL-6. Upon stimulation with LPS, in comparison with young MPhi, aged MPhi secreted reduced amounts of IL-6, tumor necrosis factor alpha, IL-1beta, and IL-12, cytokines necessary for B cells to respond to TI Ag. LPS also induced aged MPhi to produce an excess of IL-10. Neutralization of IL-10 enhanced the production of proinflamatory cytokines by MPhi upon LPS stimulation and also induced Ab production by aged splenocytes. Thus, the inability of aged MPhi to help the B cell response appears to be caused by an excess of IL-10. As aged MPhi have a reduced number of cells expressing Toll-like receptor 4 and CD14, the imbalance in cytokine production might be partly a result of fewer cells expressing key components of the LPS receptor complex.
MeSH Terms
- Aging
- Animals
- B-Lymphocytes
- Ficoll
- Inflammation
- Interleukin-10
- Interleukin-6
- Lipopolysaccharides
- Lymphocytes
- Macrophages
- Mice
- Mice, Inbred BALB C
- Mice, Inbred C57BL
- Picrates
- Spleen
The protein CD14 is a pattern recognition receptor for bacterial lipopolysaccharide (LPS). Whether genetic variation has the same influence on soluble CD14 (sCD14) levels in human plasma and milk remains to be determined. We measured sCD14 levels in plasma during pregnancy (n = 196) and in milk in the postpartum (n = 152) for women genotyped for the single nucleotide polymorphisms (SNPs) at positions -1619, -550, and -159 from the transcription start site of the CD14 gene. Plasma- and milk-sCD14 levels differed significantly both by CD14/-1619 and CD14/-550 genotypes and by haplotypes. Most interestingly, sCD14 levels were regulated differentially by the same genetic variants in plasma and milk, with the CD14/-550T allele and the corresponding are ATC haplotype associated with high sCD14 in milk but low sCD14 in plasma. A correlation between sCD14 levels in plasma and milk was absent (r = 0.091, P = NS). Our findings suggest the existence of cell-specific regulation mechanisms of CD14 gene expression.
MeSH Terms
- Adult
- Aging
- Breast Feeding
- Ethnic Groups
- Female
- Gene Expression Regulation
- Genotype
- Humans
- Lipopolysaccharide Receptors
- Milk, Human
- Parity
- Polymorphism, Single Nucleotide
- Pregnancy
- Solubility
Aging is the result of several detrimental changes that lead to a decrease in homeostasis, an increase in the incidence of degenerative diseases, and death. A caloric-restricted diet (CR), which consists of a significant reduction in calorie intake (40%) without malnutrition, has been shown to delay the onset of age-related diseases and pathologies and to extend life span. The aims of this study were to assess the effects of aging and CR on lipopolysaccharide (LPS)-dependant cytokine production by peritoneal macrophages (PMphis). Resident naïve PMphis were isolated from 2- to 24-month-old male C57BL/6 mice and were stimulated with Escherichia coli LPS (100 ng/mL) for 1 to 5 h in culture conditions. A linear decrease in the production of LPS-induced tumor necrosis factor alpha (TNF-alpha) and interleukin (IL) 10 was observed with age. LPS-induced IL-6 and IL-1beta levels were also reduced with age, but in a nonlinear fashion. Expression of CD14, the major receptor for LPS, on the PMphi surface was also observed to decline with age. Moreover, TNF-alpha production by PMphis was reduced in mice undergoing the two different CR diets of limited daily feeding and intermittent fasting, as compared with ad libitum-fed mice. The results of this study add the new variables age and diet to the paradigm proposing that the response to LPS is modulated by multiple components, including genetic background and sex.
MeSH Terms
- Aging
- Animals
- Cells, Cultured
- Diet, Reducing
- Energy Intake
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Macrophages, Peritoneal
- Male
- Mice
- Mice, Inbred C57BL
Toll receptors in Drosophila contribute to host defense and establish the body plan. Mammalian homologues of Toll, the Toll-like receptors (TLRs), are thought to function only in host defense. Here, we report that mice harboring mutations in TLR4 or in CD14, a co-receptor for TLR4, have an "ideal" body plan consisting of increased bone mineral content, density, and size as well as decreased body fat. These mutant mice live long lives, have normal activity and fertility, and show no evidence of infection. Unlike many strains of caged wild-type mice, they do not become obese. Although all mice continue to gain body fat, bone content, and overall weight, the difference in bone content and body fat between mutant and wild-type mice increases with age. Thus, defects in TLR4/CD14 complex generate an "Adonis" phenotype, characterized by this ideal body type, and this function could potentially be exploited for the treatment of osteoporosis and obesity.
MeSH Terms
- Adipose Tissue
- Aging
- Animals
- Body Constitution
- Body Patterning
- Bone Density
- Bone and Bones
- Female
- Fertility
- Gene Deletion
- Lipopolysaccharide Receptors
- Longevity
- Male
- Mice
- Mice, Inbred BALB C
- Mice, Inbred C3H
- Mice, Inbred C57BL
- Mice, Mutant Strains
- Motor Activity
- Obesity
- Osteoporosis
- Phenotype
- Receptors, Cell Surface
- Sequence Deletion
- Thinness
- Toll-Like Receptor 4
- Weight-Bearing
A reduced microbial stimulation has been reported as a reason for the increasing prevalence of atopic diseases in industrialized countries. Antigen-presenting cells (APC), responding to microbial signals by pattern recognition receptors such as CD14, have an important role in the development of the Th1/Th2 balance. We hypothesized that atopic children have a lower expression of CD14 on monocytes and lower soluble CD14 levels (sCD14). Seventy-six children were followed prospectively from birth and signs of atopic disease were evaluated. The expression of CD14 on monocytes was analysed with flow cytometry at 0, 3, 6, 12 and 18 months. Circulating levels of sCD14 were analysed by ELISA and total IgE was analysed by fluoroenzymo immunoassay at these ages, and in a subgroup, followed up at 7 years. Levels of sCD14 were reduced at 7 years both in children with a current or a cumulative history of atopy compared to non-atopic children with P=0.002 and 0.001, respectively. Sensitized children with atopic symptoms had lower sCD14 at 3 and 18 months and at 7 years of age than non-atopic non-sensitized children with P=0.023, 0.039 and 0.008, respectively. The lower levels of sCD14 observed in atopic children may be a consequence of an atopic family heredity and/or atopic disease, but it may also reflect a reduced capacity to respond to microbial signals.
MeSH Terms
- Aging
- Antigen-Presenting Cells
- Follow-Up Studies
- Humans
- Hypersensitivity, Immediate
- Immunoglobulin E
- Infant, Newborn
- Lipopolysaccharide Receptors
- Monocytes
- Skin Tests
- Solubility
Exposure to furred pets might confer protection against the development of allergic sensitization through a mechanism that is incompletely understood. The objective of this study was to determine the effects of pet exposure and genotype on immunologic development and the incidence of atopic markers and diseases in the first year of life. Pet exposure in the home was compared with cytokine secretion patterns (mitogen-stimulated mononuclear cells at birth and age 1 year) and indicators of atopy (allergen-specific and total IgE, eosinophilia, food allergy, atopic dermatitis) in 285 infants. Interactions with genotype at the CD14 locus were also evaluated in the data analyses. Exposure to dogs was associated with reduced allergen sensitization (19% vs 33%, P =.020) and atopic dermatitis (30% vs 51%, P <.001). The risk for atopic dermatitis was further influenced by genotype at the CD14 locus (P =.006), even after adjusting for exposure to dogs (P =.003). Furthermore, infants with the genotype -159TT were less likely to develop atopic dermatitis if they were exposed to a dog (5% vs 43%, P =.04). Last, dog exposure was associated with increased IL-10 (117 vs 79 pg/mL, P =.002) and IL-13 (280 vs 226 pg/mL, P =.013) responses at age 1 year. Having a dog in infancy is associated with higher IL-10 and IL-13 cytokine secretion profiles and reduced allergic sensitization and atopic dermatitis. These findings suggest that postnatal exposure to dogs can influence immune development in a genotype-specific fashion and thereby attenuate the development of atopy in at-risk children.
MeSH Terms
- Adult
- Aging
- Allergens
- Animals
- Animals, Domestic
- Cats
- Cytokines
- Dermatitis, Atopic
- Dogs
- Food Hypersensitivity
- Genotype
- Humans
- Hypersensitivity, Immediate
- Infant
- Infant, Newborn
- Interleukin-10
- Interleukin-13
- Lipopolysaccharide Receptors
- Risk Factors
To investigate the effect of the functional CD14 promoter polymorphism on serum liver function tests and abnormally elevated liver function tests in a general population sample. We recruited 310 subjects at random from general practitioner lists in Surrey. A previously validated medical questionnaire was completed and a serum sample provided for estimation of liver function test and genotyping of CD14 promoter polymorphism. The TT polymorphism was associated with significantly reduced serum levels of alanine aminotransferase 0.49 (95% confidence interval 0.26-0.93), gamma-glutamyl transferase 0.49 (95% confidence interval 0.38-0.89) and aspartate aminotransferase 0.48 (95% confidence interval 10.38-0.89). The TT polymorphism was associated with a reduced frequency of liver function test abnormalities, especially gamma-glutamyl transferase (odds ratio 0.113; 0.015-0.851). The TT promoter polymorphism was associated with reduced serum levels of alanine aminotransferase, gamma-glutamyl transferase and aspartate aminotransferase in healthy patients, and a low level of liver function test abnormalities. The relationship with gamma-glutamyl transferase stands after correction for age, gender, obesity and alcohol consumption. This raises the possibility that this genotype may offer protection from the development of fatty liver disease.
MeSH Terms
- Aged
- Aging
- Alanine Transaminase
- Alcohol Drinking
- Aspartate Aminotransferases
- Body Mass Index
- Female
- Genetic Predisposition to Disease
- Genotype
- Humans
- Lipopolysaccharide Receptors
- Liver
- Liver Function Tests
- Male
- Middle Aged
- Obesity
- Polymorphism, Genetic
- Promoter Regions, Genetic
- gamma-Glutamyltransferase
To understand bone metabolism during senescence, we examined age-related change in nitric oxide (NO) production from bone marrow cells stimulated by lipopolysaccharide (LPS). We evaluated the age-related change in the NO production and expression of iNOS protein and mRNA of LPS-stimulated bone marrow cells collected from the tibiae of young and retired female and young and retired male rats. In addition, we used flow cytometry to assess changes in the distribution of CD14, a cell surface receptor of LPS. The results revealed that NO production from bone marrow cells stimulated with LPS changed with aging. The NO levels in old rats were significantly higher (P<0.05) than those in young rats. Polymerase chain reaction (PCR) analysis indicated that the LPS-induced expression of iNOS mRNA was augmented in retired rats. Although the distribution pattern of the bone marrow cells was similar between young and retired rats, the percentage of CD14-positive cells in specific populations differed between the age groups. Specifically, in the granule-containing bone marrow cells, the percentage of CD14-positive cells was increased in retired rats. Our results indicate that LPS-stimulated NO production from rat bone marrow cells increased with age and that the difference in responsiveness might be due to changes in the percentage of CD14-positive cells in the bone marrow.
MeSH Terms
- Aging
- Animals
- Bone Marrow Cells
- Bone Resorption
- Cells, Cultured
- DNA Primers
- Female
- Flow Cytometry
- Lipopolysaccharides
- Male
- Nitric Oxide
- Nitric Oxide Synthase
- Nitric Oxide Synthase Type II
- Polymerase Chain Reaction
- RNA, Messenger
- Rats
- Rats, Sprague-Dawley
The purpose of this study was to examine the influence of resistive exercise training and hormone status on mRNA expression of toll-like receptor 4 (TLR4), CD14, IL-1beta, IL-6, and TNF-alpha. Resistive exercise-trained women on "traditional" hormone replacements [hormone replacement therapy (HRT), n = 9], not taking hormones (NHR, n = 6), or taking medications known to influence bone (MIB, n = 7) were compared with untrained subjects not taking supplemental hormones (Con, n = 6). Blood was taken from trained subjects before, immediately after, and 2 h after resistive exercise (same time points for resting Con). TLR4 mRNA expression (RT-PCR) was not different among groups or across time but was significantly (P = 0.044) lower (1.9-fold) when trained groups were collapsed and compared with Con. There was also a significant group effect (P < 0.0001) for TLR4 mRNA when expressed per monocyte. CD14 expression was significantly (P = 0.006) lower (2.3-fold) for training groups collapsed and compared with Con. CD14 mRNA, expressed per monocyte, was significantly lower immediately after resistive exercise for NHR, HRT, and MIB compared with Con. There were few significant effects detected for IL-6, IL-1beta, and TNF-alpha mRNA, but there was a significant group effect (P < 0.0001) for TNF-alpha mRNA expressed per monocyte (Con > HRT, NHR, MIB). These findings suggest that there may be a resistive exercise training-induced reduction in TLR4/CD14 expression in older women. Further research is needed to determine whether lower TLR4/CD14 could explain the lower LPS-stimulated inflammatory cytokines observed in these women.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Cells, Cultured
- Estradiol
- Estrogen Replacement Therapy
- Exercise
- Female
- Gene Expression
- Humans
- Interleukin-1
- Interleukin-6
- Leukocyte Count
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Membrane Glycoproteins
- Monocytes
- Physical Fitness
- Postmenopause
- Progesterone
- RNA, Messenger
- Receptors, Cell Surface
- Toll-Like Receptor 4
- Toll-Like Receptors
- Tumor Necrosis Factor-alpha
Different types of circulating dendritic cells have been described. Dendritic cells influence differentiation of naive T lymphocytes into T helper type 1 (Th1) and Th2 effector cells. The purpose of this study was to evaluate the number of circulating DC subtypes in peripheral blood of allergic and healthy children and in cord blood of neonates from allergic and non-allergic parents. Circulating dendritic cells were flow cytometrically identified in whole blood samples as lineage (CD3, CD14, CD16, CD19, CD20, CD56) negative, CD34 negative and HLA-DR-positive cells. According to the expression of CD123 and CD11c, different DC subtypes were identified. Apart from DC1 (CD11c CD123dim ) and DC2 (CD11c- CD123high ), a third DC population was described with less differentiated phenotypic characteristics, namely CD11c- CD123dim , and therefore defined here as less differentiated DC (ldDC). These ldDC represented the major DC population in cord blood and showed an age-depended decrease. The highest level of ldDC was detected in children with atopic dermatitis, whereas asthmatic children showed the lowest ldDC counts. Furthermore, high-dose inhaled corticosteroid treatment in asthmatic children was related to a decreased ldDC number. The number of circulating DC2 was significantly lower in allergic children, especially in asthmatics, compared to healthy children. In cord blood, no differences in DC subtypes were detectable between neonates at low and high risk for allergic disorders. These results indicate that, apart from DC1 and DC2, a third population of dendritic cells, identified as CD11c- CD123dim cells and defined as less differentiated DC, must be considered in the evaluation of circulating DC. Furthermore, DC2 counts were decreased in allergic children, especially in asthmatics, which might be the consequence of an increased recruitment to the target organs.
MeSH Terms
- Adolescent
- Aging
- Asthma
- Blood Cell Count
- Cell Separation
- Child
- Child, Preschool
- Dendritic Cells
- Dermatitis, Atopic
- Female
- Fetal Blood
- Flow Cytometry
- Humans
- Hypersensitivity, Immediate
- Immunophenotyping
- Infant
- Infant, Newborn
- Male
- Maternal-Fetal Exchange
- Pregnancy
The T helper type-2 (Th2)-dominated situation can be observed in allergic diseases such as asthma or atopic dermatitis. A reduced ability to produce IL-12, which is a key cytokine for the induction of Th1 responses, has been proposed to lead to aberrant Th2 development in these disease conditions. This study was intended to examine how IL-12-producing ability might associate with allergic diseases as a function of age. IL-12 production by monocytes at various ages was assessed in patients with bronchial asthma and/or atopic dermatitis (n = 100) in comparison with non-allergic control subjects (n = 144). Whole blood cells were stimulated with lipopolysaccharide (LPS) after priming with IFN-gamma, then intracellular cytokine expression of IL-12 and IL-8 as a control cytokine of CD14-positive cells was assessed by flow cytometric analysis. In the control subjects, the ability of monocytes to produce IL-12 was negligible at birth and gradually increased with advancing age, whereas IL-8 production was intense throughout the human life. At more than 7 years of age, IL-12 production of patients with allergic diseases was significantly lower compared with that of control subjects. The unexpected finding was that infants and children below 6 years of age with allergic diseases tended to produce more IL-12 compared with age-matched controls. In this young group, it was noted that enhanced IL-12 production by monocytes was especially observed in allergic patients with specific IgE antibodies against some food allergens. Significant inverse relationships between serum IgE levels and IL-12-producing ability were found in the teenage and adult groups, but not in the younger children. IL-12 appeared to play different roles in the pathogenesis of allergic diseases between younger and older ages.
MeSH Terms
- Adolescent
- Adult
- Aging
- Asthma
- Child
- Child, Preschool
- Dermatitis, Atopic
- Female
- Flow Cytometry
- Humans
- Hypersensitivity
- Infant
- Infant, Newborn
- Interferon-gamma
- Interleukin-12
- Interleukin-8
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Male
- T-Lymphocytes
- Th1 Cells
- Th2 Cells
CD14, a myeloid cell marker and LPS receptor has been acclaimed to play a role in development and manifestation of atopic allergy, as the gene encoding CD14 is located in a chromosomal region linked to total IgE levels and atopic disease. To investigate the levels of soluble (s) and membrane bound (m) CD14 in cord blood and at 2 years of age from children with atopic or non-atopic mothers and relate these parameters to atopy development at 2 years of age. Blood samples were collected at delivery (cord blood) and at 2 years of age among infants with atopic (n = 41) and non-atopic (n = 32) mothers. Blood samples were also obtained from mothers at the same occasions. Levels of sCD14 and total IgE were measured in plasma, and percentages of CD14 cells were measured in cord and peripheral blood mononuclear cells. We observed significant differences in sCD14 levels in cord blood, where children with atopic mothers had the highest levels. The same pattern could be observed in the mothers at delivery. At 2 years of age no significant differences in sCD14 levels were observed between children with atopic mothers and children with non-atopic mothers and no association between sCD14 and atopic disease was found. Further, we observed large differences in sCD14 and mCD14 with respect to age, where newborns displayed a higher frequency of CD14 cells compared with the 2-year-olds and the mothers. The reverse was observed for sCD14, with significantly lower values in cord blood than those seen in the 2-year-olds and mothers. Based on our findings, we suggest that CD14 could be involved in the regulation of IgE production, but that it might also be important for the maturation and development of the neonatal immune system.
MeSH Terms
- Adult
- Aging
- Biomarkers
- Case-Control Studies
- Child, Preschool
- Enzyme-Linked Immunosorbent Assay
- Erythrocyte Membrane
- Female
- Fetal Blood
- Humans
- Hypersensitivity, Immediate
- Immunoglobulin E
- Infant, Newborn
- Lipopolysaccharide Receptors
- Male
- Mothers
- Prospective Studies
- Risk
- Statistics, Nonparametric
Although older subjects are susceptible to thrombosis under septic conditions, the underlying molecular mechanisms have not been fully elucidated. Since elevated plasminogen activator inhibitor-1 (PAI-1) primarily contributes to endotoxin-induced thrombosis, we first compared the induction of PAI-1 by lipopolysaccharide (LPS) between young and aged mice. The higher induction of PAI-1 antigen and mRNA with increased renal glomerular fibrin deposition was observed in LPS-treated aged mice compared to young mice. In situ hybridization analysis showed that the aging-associated induction of PAI-1 mRNA by LPS was pronounced in hepatocytes and in renal glomerular cells. The increased magnitude of the response of aged mice to lower doses of LPS was observed in terms of renal glomerular fibrin deposition and PAI-1 mRNA induction in the tissues. Furthermore, older PAI-1 deficient mice treated with LPS developed much less fibrin deposition in kidneys. Importantly, a larger induction of receptor molecules for LPS (eg, CD14 and Toll-like receptor 4) was demonstrated in LPS-treated aged mice as compared with young mice. The enhanced LPS signaling in aged mice was also demonstrated by the marked induction of nuclear factor-kappaB in the tissues after endotoxin treatment. As a consequence, increases in an inflammatory cytokine, tumor necrosis factor-alpha, were pronounced in plasma and tissues of LPS-treated aged mice. These results emphasize the key role played by PAI-1 in aging-associated deterioration in this thrombosis model, and suggest that the hyperresponse of PAI-1 gene to LPS results from the enhanced LPS signaling and the subsequent inflammatory response in aged mice.
MeSH Terms
- Aging
- Animals
- Dose-Response Relationship, Drug
- Drosophila Proteins
- Fibrin
- Gene Expression Regulation
- Hepatocytes
- In Situ Hybridization
- Kidney Glomerulus
- Kinetics
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Male
- Membrane Glycoproteins
- Mice
- Mice, Inbred C57BL
- NF-kappa B
- Plasminogen Activator Inhibitor 1
- RNA, Messenger
- Receptors, Cell Surface
- Signal Transduction
- Thrombosis
- Toll-Like Receptors
- Tumor Necrosis Factor-alpha
Enhanced stimulus-induced release of pro-inflammatory cytokines by leucocytes may contribute to the pathogenesis of ischaemic stroke. We investigated the lipopolysaccharide-induced release of interleukin-1beta (IL-1beta), IL-6, IL-8, and tumour necrosis factor-alpha (TNF-alpha) in whole blood from 20 patients with a history of ischaemic stroke under the age of 50, 20 patients with a history of cervical artery dissection (CAD) and 21 age- and sex-matched healthy control subjects. Release of IL-8 was higher (P = 0.006) and release of TNF-alpha and IL-6 tended to be higher (P < 0.1) in young stroke patients than in control subjects. No increased release existed in CAD patients. Vascular risk factors or history of infection before stroke did not modify IL-8 production. A common T(250) --> A polymorphism in the IL-8 gene promotor was newly identified but did not correlate with the variability of IL-8 release. The C(260) --> T polymorphism in the gene of the monocytic LPS-receptor CD14--a risk factor for myocardial infarction--was not associated with increased cytokine release. We conclude that high inducible release of IL-8--and possibly of TNF-alpha and IL-6--may contribute to the odds of ischaemic stroke in young adults.
MeSH Terms
- Adult
- Aging
- Aneurysm, Dissecting
- Female
- Humans
- Interleukin-1
- Interleukin-6
- Interleukin-8
- Leukocytes
- Lipopolysaccharides
- Male
- Polymorphism, Genetic
- Risk Factors
- Stroke
- Tumor Necrosis Factor-alpha
Innate immunity not only mediates early host defenses to infection, but also contributes to septic hemodynamic compromise through nitric oxide synthase (NOS2) induction and inhibition of cardiovascular adrenergic responses. Because of increased age-related susceptibility to sepsis, we hypothesized that hearts from old (28-29 months) adult rats would exhibit greater beta-adrenergic hyporesponsiveness than young (6-8 months) following lipopolysaccharide (LPS, 6 mg/kg) with and without interferon gamma (INF-gamma, 5000 units). LPS/INF-gamma depressed baseline dP/dt and isoproterenol-stimulated inotropy in both old and young hearts. beta-adrenergic inotropic ( dP/dt) and lusitropic responses were more depressed in old v young LPS/INF-gamma hearts. Additionally isoproterenol-stimulated cAMP elaboration was less in old (1950 /-160 fmol/min/g) v young (2440 /-170 fmol/min/g, P=0.05) LPS/INF-gamma hearts. LPS alone also depressed basal dP/dt and prolonged myocardial relaxation in old and young hearts, but suppressed isoproterenol dP/dt responses only in old hearts. Depressed beta-adrenergic inotropic responses were augmented with the selective NOS2 inhibitor N-iminoethyl-L-lysine. To establish biochemical mechanisms for this, we tested whether induction of NOS2 and innate immune system receptors (CD14 and Toll-like receptor 4, TLR4) were enhanced in old v young hearts. Induction of myocardial NOS2 and CD14 (not present in control) by LPS/INF-gamma was approximately 2-3-fold greater in old compared to young animals. TLR4 was constitutively expressed in old and young hearts and was unaffected by LPS/INF-gamma. These findings indicate that advanced age is associated with augmented cardiac beta-adrenergic depression and enhanced CD14-NOS2 signaling in response to cytokines. Upregulation of cardiovascular innate immunity may have clinical implications for increased mortality in older individuals with systemic inflammatory response syndromes.
MeSH Terms
- Adrenergic beta-Agonists
- Age Factors
- Aging
- Animals
- Arginine
- Blotting, Western
- Citrulline
- Cyclic AMP
- Cytokines
- Dose-Response Relationship, Drug
- Enzyme Inhibitors
- Immunity
- Interferon-gamma
- Isoproterenol
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Lysine
- Nitric Oxide
- Rats
- Rats, Wistar
- Sepsis
- Signal Transduction
- Up-Regulation
Calorie restriction (CR) is known to prolong the life span and maintain an active immune function in aged mice, but it is still not known if rodents under CR can respond optimally to bacterial infection. We report here on the influence of CR on the response of peritoneal macrophages to lipopolysaccharide, splenic NF-kappaB and NF-interleukin-6 (IL-6) activities, and mortality in polymicrobial sepsis induced by cecal ligation and puncture (CLP). Macrophages from 6-month-old C57BL/6 mice on a calorie-restricted diet were less responsive to lipopolysaccharide, as evidenced by lower levels of IL-12 and IL-6 protein and mRNA expression. Furthermore, in vitro lipopolysaccharide-stimulated macrophages from mice under CR also expressed decreased lipopolysaccharide receptor CD14 levels as well as Toll-like receptor 2 (TLR2) and TLR4 mRNA levels. In addition, the phagocytic capacity and class II (I-A(b)) expression of macrophages were also found to be significantly lower in mice under CR. Mice under CR died earlier (P < 0.005) after sepsis induced by CLP, which appeared to be a result of increased levels in serum of the proinflammatory cytokines tumor necrosis factor alpha and IL-6 and splenic NF-kappaB and NF-IL-6 activation 4 h after CLP. However, mice under CR survived significantly (P < 0.005) longer than mice fed ad libitum when injected with paraquat, a free radical-inducing agent. These data suggest that young mice under CR may be protected against oxidative stress but may have delayed maturation of macrophage function and increased susceptibility to bacterial infection.
MeSH Terms
- Aging
- Animals
- Body Weight
- Cecum
- Drosophila Proteins
- Energy Intake
- Female
- Histocompatibility Antigens Class II
- Interleukin-6
- Ligation
- Lipopolysaccharide Receptors
- Lipopolysaccharides
- Macrophages, Peritoneal
- Membrane Glycoproteins
- Mice
- Mice, Inbred C57BL
- Oxidative Stress
- Peritonitis
- Phagocytosis
- Punctures
- RNA, Messenger
- Receptors, Cell Surface
- Survival Rate
- Toll-Like Receptor 2
- Toll-Like Receptor 4
- Toll-Like Receptors
- Tumor Necrosis Factor-alpha
Previously, a dose-dependent influence of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on CD34 mobilization was demonstrated. In this single-center prospective analysis, 52 healthy donors were investigated to determine the efficacy of intermediate-dose rhG-CSF 2x8 microg/kg donor body weight (bw) and intermediate large volume apheresis (LVA, median 12 l) to mobilize peripheral blood progenitor cells (PBPC) for allogeneic transplantation. The median number of CD34 cells in apheresis products was 0.45% and 2.2x10(6)/kg recipient bw per single apheresis. A total of 5.4x10(6)/kg CD34 cells were collected with two (range: one to three) LVA. In the analysis of donor subgroups, higher peripheral blood (PB) and apheresis results were obtained in male vs female donors; however, donor weight significantly differed in both groups. Heavier donors displayed higher PB and apheresis CD34 counts; however, when CD34 cells/kg were adjusted to a constant bw, similar harvest results were calculated in males and females, demonstrating that gender per se does not, whereas bw does affect apheresis results. Younger donors had significantly higher PB CD34 counts, higher CD34 numbers per single apheresis, increased CFU, more T, B, and CD61 , comparable NK, and less CD14 cells. A correlation analysis of donor age and apheresis results displayed an age-related decline of 0.46x10(6)/kg CD34 cells per decade of donor aging. Cell subsets in apheresis products were CD14 (49%), CD3 (22%), CD4 (13%), CD8 (7%), CD61 (20%), CD19 (5%), and CD16/56 (3%) cells, with increasing CD14 cells and decreasing CD3, CD4, CD8, CD61, CD19, and CD16/56 cells on subsequent days of apheresis. Compared to our previous analysis using high- (2x12 microg) and low-dose (1x10 microg) rhG-CSF for allogeneic PBPC mobilization, the intermediate-dose showed a similar CD34 mobilization potential to 1x10 microg rhG-CSF; however, with use of LVA, two instead of three (p<0.05) aphereses were sufficient to mobilize > or =4x10(6)/kg bw CD34 cells in most donors. Taken together, our results demonstrate that intermediate-dose rhG-CSF sufficiently mobilizes > or =4x10(6)/kg x bw CD34 cells with use of LVA and that especially younger donors display increased CD34 cell numbers.
MeSH Terms
- Adult
- Aging
- Antigens, CD34
- Blood Component Removal
- Blood Donors
- Body Weight
- Cell Count
- Female
- Filgrastim
- Granulocyte Colony-Stimulating Factor
- Hematopoietic Stem Cell Transplantation
- Hematopoietic Stem Cells
- Humans
- Leukocyte Count
- Male
- Middle Aged
- Prospective Studies
- Recombinant Proteins
- Sex Characteristics
- Transplantation, Homologous
Dendritic cells (DC) are powerful antigen presenting cells, which have the unique capacity to stimulate naive T cells. In spite of the well-known decline of T cell function in old age, little information is available on whether DC are also affected by the aging process. This is mainly due to problems with the isolation and purification of DC. Rapid progress in the characterization of DC has been made in recent years, as simple methods to generate large numbers of DC from precursors have been developed. It was the aim of the present study to compare monocyte derived DC from old and young healthy persons. The generation of DC from blood monocytes in response to GM-CSF and IL-4 treatment was similar in cells from young and old persons. The DC population thus obtained had a typical dendritic morphology and expressed DC surface markers, such as HLA class II, CD1a, CD11c, CD54, CD80 and CD86, but not CD14 for a period of up to three weeks in culture. DC from young and old persons produced IL-12 and TNF-alpha and responded equally well to maturation-inducing stimuli. DC maturation was stimulated by purified protein derivative (PPD) of Mycobacterium tuberculosis, whole inactivated influenza virus and by influenza split vaccine, but not by purified viral RNA. When tested for their antigen-presenting capacity, DC from young and old persons were capable of stimulating the proliferation and the cytokine production of T cells. It was of particular interest that CD45RA( ) as well as CD45RO( ) T cells from aged donors were unable to respond to stimulation with influenza proteins presented by monocytes, but were triggered to proliferate and to produce cytokines when antigen was presented by DC. The results demonstrate that DC from old persons (a) may still function as powerful antigen-presenting cells provided the right differentiation and maturation stimuli are present; (b) are capable of mobilizing residual capacity in senescent T cells and (c) may therefore represent a potent tool for immunotherapy and vaccines in old age.
MeSH Terms
- Adult
- Aged
- Aging
- Antigens, Differentiation
- Cell Communication
- Cell Differentiation
- Cytokines
- Dendritic Cells
- Granulocyte-Macrophage Colony-Stimulating Factor
- Humans
- In Vitro Techniques
- Interleukin-4
- Lymphocyte Activation
- Monocytes
- Phenotype
- T-Lymphocytes
We have recently observed that the regression of corpora lutea (CL) in women during the reproductive period of life is accompanied by a diminution of Thy-1 differentiation protein release from vascular pericytes and an accumulation of T lymphocytes and activated macrophages among both degenerating granulosa lutein cells (GLC) and theca lutein cells. These data suggest that the immune system and other stromal factors, representing components of the "tissue control system," may play a role in regression of the CL. We investigated degenerating CL from climacteric women to address the possibility that the decline of immune functions with advancing age may result in incomplete regression of luteal tissue. This could contribute to the altered hormonal profiles and abnormal uterine bleeding that frequently occur during the climacteric. Immunoperoxidase staining and image analysis were used to localize Thy-1 differentiation protein of vascular pericytes, cytokeratin staining of GLC, neural cell adhesion molecule expression by theca lutein cells, CD15 of neutrophils, CD4, CD14, CD68, and leukocyte common antigens of macrophages, and CD3 and CD8 determinants of T lymphocytes. We also investigated the expression of luteinizing hormone receptor (LH receptor) and mitogen activated protein kinases (MAP kinases) in luteal cells. Samples of regressing luteal tissue were obtained during the follicular phase from perimenopausal women (age 45-50) who exhibited prolonged or irregular cycles. For comparison, luteal tissues from women with regular cycles (age 29-45) and CL of pregnancy were also investigated. Corpora lutea of the climacteric women exhibited irregular regression of luteal tissue characterized by a lack of cytoplasmic vacuolization and nuclear pyknosis in GLC, and by a persistence of theca lutein cells exhibiting hyperplasia and adjacent theca externa layers. This was accompanied by a continuing release of Thy-1 differentiation protein from vascular pericytes. Persisting GLC lacked surface expression of macrophage markers (CD4, CD14, CD68 and leukocyte common antigen) as well as nuclear granules exhibiting CD15 of neutrophils, detected in regularly regressing GLC. In addition, such persisting GLC showed weak or no LH receptor expression, and retained the expression of cytokeratin. They also exhibited enhanced staining for MAP kinases. Strong cytoplasmic MAP kinase expression with occasional nuclear translocation was also detected in persisting theca lutein cells, indicating high metabolic activity of these cells. T lymphocytes, although occasionally present in luteal stroma within luteal convolutions, did not invade among persisting GLC and were virtually absent from layers of theca externa and theca lutein cells. These data indicate that the regressing CL in climacteric women may exhibit persistence of luteal cells, perhaps because of age-induced alterations of the immune system and other local stromal homeostatic mechanisms involved in the elimination of luteal cells. Persisting GLC and/or theca lutein cells may exhibit abnormal hormonal secretion that contributes to the alteration of target tissues, such as the endometrium, resulting in abnormal uterine bleeding, hyperplasia, and neoplasia.
MeSH Terms
- Adult
- Aging
- Antigens, CD
- Antigens, Differentiation, Myelomonocytic
- Climacteric
- Endometrium
- Female
- Humans
- Immunohistochemistry
- Keratins
- Lewis X Antigen
- Luteolysis
- Middle Aged
- Neural Cell Adhesion Molecules
- Organ Specificity
- Ovary
- Thy-1 Antigens
As part of a comprehensive study of current and former US Air Force personnel, an extensive assessment of the immune system of 497 normal male subjects was conducted in 1987. Cell surface marker studies for CD2 (total T cells), CD4 (helper T cells), CD8 (suppressor T cells), CD25 (activated T cells), CD20 (total B cells), CD14 (monocytes), and HLA-DR positive cell populations were measured. The CD4/CD8 ratio was also calculated. Functional stimulation assays were also performed using phytohemagglutinin (PHA) and a culture of mixed lymphocytes. Assays of natural killer cells with and without interleukin-2 stimulation were done. In addition to the distribution and range of values for each assay, statistical analyses were performed to determine the effect of age, race, percentage body fat, tobacco use and alcohol consumption on each variable. Age and alcohol consumption had significant correlation with suppressed counts and functions on nearly all variables while tobacco use was associated with stimulation of both T-cell numbers and function. These findings highlight the importance of using age-specific ranges of normal values for these tests of immunity and the need to consider life-style factors in the interpretation of the laboratory assessment of immune status.
MeSH Terms
- Aerospace Medicine
- Aging
- Alcohol Drinking
- Cells, Cultured
- Humans
- Immune System
- Immunoglobulins
- Leukocyte Count
- Life Style
- Lymphocyte Activation
- Lymphocyte Culture Test, Mixed
- Lymphocyte Subsets
- Male
- Military Personnel
- Smoking
- T-Lymphocytes
- United States
Bone marrow cells from 109 patients (median age 60) with newly diagnosed acute myeloid leukaemia (AML) were prospectively immunophenotyped (IP) and the prognostic value of monoclonal antibody (MAB) reactivities was analysed to detect differences in complete remission rates and survival, not only between groups of MAB and - bone marrow cells, but also in cases with or without prominent MAB reactivity as compared to normal BM reactivity of the respective MABs. This approach was based on the assumption that the qualitative expression of antigens is not an all or none phenomenon, but that different degrees of expression of antigens exist. Patients with significantly elevated CD13 (MY7 ) cells in bone marrows (CD13 greater than reference value one standard deviation) (S.D.) showed decreased probability of entering CR (p less than 0.05) and a significantly shorter survival (p less than 0.05). Superior CR rates (p less than 0.05) without difference in long-term survival were seen in patients with low CD33 (MY9) or low HLA-DR expression, while high CD14 (MY4) expression showed a trend towards an adverse factor (p = 0.12). No other antibody reactivities showed differences in CR rates (CD3, CD20, CDw65 (VIM-2) and NAT-9). The more prominent bone marrow expression of CD33 antigen than CD13 (CD33/CD13 greater than 1) correlated to a better chance of entering CR (p = 0.01) and to improved survival (p = 0.002), while the expression of high numbers of VIM-2 cells was a favourable prognostic factor regarding length of survival (p = 0.002). The importance of a high CD33/CD13 ratio as a positive prognostic factor was evaluated using stratified analysis according to age or leucocyte counts at presentation. In both cases, CD33/CD13 was associated with longer survival (age: p = 0.05, leucocyte counts: p = 0.03). A Cox multiparameter analysis revealed that the CD33/CD13 ratio was a favourable prognostic factor (p = 0.03) together with age (p = 0.001) and leucocyte counts in peripheral blood (p less than 0.01). We conclude that establishing the immunologic phenotype can be of prognostic value in cases of AML, especially with regard to the relationship between the CD33 and CD13 antigens.
MeSH Terms
- Acute Disease
- Adolescent
- Adult
- Aged
- Aged, 80 and over
- Aging
- Analysis of Variance
- Antibodies, Monoclonal
- Antigens, Neoplasm
- Antineoplastic Combined Chemotherapy Protocols
- Female
- Humans
- Immunophenotyping
- Leukemia, Myeloid
- Leukocyte Count
- Male
- Middle Aged
- Multivariate Analysis
- Prognosis
- Prospective Studies