CD19
B-lymphocyte antigen CD19 precursor (B-lymphocyte surface antigen B4) (Differentiation antigen CD19) (T-cell surface antigen Leu-12) (CD19 antigen)
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Socioeconomic status (SES), living in poverty, and other social determinants of health contribute to health disparities in the United States. African American (AA) men living below poverty in Baltimore City have a higher incidence of mortality when compared to either white males or AA females living below poverty. Previous studies in our laboratory and elsewhere suggest that environmental conditions are associated with differential gene expression (DGE) patterns in peripheral blood mononuclear cells (PBMCs). DGE have also been associated with hypertension and cardiovascular disease (CVD) and correlate with race and sex. However, no studies have investigated how poverty status associates with DGE between male and female AAs and whites living in Baltimore City. We examined DGE in 52 AA and white participants of the Healthy Aging in Neighborhoods of Diversity across the Life Span (HANDLS) cohort, who were living above or below 125% of the 2004 federal poverty line at time of sample collection. We performed a microarray to assess DGE patterns in PBMCs from these participants. AA males and females living in poverty had the most genes differentially-expressed compared with above poverty controls. Gene ontology (GO) analysis identified unique and overlapping pathways related to the endosome, single-stranded RNA binding, long-chain fatty-acyl-CoA biosynthesis, toll-like receptor signaling, and others within AA males and females living in poverty and compared with their above poverty controls. We performed RT-qPCR to validate top differentially-expressed genes in AA males. We found that KLF6, DUSP2, RBM34, and CD19 are expressed at significantly lower levels in AA males in poverty and KCTD12 is higher compared to above poverty controls. This study serves as an additional link to better understand the gene expression response in peripheral blood mononuclear cells in those living in poverty.
MeSH Terms
- Adult
- Demography
- Female
- Gene Expression Profiling
- Humans
- Longevity
- Male
- Metabolic Networks and Pathways
- Middle Aged
- Monocytes
- Poverty
- Transcriptome
- Urban Population
The SARS-CoV-2 infection has widely spread to become the greatest public health challenge to date, the COVID-19 pandemic. Different fatality rates among countries are probably due to non-standardized records being carried out by local health authorities. The Spanish case-fatality rate is 11.22%, far higher than those reported in Asia or by other European countries. A multicentre retrospective study of demographic, clinical, laboratory and immunological features of 584 Spanish COVID-19 hospitalized patients and their outcomes was performed. The use of renin-angiotensin system blockers was also analysed as a risk factor. In this study, 27.4% of cases presented a mild course, 42.1% a moderate one and for 30.5% of cases, the course was severe. Ages ranged from 18 to 98 (average 63). Almost 60 % (59.8%) of patients were male. Interleukin 6 was higher as severity increased. On the other hand, CD8 lymphocyte count was significantly lower as severity grew and subpopulations CD4, CD8, CD19, and NK showed concordant lowering trends. Severity-related natural killer percent descents were evidenced just within aged cases. A significant severity-related decrease of CD4 lymphocytes was found in males. The use of angiotensin-converting enzyme inhibitors was associated with a better prognosis. The angiotensin II receptor blocker use was associated with a more severe course. Age and age-related comorbidities, such as dyslipidaemia, hypertension or diabetes, determined more frequent severe forms of the disease in this study than in previous literature cohorts. Our cases are older than those so far reported and the clinical course of the disease is found to be impaired by age. Immunosenescence might be therefore a suitable explanation for the hampering of immune system effectors. The adaptive immunity would become exhausted and a strong but ineffective and almost deleterious innate response would account for COVID-19 severity. Angiotensin-converting enzyme inhibitors used by hypertensive patients have a protective effect in regards to COVID-19 severity in our series. Conversely, patients on angiotensin II receptor blockers showed a severer disease.
Keywords
- ACE2
- C-reactive protein
- COVID-19
- Immunity
- Immunosenescence
- Interleukin-6
- Lymphocytes
- Renin-angiotensin system
- Severe acute respiratory syndrome coronavirus 2
- Spain
Ageing is often characterised by nutritional deficiencies and functional alterations of the digestive and immune system. The aim of the present study was to analyse the impact of consumption of conventional milk with A1/A2 beta-casein, compared to milk containing only the A2 beta-casein variant, characterised by a protein profile favouring gut health. Twenty-four ageing Balb-c mice (20 months old) were fed for 4 weeks, with either a control diet (CTRL), a diet supplemented with bovine milk containing A1/A2 beta-casein (A1A2) or a diet containing A2/A2 beta-casein (A2A2). Lymphocyte subpopulations, enzymatic activities, cytokine secretion, gut morphology and histopathological alterations were measured in different gut segments, while short-chain fatty acids (SCFAs) content and microbiota composition were evaluated in faecal samples. The A2A2 group showed higher content of faecal SCFAs (in particular, isobutyrate) of intestinal CD4 and CD19 lymphocytes in the intraepithelial compartment and improved villi tropism. The A1A2 group showed higher percentages of intestinal TCRγδ lymphocytes. Faecal microbiota identified [i]Deferribacteriaceae[/i] and [i]Desulfovibrionaceae[/i] as the most discriminant families for the A2A2 group, while [i]Ruminococcaceae[/i] were associated to the A1A2 group. Taken together, these results suggest a positive role of milk, in particular when containing exclusively A2 beta-casein, on gut immunology and morphology of an ageing mice model.
Keywords
- A2 beta-casein
- SCFAs
- elderly
- gut inflammation
- gut microbiota
- gut morphology
- immunosenescence
Treatment with chimeric antigen receptor (CAR)-modified T cells targeting CD19 has proved successful in patients with relapsed/refractory B cell malignancies. However, long-term follow-up indicates that remission in a substantial proportion of patients is not sustainable. Most patients that experience recurrence have tumors and lost the CAR-T cells. To maintain the activity of CAR-T cells, Raji-B-NDG mice were treated sequentially with CAR-T-19 cells and homologous cells expressing human CD19 to promote expansion of CAR-T cells. Sequential treatment of mice with CAR-T-19 cells followed by Raji tumor cells led to marked prolongation of survival. The best case scenario after sequential treatment was a survival time of more than 200 days; the average survival time of mice in the non-sequential treatment group was 80 days. We treated mice with autologous CD19-modified T cells after initial treatment with CAR-T-19 cells. The overall survival and recurrence-free survival times of mice receiving sequential treatment were significantly longer. The percentages of CAR T cells in peripheral blood increased. Sequential therapy with autologous CAR-T-19 and aT19 cells provides a new strategy for generating memory CAR-T cells, which may lead ultimately to increased clinical efficacy.
MeSH Terms
- Animals
- Antigens, CD19
- Cell Line, Tumor
- Combined Modality Therapy
- Disease-Free Survival
- HEK293 Cells
- Healthy Volunteers
- Humans
- Immunologic Memory
- Immunotherapy, Adoptive
- Longevity
- Lymphoma, B-Cell
- Mice
- Neoplasm Recurrence, Local
- Receptors, Chimeric Antigen
- Recombinant Proteins
- Remission Induction
- T-Lymphocytes
- Time Factors
- Transduction, Genetic
- Transplantation, Autologous
- Xenograft Model Antitumor Assays
Keywords
- Autologous CD19 T cells
- Chimeric antigen receptor
- Memory T cells
- Sequential therapy
Cellular senescence has been thought to be an important barrier to tumor formation. Recent studies have shown that stress-induced premature senescence (SIPS) can promote partial tumor invasion, but how SIPS affects diffuse large B-cell lymphoma (DLBCL) remains inconclusive. This study aimed to address that issue. The immunophenotype of the LY8 cell line was measured with flow cytometry. SIPS induced by tert-butyl hydroperoxide (tBHP) was detected by senescence β-galactosidase staining. Cell proliferation was analyzed with CCK8 and expression levels of ARHGAP18 ([i]SENEX[/i] gene-encoding protein), p16/p21, and Rb/pRb were measured with western blot. LY8 cells were transfected with [i]SENEX[/i]-SiRNA/NC and verified by western blot. Our results suggested that the immunophenotype of the LY8 cell line is CD19-, CD20-, and CD10-positive and the immunoglobulin light chain is the kappa type. The cellular senescence model of DLBCL could be successfully induced by 30 μM tBHP. ARHGAP18, p21, p16, and Rb protein levels were significantly increased but the level of pRb expression was decreased in the SIPS group compared with other groups. Meanwhile, the proliferation rate was increased in the SIPS group more than other tBHP groups. Furthermore, the expressions of p21 and p16 were significantly decreased in the [i]SENEX[/i]-SiRNA group compared with the negative control group. SIPS formation activates ARHGAP18 and the p16/Rb pathway and promotes DLBCL cell proliferation. Furthermore, [i]SENEX[/i] activates the p16 pathway in DLBCL. SIPS promotes proliferation by activating [i]SENEX[/i] and the p16/Rb pathway in DLBCL. [i]SENEX[/i]-related SIPS may serve as an important target for relapsed/refractory DLBCL therapy.
MeSH Terms
- Cell Culture Techniques
- Cell Line, Tumor
- Cell Proliferation
- Cellular Senescence
- Cyclin-Dependent Kinase Inhibitor p16
- Humans
- Lymphoma, Large B-Cell, Diffuse
- Retinoblastoma
- Transfection
Keywords
- Stress-induced premature senescence
- Proliferation
- SENEX
- p16
- Rb/pRb
- Diffuse large B-cell lymphoma
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
Hypovitaminosis D is associated with age-related illnesses, including hypertension, cardiovascular disease (CRVD), cerebrovascular disease (CAD) and type 2 diabetes mellitus (T2DM). In our retrospective observational study, blood samples of elderly healthy controls (n = 461) and patients with age-related diseases (n = 8,621) were subjected to flow-cytometry in order to determine correlations between age-related diseases and cluster of differentiation 4 (CD4), CD8, CD3, and CD19 lymphocyte markers, as well as serum levels of 25-hydroxyvitamin D (25(OH)D ) and 25-hydroxyvitamin D (25(OH)D ). More than 70% of the patients in each disease group had total vitamin D < 20 ng/mL (P < 0.001). In CRVD patients, CD3 and CD19 correlated (P < 0.05) with 25(OH)D . In CAD patients, CD8, CD4, CD19 and CD4/CD8 correlated (P < 0.05) with 25(OH)D , and CD8 correlated (P < 0.05) with 25(OH)D . In T2DM and hypertension patients, CD8, CD3, CD19 and CD4/CD8 correlated with 25(OH)D . Progressive trends (P < 0.05) towards increased CD8 and CD4/CD8 were observed in vitamin-D-deficient T2DM and hypertension patients. Significant differences (P < 0.05) in CD8 were observed in vitamin-D-deficient CAD patients, whereas significant differences (P < 0.05) in CD8 and CD19 were observed in CRVD patients. Higher CD8 and CD4/CD8 in 25(OH)D-deficient T2DM and hypertension patients suggested a Th1 lymphocyte profile induction. Increases in CD8-positive lymphocytes suggested a similar, less pronounced effect in vitamin-D-deficient CRVD and CAD patients.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Biomarkers
- Cardiovascular Diseases
- Case-Control Studies
- Cerebrovascular Disorders
- Diabetes Mellitus, Type 2
- Female
- Humans
- Hypertension
- Lymphocyte Subsets
- Male
- Retrospective Studies
- Vitamin D
- Vitamin D Deficiency
- Vitamins
Normal B lymphoid maturation occurs in bone marrow (BM) throughout life, but immature B-cell progenitors (BCPs) are more numerous in children than in adults. To assess the normal values according to age became important as BCPs are decreased in myelodysplastic syndromes and have been considered an important diagnostic and prognostic feature in these clonal disorders. in a multicenter retrospective study from the Brazilian Group of Flow Cytometry we analyzed the variation of BCPs in normal BM according to age and technical peculiarities of each laboratory. We analysed of 45 BM donors and 89 cases examined for elucidation of transitory reactive cytopenias presenting a normal BM immunophenotyping. BCPs were enumerated as CD19 /CD34 /CD45 /CD10 cells (panel 1) or CD19 /CD34 /CD45 cells (panel 2) among the total nucleated non-erythroid cells and as percentage of CD34 cells. we included 134 cases. Panel 1 was applied in 88 cases and panel 2 was used in 46. Age range: 10 months to 89 years. In a multiple regression, % BCPs/total nucleated cells was an exponential function of age. Age explained alone 49.4% of the variance, while 'panel used' explained 1.8% and 'laboratory' explained 0.7%. Age explained only 24.9% of the variance of BCPs/CD34 cells. in normal individuals, BM B-cell precursors varied mainly according to age, but were also dependent on technical peculiarities of operators and equipments. Analysis by phenotype and as percentage of total nucleated cells was more accurate and less susceptible to variation than evaluating % BCPs/total CD34 cells. © 2017 International Clinical Cytometry Society.
MeSH Terms
- Adolescent
- Adult
- Age Factors
- Aged
- Aged, 80 and over
- Aging
- Brazil
- Child
- Child, Preschool
- Flow Cytometry
- Humans
- Infant
- Middle Aged
- Myelodysplastic Syndromes
- Precursor Cells, B-Lymphoid
- Reference Values
- Retrospective Studies
- Young Adult
Keywords
- age variation
- bone marrow
- cytometry
- hematogones
Aging has a strong impact on the activity of the immune system, enhancing susceptibility to pathogens and provoking a predominant pre-inflammatory status, whereas dampening responses to vaccines in humans and mice. Here, we demonstrate a loss of marginal zone B lymphocytes (MZ, CD19 CD45R CD21 CD23 ) and a decrease of naive B cells (CD19 IgD ), whereas there is an enhancement of a CD19 CD45R innate-like B cell population (B1REL) and the so-called aged B cell compartment (ABC, CD45R CD21 CD23 CD5 CD11b ) in aged senescence-accelerated (SAMP8) mice but not in aged senescence-resistant (SAMR1) mice. These changes in aged SAMP8 mice were associated with lower IgG isotype levels, displaying low variable gene usage repertoires of the immunoglobulin heavy chain (V ) diversity, with a diminution on IgG1-memory B cells (CD11b Gr1 CD138 IgM IgD CD19 CD38 IgG1 ), an increase in T follicular helper (T , CD4 CXCR5 PD1 ) cell numbers, and an altered MOMA-1 (metallophilic macrophages) band in primary follicles. LPS-mediated IgG1 responses were impaired in the B1REL and ABC cell compartments, both in vitro and in vivo. These data demonstrate the prominent changes to different B cell populations and in structural follicle organization that occur upon aging in SAMP8 mice. These novel results raise new questions regarding the importance of the cellular distribution in the B cell layers, and their effector functions needed to mount a coordinated and effective humoral response.
MeSH Terms
- Aging
- Animals
- Antigens, CD
- B-Lymphocytes
- Cell Death
- Cell Proliferation
- Gene Expression Regulation, Developmental
- IgG Deficiency
- Immunity, Humoral
- Immunity, Innate
- Immunoglobulin D
- Immunoglobulin G
- Immunoglobulin Heavy Chains
- Immunoglobulin M
- Immunologic Memory
- Lipopolysaccharides
- Mice, Inbred C57BL
- Mice, Transgenic
- Primary Cell Culture
- Signal Transduction
- Spleen
- T-Lymphocytes, Helper-Inducer
Many reports have shown that various kinds of stem cells have the ability to recover premature ovarian aging (POA) function. Transplantation of human amniotic epithelial cells (hAECs) improves ovarian function damaged by chemotherapy in a mice model. Understanding of how to evaluate the distinct effects of adult stem cells in curing POA and how to choose stem cells in clinical application is lacking. To build a different degrees of POA model, mice were administered different doses of cyclophosphamide: light dose (70 mg/kg, 2 weeks), medium dose (70 mg/kg, 1 week; 120 mg/kg, 1 week), and high dose (120 mg/kg, 2 weeks). Enzyme-linked immunosorbent assay detected serum levels of sex hormones, and hematoxylin and eosin staining allowed follicle counting and showed the ovarian tissue structure. DiIC (5)-DS was employed to label human amniotic mesenchymal stem cells (hAMSCs) and hAECs for detecting the cellular retention time in ovaries by a live imaging system. Proliferation of human ovarian granule cells (ki67, AMH, FSHR, FOXL2, and CYP19A1) and immunological rejection of human peripheral blood mononuclear cells (CD4, CD11b, CD19, and CD56) were measured by flow cytometry (fluorescence-activated cell sorting (FACS)). Distinction of cellular biological characteristics between hAECs and hAMSCs was evaluated, such as collagen secretory level (collagen I, II, III, IV, and VI), telomerase activity, pluripotent markers tested by western blot, expression level of immune molecules (HLA-ABC and HLA-DR) analyzed by FACS, and cytokines (growth factors, chemotactic factors, apoptosis factors, and inflammatory factors) measured by a protein antibody array methodology. After hAMSCs and hAECs were transplanted into a different degrees of POA model, hAMSCs exerted better therapeutic activity on mouse ovarian function in the high-dose administration group, promoting the proliferation rate of ovarian granular cells from premature ovarian failure patients, but also provoking immune rejection. Meanwhile, our results showed that the biological characteristics of hAMSCs were superior to hAECs, but not to expression of immune molecules. These results suggest that hAMSCs are a more effective cell type to improve ovarian function than hAECs. Meanwhile, this distinct effect is attributable to cellular biological characteristics of hAMSCs (telomerase activity, expression level of pluripotent markers, cytokine and collagen secretion) that are superior to hAECs, except for immunological rejection. Sufficient consideration of cell properties is warranted to move forward to more effective clinical therapy.
MeSH Terms
- Amnion
- Animals
- Disease Models, Animal
- Epithelial Cells
- Female
- Heterografts
- Humans
- Mice
- Mice, Inbred ICR
- Primary Ovarian Insufficiency
Keywords
- Cellular biological characteristics
- Human amniotic epithelial cells
- Human amniotic mesenchymal stem cells
- Premature ovarian aging
Aging is associated with a decline in the normal functioning of the immune system. Several studies described the relationship between immunological alterations, including immunosenescence and inflammation, and aging or age-related outcomes, such as sarcopenia, depression, and neurodegenerative disorders. Physical activity is known to improve muscle function and to exert a number of benefits on older adult health, including reduced risk for heart and metabolic system chronic diseases. However, the positive influence of physical activity on the immune system has not been elucidated. In order to shed light on the role of physical activity in immune responses of older individuals, a number of immunological parameters comprising % lymphocyte subsets (CD3 , CD4 , CD8 , CD19 , and CD16 56 ) and serum levels of neopterin and tryptophan metabolism products were evaluated in peripheral blood samples of older adults performing normal (N = 170) or reduced (N = 89) physical activity. In addition, the potential influence of other clinical and epidemiological factors was also considered. Results showed that subjects with reduced physical activity displayed significantly higher levels of CD4 /CD8 ratio, kynurenine/tryptophan ratio, and serum neopterin, along with lower %CD19 cells and tryptophan concentrations. Further, some immunological biomarkers were associated with cognitive impairment and functional status. These data contribute to reinforce the postulation that physical activity supports healthy aging, particularly by helping to protect the immunological system from aging-related changes.
MeSH Terms
- Activities of Daily Living
- Age Factors
- Aged
- Aged, 80 and over
- Aging
- Biomarkers
- CD4-CD8 Ratio
- Cognitive Dysfunction
- Exercise
- Female
- Humans
- Immune System
- Kynurenine
- Lymphocyte Subsets
- Male
- Neopterin
- Surveys and Questionnaires
- Tryptophan
Mice transgenic for human CD19 have been an important animal model to help understand the role of this molecule in B lymphocyte function. Previously, no lifetime studies had been performed to understand the effects of this CD19 over expression on the survival or spontaneous pathology within the C57BL/6J background strain. We conducted a lifetime study with interim sacrifices to understand the transgenic effects on clinical signs, body weight, survival, and spontaneous pathology. Blood and urine samples were collected from select animals at various time points during the study for measurement of clinical pathology parameters and groups of animals were euthanized and examined at predetermined intervals. There was fair survival with some animals living to 108 weeks of age. Clinical pathology evaluations revealed a declining red cell mass with a regenerative anemia, increasing total white blood cell counts and decreasing glucose level. Total protein, albumin, and globulin levels increased to 52 weeks of age and then declined to or below baseline with advancing age. Increased urinary microalbumin levels correlated with the severity of a glomerulopathy at 76 and 84 weeks of age. Mean body weight increased through 70 weeks and then declined to weights similar to week 28 at 108 weeks. Macroscopic observations included pale kidneys, enlarged seminal vesicles, and enlarged spleens (at 108 weeks of age). The most common neoplasms in this study were bronchiolar alveolar adenomas in the lung, histiocytic sarcoma in several different tissues, and hepatocellular adenomas. The most common non-neoplastic lesions were renal glomerulopathy, and pulmonary lymphocytic infiltrates with increased numbers of alveolar macrophages.
MeSH Terms
- Aging
- Animals
- Antigens, CD19
- Blood Cell Count
- Blood Chemical Analysis
- Blood Glucose
- Body Weight
- Kidney Diseases
- Male
- Mice, Inbred C57BL
- Mice, Transgenic
- Neoplasms
- Organ Size
Keywords
- Aged
- CD19
- Inebilizumab
- huCD19 Tg
Age-related arterial inflammation is associated with dysfunction of the arteries and increased risk for cardiovascular disease. To determine if aging increases arterial immune cell infiltration as well as the populations of immune cells principally involved, we tested the hypothesis that large elastic and resistance arteries in old mice would exhibit increased immune cell infiltration compared to young controls. Additionally, we hypothesized that vasoprotective lifestyle interventions such as lifelong caloric restriction or 8weeks of voluntary wheel running would attenuate age-related arterial immune cell infiltration. The aorta and mesenteric vasculature with surrounding perivascular adipose was excised from young normal chow (YNC, 4-6months, n=10), old normal chow (ONC, 28-29months, n=11), old caloric restricted (OCR, 28-29months, n=9), and old voluntary running (OVR, 28-29months, n=5) mice and digested to a single cell suspension. The cells were then labeled with antibodies against CD45 (total leukocytes), CD3 (pan T cells), CD4 (T helper cells), CD8 (cytotoxic T cells), CD19 (B cells), CD11b, and F4/80 (macrophages) and analyzed by flow cytometry. Total leukocytes, T cells (both CD4 and CD8 subsets), B cells, and macrophages in both aorta and mesentery were all 5- to 6-fold greater in ONC compared to YNC. Age-related increases in T cell (both CD4 and CD8 ), B cell, and macrophage infiltration in aorta were abolished in OCR mice. OVR mice exhibited 50% lower aortic T cell and normalized macrophage infiltration. B cell infiltration was not affected by VR. Age-related mesenteric CD8 T cell and macrophage infiltration was normalized in OCR and OVR mice compared to young mice, whereas B cell infiltration was normalized by CR but not VR. Splenic CD4 T cells from ONC mice exhibited a 3-fold increase in gene expression for the T helper (Th) 1 transcription factor, Tbet, and a 4-fold increase in FoxP3, a T regulatory cell transcription factor, compared to YNC. Splenic B cells and mesenteric macrophages from old mice exhibited decreased proinflammatory cytokine gene expression regardless of treatment group. These results demonstrate that aging is associated with infiltration of immune cells around both the large-elastic and resistance arteries and that the vasoprotective lifestyle interventions, CR and VR, can ameliorate age-related arterial immune cell infiltration.
MeSH Terms
- Aging
- Animals
- Arteries
- Caloric Restriction
- Leukocytes
- Macrophages
- Male
- Mice
- Physical Conditioning, Animal
- Vascular Diseases
Keywords
- Aorta
- B cells
- Inflammation
- Macrophages
- Mesentery
- T cells
B cells undergo maturation and class-switching in response to antigen exposure and T-cell help. Early B-cell differentiation has not been defined in patients with early-onset atopic dermatitis (AD). We sought to define the frequency of B-cell subsets associated with progressive B-cell maturation and IgE class-switching. We studied 27 children and 34 adults with moderate-to-severe AD (mean SCORAD score, 55 and 65, respectively) and age-matched control subjects (15 children and 27 adults). IgD/CD27 and CD24/CD38 core gating systems and an 11-color flow cytometric panel were used to determine the frequencies of circulating B-cell subsets. Serum total and allergen-specific IgE (sIgE) levels were measured by using ImmunoCAP. Compared with adults, children showed T-cell predominance in the skin. Circulating CD19 CD20 B-cell counts were lower in patients with pediatric AD than in control subjects (24% vs 33%, P = .04), whereas CD3 T-cell counts were higher (62% vs 52%, P = .05). A decreased B-cell/T-cell lymphocyte ratio with age was observed only in pediatric control subjects (r = -0.48, P = .07). In pediatric patients with AD, a positive correlation was observed between B-cell/T-cell ratio and nonswitched memory B-cell counts (r = 0.42, P = .03). Higher frequencies of positive sIgE levels were seen in pediatric patients with AD (P < .0001). Diverse sIgE levels correlated with SCORAD scores and age of pediatric patients with AD (P < .01). Positive correlations were observed between activated B-cell and memory T-cell counts (P < .02). In patients with AD, IgE sensitization to most allergens clustered with age, T 1, T 2, total IgE levels, and B-cell memory subsets. Peripheral B and T cells are altered in pediatric patients with early AD, but T cells predominate in skin lesions.
MeSH Terms
- Adult
- Aging
- Allergens
- B-Lymphocyte Subsets
- Child, Preschool
- Dermatitis, Atopic
- Female
- Humans
- Immunoglobulin E
- Infant
- Male
- Middle Aged
- Skin
- T-Lymphocytes
Keywords
- Atopic dermatitis
- B cells
- T cells
- allergen-specific IgE
- atopic march
The ataxia telangiectasia mutated (ATM)-interacting protein ATMIN mediates noncanonical ATM signaling in response to oxidative and replicative stress conditions. Like ATM, ATMIN can function as a tumor suppressor in the hematopoietic system: deletion of Atmin under the control of CD19-Cre results in B-cell lymphomas in aging mice. ATM signaling is essential for lymphopoiesis and hematopoietic stem cell (HSC) function; however, little is known about the role of ATMIN in hematopoiesis. We thus sought to investigate whether the absence of ATMIN would affect primitive hematopoietic cells in an ATM-dependent or -independent manner. Apart from its role in B-cell development, we show that ATMIN has an ATM-independent function in the common myeloid progenitors (CMPs) by deletion of Atmin in the entire hematopoietic system using Vav-Cre. Despite the lack of lymphoma formation, ATMIN-deficient mice developed chronic leukopenia as a result of high levels of apoptosis in B cells and CMPs and induced a compensatory mechanism in which HSCs displayed enhanced cycling. Consequently, ATMIN-deficient HSCs showed impaired regeneration ability with the induction of the DNA oxidative stress response, especially when aged. ATMIN, therefore, has multiple roles in different cell types, and its absence results in perturbed hematopoiesis, especially during stress conditions and aging.
MeSH Terms
- Aging
- Animals
- Apoptosis
- Ataxia Telangiectasia Mutated Proteins
- B-Lymphocytes
- Chronic Disease
- Gene Deletion
- Hematopoiesis
- Hematopoietic Stem Cells
- Leukopenia
- Mice
- Mice, Knockout
- Oxidative Stress
- Transcription Factors
Physiological ageing is accompanied by decline in immune system function and immune alteration during ageing increases susceptibility to infections. We retrospectively analysed the data for complete blood count (CBC) and lymphocyte subsets from infant to elderly age groups to determine changes during ageing. Data from dual-platform flow cytometry and CBC were analysed to determine the percentage (%) and absolute cell counts (Abs) of peripheral blood lymphocyte subsets (CD3, CD4, CD8, CD19 and CD56 16 cells) in infants (1 month to 1 year), children (1 year to 6 years), adolescents (12 years to 18 years), adults (21 years to 50) and elderly (70 years to 92 years). Differences in plasma cytokine levels in adults and elderly were also analysed using Randox system. Comparisons among age groups from infants through adults revealed progressive declines in the percentage of total lymphocytes and absolute numbers of T and B cells. The NK cells declined from infancy to adulthood but increased in elderly participants. The percentages of T cells increased with age from infant to adulthood and then declined. Pro-inflammatory cytokines, TNF-α and IL-6, were higher in elderly people compared to adults. The elderly group had significantly higher levels of monocyte chemoattractant protein-1 (MCP-1) and lower levels of epidermal growth factor (EGF) compared to adults. Our findings confirm and extend earlier reports on age-related changes in lymphocyte subpopulations and data generated from this study is useful for clinicians and researchers, patient management in various age groups for the interpretation of disease-related changes, as well as therapy-dependent alterations.
MeSH Terms
- Adolescent
- Adult
- Age Factors
- Aged
- Aged, 80 and over
- Aging
- B-Lymphocytes
- Blood Cell Count
- Chemokine CCL2
- Child
- Child, Preschool
- Cytokines
- Epidermal Growth Factor
- Female
- Flow Cytometry
- Humans
- Infant
- Infant, Newborn
- Interleukin-6
- Killer Cells, Natural
- Lymphocyte Count
- Lymphocyte Subsets
- Male
- Middle Aged
- Retrospective Studies
- T-Lymphocytes
- Tumor Necrosis Factor-alpha
- Young Adult
Very early onset inflammatory bowel disease (VEO-IBD), IBD diagnosed at 5 years of age or younger, frequently presents with a different and more severe phenotype than older-onset IBD. We investigated whether patients with VEO-IBD carry rare or novel variants in genes associated with immunodeficiencies that might contribute to disease development. Patients with VEO-IBD and parents (when available) were recruited from the Children's Hospital of Philadelphia from March 2013 through July 2014. We analyzed DNA from 125 patients with VEO-IBD (age, 3 wk to 4 y) and 19 parents, 4 of whom also had IBD. Exome capture was performed by Agilent SureSelect V4, and sequencing was performed using the Illumina HiSeq platform. Alignment to human genome GRCh37 was achieved followed by postprocessing and variant calling. After functional annotation, candidate variants were analyzed for change in protein function, minor allele frequency less than 0.1%, and scaled combined annotation-dependent depletion scores of 10 or less. We focused on genes associated with primary immunodeficiencies and related pathways. An additional 210 exome samples from patients with pediatric IBD (n = 45) or adult-onset Crohn's disease (n = 20) and healthy individuals (controls, n = 145) were obtained from the University of Kiel, Germany, and used as control groups. Four hundred genes and regions associated with primary immunodeficiency, covering approximately 6500 coding exons totaling more than 1 Mbp of coding sequence, were selected from the whole-exome data. Our analysis showed novel and rare variants within these genes that could contribute to the development of VEO-IBD, including rare heterozygous missense variants in IL10RA and previously unidentified variants in MSH5 and CD19. In an exome sequence analysis of patients with VEO-IBD and their parents, we identified variants in genes that regulate B- and T-cell functions and could contribute to pathogenesis. Our analysis could lead to the identification of previously unidentified IBD-associated variants.
MeSH Terms
- Adolescent
- Adult
- Aging
- Antigens, CD19
- Cell Cycle Proteins
- Child
- Child, Preschool
- Exome
- Female
- Gene Frequency
- Genetic Association Studies
- Genetic Predisposition to Disease
- Germany
- High-Throughput Nucleotide Sequencing
- Humans
- Immunologic Deficiency Syndromes
- Infant
- Infant, Newborn
- Inflammatory Bowel Diseases
- Interleukin-10 Receptor alpha Subunit
- Male
- Middle Aged
- Mutation
- Sequence Analysis, DNA
Keywords
- CVID
- Common Variable Immune Deficiency
- IBD
- Inherited Defects
- Innate and Adaptive Immunity
To observe the differentiation ability of effector B cells induced by soluble egg antigen (SEA) and soluble adult worm antigen (SWAP) of Schistosoma japonicum. The mouse spleen mononuclear cells and CDl9 B cells sorted by beadg were stimulated by SEA, SWAP and lipopolysaccharide (LPS), respectively. The ratios of CD19 IL-6 cells and CD19 IFN-gamma cells were analyzed by flow cytometry after 72 hours. At the same time, the cytokines IL-6 and IFN-gamma levels in the cultured supernatants were detected by ELISA. The mouse was immunized with the mixture of SEA or SWAP or LPS and the incomplete Freund' s adjuvant for three times, respectively. The mouse spleen mononuclear cells were isolated at the seventh day after the last immunization. The ratios of CD19 IL-6 cells and CD19 IFN-gama cells were analyzed, and the cytokines IL-6 and IFN-gamma levels in the culture supernatants were detected. The ratio of CD19 IL-6 cells in spleen mononuclear cells and splenic B cells was significantly increased in the groups stimulated by SEA and LPS (P < 0.05), and the cytokines IL-6 level in the CD193 cells culture supernatants were significantly increased (P < 0.01). Furthermore, the ratio of CD19 IL-6 cells and the cytokines IL-6 level were significantly increased in the SEA immunized group (P < 0.01). SWAP could induce a significantly higher ratio of the CD19 IFN-gamma cells in spleen cells, instead of in splenic CD19 B cells (P < 0.05). The CD19 IFN-gamma cells and the cytokine IFN-gamma level in the culture supernatants in the SWAP immunized group were significantly higher than those in the SEA and PBS immunized groups (P C 6.01). SEA preferentially induces the increase of CDl9 IL-6 cells in mouse spleen cells; while SWAP preferentially induces the CD19 IFN-gamma cells' production of mouse spleen cells, depending on the effects of other immune cells.
MeSH Terms
- Aging
- Animals
- Antigens, Helminth
- B-Lymphocytes
- Cell Differentiation
- Female
- Interferon-gamma
- Interleukin-6
- Mice
- Mice, Inbred C57BL
- Ovum
- Schistosoma japonicum
Depression is one of the most frequently missed diagnoses in elderly people, with obvious negative effects on quality of life. Various studies have shown that long chain omega-3 polyunsaturated fatty acids (n-3 PUFA) may be useful in its management. Our objective was to evaluate whether a supplement containing n-3 PUFA improves depressive symptoms in depressed elderly patients, and whether the blood fatty acid pattern is correlated with these changes. The severity of depressive symptoms according to the Geriatric Depression Scale (GDS), blood fatty acid composition and erythrocyte phospholipids were analyzed in 46 depressed females aged 66-95y, diagnosed with depression according to DSMIV, within the context of a randomized, double-blind, placebo-controlled trial. 22 depressed females were included in the intervention group (2.5 g/day of n-3 PUFA for 8 weeks), and 24 in the placebo group. We also measured immunological parameters (CD2, CD3, CD4, CD8, CD16, CD19 and cytokines (IL-5, IL-15). The mean GDS score and AA/EPA ratio, in whole blood and RBC membrane phospholipids, were significantly lower after 2 months supplementation with n-3 PUFA. A significant correlation between the amelioration of GDS and the AA/EPA ratio with some immunological parameters, such as CD2, CD19, CD4, CD16 and the ratio CD4/CD8, was also found. Nevertheless, omega-3 supplementation did not significantly improve the studied immunological functions. n-3 PUFA supplementation ameliorates symptoms in elderly depression. The n-3 PUFA status may be monitored by means of the determination of whole blood AA/EPA ratio.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Antidepressive Agents
- Antigens, CD
- Arachidonic Acid
- Cytokines
- Depression
- Diagnostic and Statistical Manual of Mental Disorders
- Dietary Supplements
- Double-Blind Method
- Eicosapentaenoic Acid
- Erythrocyte Membrane
- Fatty Acids, Omega-3
- Female
- Geriatric Assessment
- Humans
- Lymphocyte Subsets
- Phospholipids
- Severity of Illness Index
The immune system changes with age. In this study we characterized immune changes by performing immunologic screening profiles on ageing individuals. This study was performed at Akdeniz University, in the Faculty of Medicine, Department of Immunology. Healthy volunteers consisted of a younger group (22 donors) and an older group (45 individuals). All subjects had no serious health problems (i.e. chronic heart, lung, liver or immunological diseases) and were taking no prescribed medications. Flow cytometry analysis was used to evaluate CD3, CD4, CD8, CD16, CD19, CD28, CD40, CD45, CD56, CD80, CD86, CTLA-4 and ELISA for IL-1 beta, IL-2, IL-6, IL-10, IFN-gamma, TNF-alpha expression In addition, NK activity and induced cytokine expression (by bioassay and ELISA, respectively) were evaluated. No statistical differences were observed between the two groups in expression of CD3, CD8, CD19, CD80, CD86, CD16, CD 56, or CD28. A higher frequency of expression of CD4, CTLA-4, CD40, and CD45 was seen in older subjects by comparison with younger subjects. Cytokine profiles expressed by stimulated monocytes and lymphocytes from the two groups showed no difference in IL-1 beta, IL-2, IL-6, IL-10, TNF-alpha, and IFN-gamma production levels. We found increased expression levels of CD40 and CD45 levels in healthy older (age: 59.42 /- 5.89) versus younger individuals (age: 30.32 /- 2.29). CTLA-4 expression levels were also higher in older subjects, with no difference in CD28 expression levels between younger/older individuals.
MeSH Terms
- Adult
- Age Factors
- Aging
- Biomarkers
- CD40 Antigens
- CTLA-4 Antigen
- Cytokines
- Female
- Flow Cytometry
- Humans
- Immunity, Humoral
- Leukocyte Common Antigens
- Lymphocytes
- Male
- Middle Aged
- Monocytes
The aim of the present study was to determine Toll-like receptor (TLR)-2 and TLR-4 membrane expression on the major peritoneal leukocyte populations throughout the aging process, including subjects that had achieved exceptional longevity. ICR (CD1) female mice of different ages: adult (44 /- 4 weeks), old (69 /- 4), very old (92 /- 4) and extreme long-lived (125 /- 4), were used. Peritoneal leukocytes were collected, and percentages of CD11b, CD11c, CD3CD4, CD3CD8 and CD19 cells present in the samples were analysed, as well as the expression of TLR-2 and TLR-4 on them, by flow cytometry. The results showed increased TLR expression on CD11b cells from animals at very old ages and especially in the extreme long-lived. Old subjects showed lower percentage of CD11c cells, but no age-related changes were found in the TLR expression on these cells. TLRs on CD3CD4 and CD3CD8 cells from very old animals were increased as compared to the adults, whereas long-lived subjects showed preserved levels. However, TLR expression on CD19 cells was higher in long-lived individuals with respect to subjects at all the other younger ages. These data suggest that differential age-related changes in the expression of TLR-2 and TLR-4 on leukocyte populations from long-lived and non-selected younger old mice could contribute to a different age-related immune remodelling in long-lived subjects, which could allow better preservation of their immune responses.
MeSH Terms
- Age Factors
- Aging
- Animals
- Antigens, CD
- Female
- Humans
- Leukocytes
- Life Expectancy
- Longevity
- Mice
- Peritoneum
- Toll-Like Receptor 2
- Toll-Like Receptor 4
The number of circulating B-cells in peripheral blood plateaus between 2 and 24 months of age, and thereafter declines gradually. How this reflects the kinetics of the precursor B-cell pool in the bone marrow is of clinical interest, but has not been studied thoroughly in humans. The authors analyzed bone marrow (n = 37) from healthy children and adults (flow cytometry) searching for age-related changes in the total precursor B-cell compartment. In an age-matched cohort (n = 25) they examined age-related global gene expression changes (Affymetrix) in unsorted bone marrow with special reference to the recombination activating gene 1, RAG1. Subsequently, they searched the entire gene set for transcripts correlating to the RAG1 profile to discover other known and possibly new precursor B-cell related transcripts. Both methods disclosed a marked, transient increase of total precursor B-cells at 6-20 months, followed by a rapid decrease confined to the first 2 years. The decline thereafter was considerably slower, but continued until adulthood. The relative composition of total precursor B-cells, however, did not change significantly with age. The authors identified 54 genes that were highly correlated to the RAG1 profile (r >or= .9, p < 1 x 10(-8)). Of these 54 genes, 15 were characteristically B-lineage associated like CD19, CD79, VPREB, EBF1, and PAX5; the remaining 39 previously not described as distinctively B-lineage related. The marked, transient increase in precursor B-cells and RAG1 transcriptional activity is not reflected by a similar peak in B-cells in peripheral blood, whereas the sustained plateau concurs in time.
MeSH Terms
- Adolescent
- Adult
- Aging
- B-Lymphocyte Subsets
- Bone Marrow
- Bone Marrow Examination
- Cell Lineage
- Child
- Child, Preschool
- Cohort Studies
- Female
- Flow Cytometry
- Gene Expression Profiling
- Gene Expression Regulation, Developmental
- Hematopoietic Stem Cells
- Homeodomain Proteins
- Humans
- Infant
- Infant, Newborn
- Lymphocyte Count
- Male
- RNA, Messenger
- Transcription, Genetic
- Young Adult
Leucine-rich repeat kinase 2 (LRRK2) is the causal molecule of familial Parkinson's disease (PD), but its true physiological function remains unknown. In the normal mouse, LRRK2 is expressed in kidney, spleen, and lung at much higher levels than in brain, suggesting that LRRK2 may play an important role in these organs. Analysis of age-related changes in LRRK2 expression demonstrated that expression in kidney, lung, and various brain regions was constant throughout adult life. On the other hand, expression of both LRRK2 mRNA and protein decreased markedly in spleen in an age-dependent manner. Analysis of purified spleen cells indicated that B lymphocytes were the major population expressing LRRK2, and that T lymphocytes showed no expression. Consistently, the B lymphocyte surface marker CD19 exhibited an age-dependent decrease of mRNA expression in spleen. These results suggest a possibly novel function of LRRK2 in the immune system, especially in B lymphocytes.
MeSH Terms
- Age Factors
- Aging
- Animals
- B-Lymphocytes
- Leucine-Rich Repeat Serine-Threonine Protein Kinase-2
- Mice
- Mice, Inbred C57BL
- Protein-Serine-Threonine Kinases
- RNA, Messenger
- Spleen
The aims of this flow cytometry study were to quantify B lymphoid precursors known as hématogones across age and clinical conditions and to study the immunophenotypic profile of these benign immature B cells. A total of 406 consecutive marrow specimens were analyzed for hématogones using 4-color flow cytometry during a 19 month period (60% males and 40% females). The age range was 3 months to 89 years. Hématogones were present in 80% of the specimens. Morphologic analysis of the smears from each patient showed small numbers of hématogones (<13% of total cellularity). The B cell population was defined by CD19 CD45 bright positivity, coexpression of other B lineage markers: CD20, CD22, CD10, CD29, CD38 and CD58 in addition to HLA-DR and CD34. In our study we found a significant decline in hématogones with increasing age but a broad range was found at all ages. Marrow from some adults contained relatively high numbers. Diagnosis in these patients included cytopenias, infections, and neoplastic diseases. Distinction of hématogones is critical for disease management particularly after therapy of paediatric B acute lymphoblastic leukaemia to monitor for minimal residual disease.
MeSH Terms
- Adolescent
- Adult
- Aged
- Aged, 80 and over
- Aging
- Antigens, CD
- Child
- Child, Preschool
- Female
- Flow Cytometry
- Humans
- Immunophenotyping
- Infant
- Male
- Middle Aged
- Precursor Cells, B-Lymphoid
- Prospective Studies
- Young Adult
Due to the fact that the coexpression of CD23 and CD27 has been reported to occur in B lymphocytic leukaemic clones and that there is debate about CD23 expression on memory B cells, we evaluated the behaviour of naive B cells (CD23-/CD27-) and memory B cells (CD27 ) in the peripheral blood of a large number of humans of all ages. B cells were also distinguished into B2 (CD5-) and B1-a cells (CD5 ). The cell surface expression of CD19, CD5, CD23 and CD27 was assessed on peripheral blood lymphocytes from 1,427 subjects of all ages undergoing peripheral blood immunophenotyping for a variety of reasons. The absolute number of B lymphocytes and the percentage of naive cells (CD23-/CD27-) decreased with age whereas there was an increase in memory cells (CD27 ). A small subset of B cells co-expressing CD23 and CD27 was present in humans of all ages, although the majority of CD27 cells were CD23-. The percentages and rate of increase with age of B1-a CD23 /CD27 were slightly higher than those of B2 cell counterparts. On the basis of our data, age-associated changes in surface markers of B cells seem to be finely balanced and probably related to functional changes after antigen encounters, while the whole peripheral blood B-cell compartment undergoes a quantitative regression.
MeSH Terms
- Aging
- Antigens, CD19
- B-Lymphocyte Subsets
- CD5 Antigens
- Female
- Gene Expression
- Humans
- Male
- Receptors, IgE
- Tumor Necrosis Factor Receptor Superfamily, Member 7
Keywords
- B lymphocytes
- B-1a cells
- CD23
- CD27
The purpose of this study was to evaluate the inflammatory cell subset proportions in the upper gingival connective tissue, including mature dendritic cells (DC) in elderly and younger patients with generalized chronic periodontitis in order to further understand the effect of aging on gingival inflammatory phenomenon. Gingival tissue specimens presenting chronic periodontitis from 8 elderly patients aged >75 (test group, group T) and from 8 younger patients aged 50-60 (considered as controls, group C) were analysed by immunohistochemistry using monoclonal antibodies against CD45RB, CD4, CD8, CD19, CD68, DC-SIGN, DC-LAMP molecules. The number of each immunolabelled cells subset was counted using image analysis. The difference in the number of CD45RB leucocytes in the upper gingival connective tissue between groups was not significant permitting to use it as reference. As compared to group C, the lymphocyte subsets/CD45RB leucocytes ratios tended to decrease in group T but the decrease was significant only for CD4 T lymphocytes/CD45RB cells ratio (p<0.03). On the opposite, the ratios of antigen-presenting cells DC-SIGN cells/CD45RB cells and DC-LAMP cells/CD45RB cells were significantly increased (p<0.03 and <0.0001, respectively) in group T. Moreover, in group T the DC-LAMP cells/DC-SIGN cells ratio was significantly increased (p<0.05) showing an increased number of matured dendritic cells. During chronic periodontitis in elderly patients, our results show a decrease in the ratio of gingival CD4 lymphocyte subset associated with an increase in the ratios of antigen-presenting cells subsets and more particularly maturated DC-LAMP dendritic cells.
MeSH Terms
- Aged
- Aging
- CD4-CD8 Ratio
- CD4-Positive T-Lymphocytes
- CD8-Positive T-Lymphocytes
- Chronic Periodontitis
- Dendritic Cells
- Female
- Gingiva
- Humans
- Immunohistochemistry
- Male
- Middle Aged
- T-Lymphocyte Subsets
To determine whether exercise increases endothelial progenitor cells (EPCs) in patients with peripheral vascular disease, we developed a multi-parameter flow cytometry assay to rigorously assess EPCs and mature endothelial cells (ECs) in control subjects and patients with peripheral artery disease (PAD) subjected to graded exercise. Blood was collected from young healthy subjects (n = 9, mean age 33 years), older healthy subjects (n = 13, mean age 66 years), and older subjects with PAD (n = 15, mean age 69 years) before and 10 minutes after exercise. White blood cells were isolated and stained with a five-color antibody panel: FITC-anti-CD31, PE-anti-CD146, PE-anti-CD133, PerCP-Cy5.5-anti-CD3,-CD19,-CD33, PE-Cy7-anti-CD34, and APC-anti-VEGF-R2. Viability was assessed by propidium iodide exclusion. Viable, low, side scatter singlets that were CD3-, 19-, and 33-negative were counted. While baseline levels of EPCs and ECs were similar among all subjects, young healthy subjects demonstrated significantly greater (p < 0.05) levels of progenitor cells (PCs) than older healthy and PAD subjects. Levels of EPCs and ECs tended to increase in all subjects after exercise; however, increases in PCs were only observed in young healthy and PAD subjects. Further, trends in the magnitude of change of subsets with exercise were most similar between young and PAD subjects. Our findings suggest that aging may reduce baseline circulating levels of PCs, but not EPCs or ECs, and that exercise-induced mobilization of subsets may differ depending on age and presence of PAD.
MeSH Terms
- Adult
- Aged
- Aging
- Antigens, CD
- Cell Differentiation
- Cell Lineage
- Cell Shape
- Cells, Cultured
- Colony-Forming Units Assay
- Endothelial Cells
- Exercise
- Flow Cytometry
- Humans
- Intermittent Claudication
- Lectins
- Lipoproteins, LDL
- Peripheral Vascular Diseases
- Stem Cells
- Vascular Endothelial Growth Factor Receptor-2
- von Willebrand Factor
Age and cardiovascular disease status appear to alter numbers and function of circulating endothelial progenitor cells (EPCs). Despite no universal phenotypic definition, numerous studies have implicated progenitors with apparent endothelial potential in local responses to vascular injury and with cardiovascular disease in general. To further define the role of this lineage in peripheral artery disease (PAD), we developed a multiparameter flow cytometry assay to analyze multiple phenotypic definitions of progenitor cells (PCs), EPCs, and mature endothelial cells (ECs) and evaluate effects of age and PAD on baseline levels of each subset. Blood was collected from young healthy subjects (N = 9, mean age 33 /- 8 years), older healthy subjects (N = 13, mean age 66 /- 8 years), and older subjects with PAD (N = 15, mean age 69 /- 8 years). After ammonium chloride lysis, cells were stained and analyzed on a Becton-Dickinson LSR II with a 5-color antibody panel: FITC-anti-CD31, PE-anti-CD146, PE-anti-CD133, PerCP-Cy5.5-anti-CD3,-CD19,-CD33 (lineage panel), PE-Cy7-anti-CD34, and APC-anti-VEGF-R2. Viability was assessed by propidium iodide exclusion, and only viable, low to medium side scatter lineage-negative singlets were analyzed. In some studies, cells were sorted for morphological studies. Subsets were defined as indicated later. Our results, using a comprehensive flow cytometric panel, indicate that CD133 , CD34 , and CD133 /CD34 PCs are elevated in younger healthy individuals compared to older individuals, both healthy and with PAD. However, the number of EPCs and mature ECs did not significantly differ among the three groups. Assessment of endothelial colony forming units and dual acLDL-lectin staining supported the flow cytometric findings. We describe a comprehensive flow cytometric method to detect circulating mature and progenitor endothelial populations confirmed by conventional morphological and functional assays. Our findings suggest that aging may influence circulating levels of PCs, but not EPCs or ECs; PAD had no effect on baseline levels of any populations investigated. This study provides the basis for evaluating the potential effects of acute stress and therapeutic intervention on circulating progenitor and endothelial populations as a biomarker for cardiovascular status.
MeSH Terms
- AC133 Antigen
- Adult
- Aged
- Aging
- Antigens, CD
- Antigens, CD34
- Biomarkers
- Cell Count
- Colony-Forming Units Assay
- Endothelial Cells
- Flow Cytometry
- Glycoproteins
- Humans
- Middle Aged
- Peptides
- Peripheral Vascular Diseases
- Phenotype
- Stem Cells
Disturbances in immunity and nutrition status worsen in peritoneal dialysis (PD) patients with advancing age. In the present study, we evaluated variations in total lymphocyte count (TLC) and subset lymphocyte counts (SLCs) with respect to the age of PD patients. We carried out the study in two groups of PD patients. Group I patients (n = 12) were less than 40 years of age (35.5 /- 5.4 years), and their PD duration was 18.2 /- 9.4 months. Group II patients (n = 14) were more than 60 years of age (67.2 /- 5.1 years), and their PD duration was 20.6 /- 11.0 months. In group I, 9 patients were taking angiotensin converting enzyme inhibitors (ACEIs); in group II, 10 patients were taking ACEIs. We used flow cytometry to estimate SLCs (determining CD3, CD4, CD8, CD19, and CD16 56 antigens). In both groups, the mean CD19, CD4, and CD8 counts were lower than the normal ranges. In group II, TLC and CD3 count were also lower than normal. In group I, correlations were seen between age and TLC, CD3, CD19, CD4, and CD8. Correlations were also seen between dialysis duration and TLC, CD3, CD19, and CD4, and between total ACEI dose and CD19 count. In group II, correlations were seen between age and TLC, CD3, and CD8. No correlation was observed between PD duration and TLC or SLCs, but a correlation between total ACEI dose and CD8 count was seen. In patients who were taking enalapril as their only ACEI, a correlation was observed between total enalapril dose and TLC, CD3, and CD8. Our results confirm data that indicate worse immunity and nutrition status in older PD patients and demonstrate decreasing values of TLC and SLCs with aging in younger and older PD patients alike. Administration of ACEIs negatively influences SLCs independently of age, but decreases in TLC and SLCs are significantly related to PD duration only in younger patients.
MeSH Terms
- Adult
- Age Factors
- Aged
- Aging
- Angiotensin-Converting Enzyme Inhibitors
- Antigens, CD
- Female
- Humans
- Lymphocyte Count
- Lymphocyte Subsets
- Male
- Middle Aged
- Nutritional Status
- Peritoneal Dialysis
- Time Factors
Peripheral blood lymphocyte subsets need to be determined in a large, urban, minority-predominant cohort of healthy children to serve as suitable control subjects for the interpretation of the appearance of these cells in several disease conditions, notably pediatric HIV-1 infection. We sought to determine the distribution of lymphocyte subsets in healthy urban-dwelling infants, children, and adolescents in the United States. Lymphocyte subsets were determined by means of 3-color flow cytometry in a cross-sectional study of 807 HIV-unexposed children from birth through 18 years of age. Cell-surface marker analysis demonstrated that age was an extremely important variable in 24 lymphocyte subset distributions measured as percentages or absolute counts--eg, the CD4 (helper) T cell, CD8 (cytotoxic) T cell, CD19 B cell, CD4CD45RACD62L (naive helper) T cell, CD3CD4CD45RO (memory helper) T cell, CD8HLA-DRCD38 (activated cytotoxic) T cell, and CD8CD28 (activation primed cytotoxic) T cell. The testing laboratory proved to be an important variable, indicating the need for using the same laboratory or group of laboratories to assay an individual's blood over time and to assay control and ill or treated populations. Sex and race-ethnicity were much less important. The results of this study provide a control population for assessment of the effects of HIV infection on the normal development and distribution of lymphocyte subsets in children of both sexes, all races, and all ethnic backgrounds from birth through 18 years of age in an urban population. This study's findings will also prove invaluable in interpreting the immune changes in children with many other chronic diseases, such as primary immunodeficiency, malignancy, rheumatoid arthritis, and asthma.
MeSH Terms
- Adolescent
- Aging
- Child
- Child, Preschool
- Continental Population Groups
- Cross-Sectional Studies
- Ethnic Groups
- Female
- Flow Cytometry
- Humans
- Infant
- Infant, Newborn
- Lymphocyte Count
- Lymphocyte Subsets
- Male
- Reference Values
- Regression Analysis
- United States
- Urban Population
Keywords Non-programmatic
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
Comparisons of lymphocyte subsets show that the ratio of CD4 to CD8 is usually greater than one. Inversion of this ratio was found to predict survival in a Swedish octogenarian sample (n=27 deaths), although individual lymphocyte subsets did not predict survival. We have examined these relationships in a larger sample (n=153 deaths). Inversion of the CD4 to CD8 ratio was present in 16% of the sample and predicted survival when adjusted for age but not when adjusted for sex. For individual lymphocyte subsets, higher CD4 and CD19 percentages were associated with better survival, but only the CD19 percentage remained significant when adjusted for age and sex.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- CD4-CD8 Ratio
- Cluster Analysis
- Female
- Humans
- Lymphocyte Subsets
- Male
- Mortality
- Predictive Value of Tests
- Survival Rate
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
The immune status is a parameter of the capacity of the immune system to fend off microorganisms. The use of leukocyte subtyping to define the immune status is clinically established. IFN-gamma is a key cytokine directing the immune response. In this study, we investigated whether IFN-gamma is a more sensitive parameter of the immune status. All persons tested showed stable quantities of white blood cell counts over the whole study. Analyses of the lymphocyte subpopulations of two time points resulted in a strong correlation with high statistical significance for the percentage of CD3, CD4, CD8, CD19, CD45 -subtypes and CD56/16 positive cells. IFN-gamma production by the individuals correlates between these time points also, but only if an ELISA strongly correlating to IFN-gamma bioactivity was used instead of other commercial IFN-gamma ELISAs. The IFN-gamma production by males was less variable than by females. Furthermore, intraindividual differences in IFN-gamma secretion were minimal after the age of 46. In conclusion, our data demonstrate that IFN-gamma is a more sensitive parameter for the actual status of the immune system, since its titer shows alterations faster than leukocyte subtyping. Furthermore, leukocyte subtypes do not correspond to the production capacity of the regulatory cytokine IFN-gamma in healthy individuals.
MeSH Terms
- Adult
- Aging
- Antigens, CD
- Enzyme-Linked Immunosorbent Assay
- Female
- Humans
- Immune System
- Interferon-gamma
- Lymphocyte Count
- Male
- Middle Aged
- Sex Characteristics
- Time Factors
The aim of the present study was to determine the effect of repeated tonsillitis on the development of lymphocyte subsets in the tonsils and among peripheral blood lymphocytes (PBL) of children. Subsets of T- and B cells were analyzed in the tonsils and in PBL of patients undergoing tonsillectomy for idiopathic tonsillar hypertrophy, recurrent tonsillitis, or tonsillar hypertrophy and tonsillitis. The majority of the CD4 cells in the tonsils displayed the CD45RO phenotype, while the majority of those in the PBL displayed the CD45RA phenotype. Likewise, the proportion of CD45RO CD8 cells was higher in the tonsils than among PBL. The proportion of CD4 cells expressing the CD45RO marker increased with age among PBL, but not in the tonsils. B cells, detected by their CD19, CD20, and CD21 markers, were three times more abundant in the tonsils than in the PBL. The proportion of CD38 cells showed a negative correlation with age, both in the tonsils and among PBL. Among PBL a striking age-related reduction was seen in the proportion of CD19 , CD21 and CD38 CD21 B cells. In contrast, in the tonsils age-related changes could be detected only in the proportion of CD21 CD38 cells. No difference among patients with various clinical diagnoses was detectable in any of the T- and B cell subsets in the tonsils and PBL. Thus, lymphocyte subsets evolve independently in the tonsils and peripheral blood, with the repeated antigenic challenge of tonsillar lymphocytes not influencing circulating memory cells.
MeSH Terms
- ADP-ribosyl Cyclase
- ADP-ribosyl Cyclase 1
- Adolescent
- Aging
- Antigens, CD
- Antigens, Differentiation
- B-Lymphocyte Subsets
- CD4-Positive T-Lymphocytes
- Child
- Child, Preschool
- Flow Cytometry
- Fluorescent Antibody Technique
- Humans
- Immunologic Memory
- Infections
- Leukocyte Common Antigens
- Lymphocyte Subsets
- Membrane Glycoproteins
- NAD Nucleosidase
- Palatine Tonsil
- Receptors, Complement 3d
- T-Lymphocyte Subsets
- Tonsillitis
Immune status was determined in a representative sample of elderly people by measuring lymphocyte subsets in whole-blood samples as part of an epidemiological study of the population aged 65 and over. Venepuncture was undertaken in more than 500 individuals who took part in an extensive interview that focused on the lifestyle and psychosocial determinants of healthy aging. The results show that median levels of all lymphocyte subsets tend to decline as the age of the sample increases. In the total sample there were significant age effects (p < 0.05) on total lymphocytes, CD3, CD4, and CD19 (B cells); age differences did not reach significance for CD8 and CD57. There were also significant sex differences (p < 0.05) on CD3, CD4, and CD19, and in all cases women had higher values than men. When we selected a particularly healthy subsample who did not report any illness and took no medication, the findings were unchanged. We conclude that the peripheral expression of lymphocytes appears little affected by aging-related illnesses in the general population, but is affected by aging itself. The study provides reference values for the lymphocyte measures, which can be regarded as having greater validity than the values usually cited.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- CD4-CD8 Ratio
- Female
- Humans
- Lymphocyte Count
- Lymphocyte Subsets
- Male
A randomized double-blind trial was performed in order to assess the efficacity of differing combinations of antioxidant nutrients on biochemical parameters of vitamin and trace element status, immunological parameters and free radical metabolism in elderly long term hospitalized subjects. A total of 756 institutionalized elderly subjects were recruited in 26 nursing homes in different areas of France. Four groups were constituted, receiving daily, for 1 year, either vitamins (beta-carotene, 6 mg; vitamin C, 120 mg; and vitamin E, 15 mg), trace elements (zinc, 20 mg and selenium, 100 micrograms), trace elements associated with vitamins, or a placebo. Biochemical indicators of trace elements and vitamin status and free radical parameters were measured before and after 6 months and 1 year of supplementation. Some immunological markers were investigated initially and after 6 months of supplementation on a subsample of 134 subjects. Mean plasma levels of alpha-tocopherol, gamma-tocopherol, vitamin C, alpha-carotene, beta-carotene and copper increased significantly after 6 months of supplementation in groups receiving vitamins alone or associated with trace elements. Serum selenium concentrations were significantly increased at 6 months of supplementation, and serum zinc only after one year in the trace element groups. Serum lycopene levels were significantly decreased by trace element supplementation. A significant increase in Se-glutathione peroxidase (GPx) levels was observed in groups receiving trace elements alone or associated with vitamins. No effect was noted on superoxide dismutase (SOD) activity or TBARs production. No effect of supplementation was found for in vitro lymphocyte proliferative responses or most lymphocyte subsets, except for a significantly lower percentage of CD2 subsets observed in groups receiving mineral supplementation either alone or associated with vitamins. A significant difference in CD19 subsets was found in groups receiving trace elements. Mean IL-1 production was significantly higher after 6 months of supplementation in the vitamin groups.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Double-Blind Method
- Female
- France
- Free Radical Scavengers
- Glutathione
- Hospitalization
- Humans
- Immunity
- Male
- Nursing Homes
- Nutritional Status
- Trace Elements
- Vitamins
The neonatal immune system responds to a restricted range of antigens, producing largely IgM antibody of low affinity. Comparison of the components of the B-cell antigen receptor complex shows significantly elevated membrane levels of IgM in neonatal B cells, compared with adult cells. CD79, which acts as the signal transducer for membrane immunoglobulin, is elevated in parallel with IgM, while IgD is elevated to a lesser degree. CD19, CD21, CD22 and CD81, which are all involved in transmitting activation signals when immunoglobulin is engaged, are not elevated. CD32, which is involved in negative regulation of activation, is present at reduced levels on cord B cells. The elevation of B-cell membrane IgM persists during infancy. Neonatal B cells respond in vitro to interleukin-4 (IL-4) by further elevation of membrane IgM levels. The elevated level of membrane IgM may make neonatal B cells easier to trigger by low concentrations of antigen, but in vitro activation and immunoglobulin modulation experiments did not show significant differences between cord and adult B-cell responses to anti-IgM.
MeSH Terms
- Adult
- Aging
- B-Lymphocyte Subsets
- B-Lymphocytes
- CD5 Antigens
- Cell Culture Techniques
- Fetal Blood
- Humans
- Immunoglobulin M
- Immunologic Capping
- Infant, Newborn
- Interleukin-4
- Receptors, Antigen, B-Cell
Dendritic cells are antigen-presenting cells (APC), which are crucial for the initiation of an immune response. In spite of the well known decline of immune function in old age, no information is yet available on whether dendritic cells are also affected by the ageing process. It was therefore the aim of this study to compare peripheral blood dendritic cells (DC) from old and young healthy individuals. Using granulocyte-macrophage colony-stimulating factor (GM-CSF) and IL-4, DC were propagated from peripheral blood mononuclear cells (PBMC). The obtained cell populations had a typical dendritic morphology and expressed HLA class I and class II, CD23, CD32, CD40, CD44 and CD54, but not CD3 and CD19. Larger numbers of DC were obtained from old individuals than from young ones in spite of a similar expression pattern of surface molecules. DC from aged persons also survived better under in vitro culture conditions. When tested for their antigen-presenting capacity, DC from young and old individuals were equally effective in inducing the proliferation of tetanus toxoid-specific T cell clones after antigenic stimulation. Peripheral blood DC from aged individuals may thus still function as powerful APC. They may represent useful tools for immunotherapy in the aged.
MeSH Terms
- Adolescent
- Adult
- Aged
- Aging
- Antigen Presentation
- Biomarkers
- Cell Division
- Dendritic Cells
- Humans
The paper reports the distribution of lymphocyte subpopulations in the Caucasian population of Romania. Investigations were carried out in cells bearing the following antigens: CD3 (T cells), CD19 (B cells), CD4 (T helper/inducer cells), CD8 (T suppressor/cytotoxic and some NK cells), and CD16 CD56 (NK cells). Reference values for the lymphocyte subpopulation were obtained from over 100 healthy Caucasian adult volunteers. Blood from these donors was analyzed using FACScan flow cytometer, Leuco-GATE, Simultest and FACS Lysing Solution, and SimulSET software. As an internal quality control, it was verified that %T %B %NK approximates 100% in all samples. The results presented here, obtained on healthy donors (51 males and 49 females), showed that there are no statistically significant variations of CD4/CD8 ratio related to sex or age, the mean value of this ratio (2.0 /- 0.02) being similar to that reported by the West-European countries. Additional similarities were found when the relative percentage, mean /- standard deviation (CD3 = 74.2 /- 2.8; CD19 = 10.8 /- 1.6; CD4 = 42.0 /- 2.5; CD8 = 28.9 /- 5.7; CD56 = 15.2 /- 0.4) and the absolute cell number of the major peripheral blood mononuclear subsets established in our study were compared with other published results. This study was entirely supported by Becton Dickinson--Europe (Division Heidelberg, Germany).
MeSH Terms
- Adolescent
- Adult
- Aged
- Aging
- Antigens, CD
- Antigens, Surface
- European Continental Ancestry Group
- Female
- Flow Cytometry
- Humans
- Lymphocyte Count
- Lymphocyte Subsets
- Male
- Middle Aged
- Quality Control
- Reference Values
- Romania
- Sex Characteristics
To determine whether synaptic contact is required to express adult-type nicotinic acetylcholine receptors (A-AChR) in developing mammalian muscle, we have examined single-channel AChR activity in primary muscle cultures maintained for up to 29 days. A-AChRs were first expressed after day 12 in culture (CD12), during a period characterized by the accumulation of embryonic acetylcholine receptors (E-AChR). The highest rate of A-AChR expression was observed between CD15 and 19, during a period of maximal E-AChR accumulation. Although the level of A-AChR expression between individual patches was quite variable during this period, A-AChRs accounted for up to 40% of the events produced by receptors expressed over a 3-day interval. Between CD19 and 29, the density of E-AChRs diminished while the expression of A-AChRs per patch continued to increase but at a lower rate than that observed between CD15 and 19. In 25-29-day cultures, 70.6% of patches exhibited both E-AChR and A-AChR activity, and the percentage of A-AChR events per patch ranged between 0 and 47% with a mean of 11.7 /- 3.2%. These results demonstrate that endogenous muscle mechanisms promote developmental increases in the expression of A-AChRs in myotubes that have no history of synaptic contact. This conclusion suggests that synaptic imprinting at developing junctions is mediated in part by endogenous muscle mechanisms, and does not require direct neurotrophic activation of epsilon mRNA transcription.
MeSH Terms
- Aging
- Animals
- Culture Techniques
- Electrophysiology
- Mice
- Mice, Inbred C57BL
- Muscle Development
- Muscles
- Receptors, Nicotinic
- Synapses
- Time Factors
Peripheral blood lymphocyte subset reference ranges were examined in a large group (N = 130) of healthy pediatric patients ranging in age from 1 month to 17 years. All samples were stained with monoclonal antibodies, processed with a whole blood lysis technique, and analyzed on a flow cytometer. Data analysis demonstrated statistically significant changes in most lymphocyte subsets at age 3 years. The relative and absolute numbers of total lymphocytes, CD2, CD4, and CD19 cells; absolute numbers of CD3 and CD8 cells; and CD4/CD8 ratios were high at birth, decreased during early childhood, and closely approximated adult reference values after age 3 years. The relative numbers of CD8 lymphocytes were low in early childhood and then rose to adult values after 3 years of age. The relative percentage of CD3 cells remained stable over all ages studied. Although "adult" lymphocyte subset reference ranges may be similar to those in children older than 3 years, age-adjusted reference ranges should be used for the early childhood period.
MeSH Terms
- Adolescent
- Aging
- Antigens, CD
- Child
- Child, Preschool
- Humans
- Infant, Newborn
- Lymphocyte Subsets
- Lymphocytes
- Reference Values
As a part of an ongoing longitudinal investigation, this study examined relationships between survival and selected immune system parameters in a sample (n = 102) of very old individuals (86-92 years at the time of initial immune system data collection). Analyses were performed comparing initial time-point measurements from those individuals who were alive (n = 75) and those who were deceased (n = 27) two years after initial data collection. Immune system measurements consisted of determination of peripheral blood lymphocytes and lymphocyte subsets, as well as T-cell responses to activation by Concanavalin A. Cluster analysis identified a subgroup associated with nonsurvival which indicated characteristics that included: poor T-cell proliferative responses, high CD8 percentages, and low CD4 and CD19 percentages. This multivariate analysis suggested that combinations of immune system parameters predict two-year survival otherwise not apparent when single immune system parameters were evaluated in the elderly.
MeSH Terms
- Aged
- Aged, 80 and over
- Aging
- Antigens, CD
- Cell Division
- Cluster Analysis
- Concanavalin A
- Humans
- Immune System
- Longitudinal Studies
- Lymphocyte Count
- Lymphocyte Subsets
- Retrospective Studies
- Survival Analysis
- Survivors
- Sweden
- T-Lymphocytes
Peripheral blood mononuclear cells were quantified for the subsets of CD4, CD8, and CD19 lymphocytes by using CD45RA (2H4), CD29(4B4), CD57, CD5, CD10, Leu8, HLA-DR, and TCR gamma delta-1 monoclonal antibodies and dual color immunofluorescence. A comparative analysis of lymphocyte subpopulations was made among 52 HIV-infected and 50 age-matched control children and 30 HIV-seropositive and 27 negative control adults. A significant decrease in the CD4 CD45RA "naive" cells was much more marked in HIV-infected children than in HIV-infected adults. A significant percentage increase in the CD4 CD29 "memory" cells was observed in HIV-infected children but not in infected adults; however, the absolute numbers were usually decreased in all age groups. The mean percentage and absolute numbers of CD4 CD7 and CD4 Leu8 cells were decreased in HIV-infected children, although usually not significantly. The CD3 TCR gamma delta-1 did not show any change in the infected children tested. The mean percentage and absolute number of the CD8 HLA-DR cells increased significantly in HIV-infected persons of all ages. The CD8 CD57 cells were increased in percentage and absolute number in HIV-infected children ages 1-4 and 4-8 years. In the adults, no change was noted in either the percentage or absolute number of CD19 CD5 B cells, a finding similar to that noted in HIV-infected children above 1 year of age. Although adults showed a significant decrease in both percentage and numbers of CD5- B cells, an increase was noted in the 7- to 12-month-old HIV-infected children. The CD19 CD10 cells showed a slight but significant decrease in the youngest age group and a significant increase in the older age groups of HIV-infected children. These findings indicate that several lymphocyte subpopulations are altered differentially during HIV infection in children of varying ages and in adults.
MeSH Terms
- Adult
- Aging
- Antigens, CD
- Antigens, CD19
- Antigens, Differentiation, B-Lymphocyte
- B-Lymphocyte Subsets
- CD4-Positive T-Lymphocytes
- CD8 Antigens
- Child
- Child, Preschool
- HIV Infections
- Humans
- Immunologic Memory
- Infant
- Integrin beta1
- Neprilysin
- T-Lymphocyte Subsets
CD19 is a B-cell-specific member of the immunoglobulin superfamily expressed from early pre-B-cell development until plasma cell differentiation. In vitro studies demonstrate that the CD19 signal transduction molecule can serve as a costimulatory molecule for activation through other B-lymphocyte cell surface molecules. However, much remains to be known regarding how CD19 functions in vivo and whether CD19 has different roles at particular stages of B-cell differentiation. Therefore, transgenic mice overexpressing the human CD19 (hCD19) gene were generated to determine whether this transgene would be expressed in a B-lineage-specific fashion and to dissect the in vivo role of CD19 in B-cell development and activation. Expression of the human transgene product was specifically restricted to all B-lineage cells and appeared early in development as occurs with hCD19. In addition, expression of hCD19 severely impaired the development of immature B cells in the bone marrow, with dramatically fewer B cells found in the spleen, peripheral circulation, and peritoneal cavity. The level of hCD19 expressed on the cell surface correlated directly with the severity of the defect in different transgenic lines. These results demonstrate that the hCD19 gene is expressed in a lineage-specific fashion in mice, indicating that the hCD19 gene may be useful for mediating B-lineage-specific expression of other transgene products. In addition, these results indicate an important role for the lineage-specific CD19 molecule during early B-cell development before antigen-dependent activation.
MeSH Terms
- Aging
- Animals
- Antibodies, Monoclonal
- Antigens, CD
- Antigens, CD19
- Antigens, Differentiation, B-Lymphocyte
- B-Lymphocytes
- Bone Marrow
- Crosses, Genetic
- Female
- Flow Cytometry
- Genetic Carrier Screening
- Homozygote
- Humans
- Immunoglobulin M
- Immunohistochemistry
- Lymph Nodes
- Male
- Mice
- Mice, Inbred C57BL
- Mice, Transgenic
- Organ Specificity
- Restriction Mapping
- Signal Transduction
- Spleen
Children with ALL diagnosed at less than 2 years of age have a poor prognosis when compared with older children. In an effort to identify biologic features of ALL in children less than 2 that might explain this difference, we performed extensive immunophenotypic and molecular genetic analyses on a series of patients. For comparison purposes patients were divided into four groups: CALLA- (CD10-) infants less than 2 years of age at diagnosis (n = 10), CALLA- children greater than 2 years of age at diagnosis (n = 10), CALLA infants (less than 2 years, n = 21) and CALLA children (older than 2 years, n = 21). No immunophenotyping differences in CALLA- or CALLA subgroups were identified when cases less than 2 were compared with cases greater than 2 years of age at diagnosis. The most interesting results were in the CALLA- group where 94% of the samples expressed the B cell antigen CD19 but 27% co-expressed CD7. Double labeling experiments confirmed leukemic blast cells co-expressed CD19 and CD7. The double-labeled cells represent either leukemic conversion of a precursor cell which has not yet committed to B or T cell lineage or aberrant expression of these antigens. Molecular genetic studies demonstrated that all samples, regardless of the patients' age or immunophenotype, had rearrangement of the Ig heavy chain gene. The most striking molecular results were in CALLA- patients; in patients less than 2 at diagnosis neither the beta- nor the gamma-chain gene of the T cell receptor (TCR) was rearranged, whereas DNA from 5 of 10 patients over the age of 2 demonstrated beta- or gamma-chain TCR gene rearrangements. The percentage of CALLA cases under the age of 2 years with rearrangements in TCR genes is less than that found in CALLA cases over the age of 2 years. The finding of no TCR rearrangements in CALLA- ALL and a decreased number of gamma-TCR rearrangements in CALLA cases under the age of 2 suggest that age may affect TCR gene rearrangements in lymphoblasts. The molecular differences in TCR gene rearrangements do not appear to correlate with the response to therapy.
MeSH Terms
- Aging
- Antigens, Differentiation
- Antigens, Differentiation, B-Lymphocyte
- Antigens, Neoplasm
- B-Lymphocytes
- Biomarkers, Tumor
- Burkitt Lymphoma
- Gene Rearrangement, T-Lymphocyte
- Hematopoietic Stem Cells
- Humans
- Infant
- Neprilysin
- Prognosis
- Receptors, Antigen, T-Cell
The circulating lymphocytes of 88 consecutive patients following autologous, conventional, or T-cell depleted bone marrow transplantation were serially analyzed for B-cell surface antigen expression and function. In the majority of patients, except for those who developed chronic graft-versus-host disease, the number of circulating CD20 B cell normalized by the fourth posttransplant month. The earliest detectable B cells normally expressed HLA-DR, CD19, surface immunoglobulin (slg), CD21, Leu-8, and lacked expression of CD10 (CALLA). In addition, the circulating B cells expressed CD1c, CD38, CD5, and CD23 for the first year following transplant, antigens that are normally expressed on a small percentage of circulating B cells in normal adults, but highly expressed on cord blood B cells. Similar to cord blood B cells, patient B cells isolated during the first year following transplant, proliferated normally to Staphylococcus aureus Cowan strain I (SAC), and produced IgM, but minimal or no IgG when stimulated with pokeweed mitogen and SAC, unlike normal adult B cells that produce both. The similar phenotype and function of posttransplant and cord blood B cells, and their similar rate of decline in patients and normal children adds further evidence to support the hypothesis that B-cell differentiation posttransplant is recapitulating normal B-cell ontogeny.
MeSH Terms
- Adult
- Aging
- Antigens, CD
- Antigens, CD20
- Antigens, Differentiation, B-Lymphocyte
- B-Lymphocytes
- Bone Marrow Transplantation
- Cell Differentiation
- Child
- Child, Preschool
- Graft vs Host Disease
- HLA-DR Antigens
- Humans
- Immunoglobulin G
- Immunoglobulin M
- Immunophenotyping
- Infant, Newborn
- Leukocyte Count
- Lymphocyte Activation
- Staphylococcus aureus
- T-Lymphocytes