Relative and absolute numbers of T lymphocyte subpopulations and NK cells in male population in relation to age

Using BMA monoclonal antibodies and fluorescent microscope, percentages and absolute numbers of lymphocytes, T cell subsets and NK cells were enumerated in peripheral blood from 126 healthy men. Although absolute numbers of total lymphocytes did not differ according to age, the numbers and percentage of natural killer (NK) cells showed positive interrelationship with age. The percentage but not absolute numbers of cells reacting with BMA 030 (CD3) and BMA 040 (CD4) antibodies were significantly increased only in groups aged of 20-29 yrs and 30-39 yrs. The percentage and number helper/inducer T cells (CD4) were comparable in the four groups of subjects. These results indicate that peripheral lymphocyte populations and T cell subsets and NK cells remarkably vary in healthy men over a wide range of ages.     

Effects of single dose compared with three days' prednisolone treatment of healthy volunteers: contrasting effects on circulating lymphocyte subsets.

AIMS--To investigate the effects of longer term corticosteroid treatment on circulating lymphocyte subsets. METHODS--Prednisolone (20 mg daily) was given to 12 healthy volunteers in a single morning dose for three days. Circulating lymphocyte subsets were measured by flow cytometry after whole blood lysis. RESULTS--Seven hours after the first dose of prednisolone there was a significant fall in absolute numbers of lymphocytes, T cells, CD4+ and CD8+ cells, and B cells. The percentage of T cells fell significantly, due to a fall in percentage of CD4+ cells. In contrast to the seven hour findings, at 72 hours there was a significant rise in absolute numbers of lymphocytes, T cells, CD4+, CD8+, and B cells. This trend was already apparent by 24 hours. The percentage of CD4+ cells was significantly raised at 72 hours, while that of CD8+ cells had fallen significantly. The percentage of natural killer cells had fallen at 72 hours; that of B cells remained increased at 72 hours. CONCLUSIONS--These findings show that corticosteroid treatment causes significant changes in lymphocyte subsets, and that such changes must be considered when designing studies of lymphocyte subsets during illness.     

Effects of Ageing on the Immune System: Infants to Elderly

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 lympho‐cyte 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.     

Age-Related Changes in Blood Lymphocyte Subsets of Saudi Arabian Healthy Children

The age-related changes in absolute and percentage values of lymphocyte subsets in the peripheral blood of healthy children of different ages (1 month to 13 years) were studied by flow cytometry. The absolute and percentage values for most lymphocyte subpopulations differed substantially with age. Comparisons among age groups from infants through adults revealed progressive declines in the absolute numbers of leukocytes, total lymphocytes, and T, B, and natural killer (NK) cells. The percentages of T cells increased with age. Within the T-lymphocyte population, the CD8⁺ subset increased but the CD4⁺ subset decreased, resulting in a declining CD4⁺/CD8⁺ ratio. The percentage of B cells declined, but that of NK cells remained unchanged. The percentage of HLA-DR⁺ T cells increased over time, but their number changed inconsistently. Our findings confirm and extend earlier reports on age-related changes in lymphocyte subpopulations. These data should be useful in the interpretation of disease-related changes, as well as therapy-dependent alterations, in lymphocyte subsets in children of different age groups.     

The decrease in peripheral blood CD4+ T cells following thermal injury in humans can be accounted for by a concomitant decrease in suppressor-inducer CD4+ T cells as assessed using anti-CD45R

Using single- and two-color fluorescence flow cytometry, 10 thermally injured human subjects were assessed over time for both percentages and absolute numbers of lymphocytes comprising peripheral blood lymphocyte subpopulations. The CD3+ lymphocyte percentage decreased significantly in the early postburn period, and this decrease could be accounted for entirely by a concomitant decrease in the CD4+ lymphocyte percentage. Further, the decline in CD4+ percentage was due to a specific decrease in the suppressor-inducer subset of CD4 as defined using anti-CD45R. No change in the helper-effector subset of CD4 was noted. The percentage of CD8+ lymphocytes did not change significantly at any time postburn nor did subsets of CD8 as defined using anti-CD11. Numerical changes in lymphocyte subsets were dominated by a general lymphopenia occurring on Day 4 following injury. However, suppressor-inducer (CD4+/CD45R+) T cells also decreased significantly on postburn Day 1. These results further elucidate phenotypic changes in immunoregulatory subsets following major injury and suggest a possible basis for depressed autologous mixed lymphocyte responsiveness of burn patient T cells, one of the functional immunologic defects associated with severe injury.     

Moroccan lymphocyte subsets reference ranges: age,gender, ethnicity,and socio-economic factors dependent differences

ABSTRACT

Lymphocyte subsets reference ranges are helpful for a precise diagnosis and therapy of various diseases. We attempted in the current study to establish Moroccan lymphocyte reference range and reveal age, gender, ethnicity, income, and instructional levels dependent differences. Lymphocyte subsets percentage and absolute count were determined by 4-color flow cytometry in a population study of 145 adults Moroccan healthy volunteers. Analysis showed significant age-dependent changes. Age was associated with a decrease of naïve CD4+ and CD8+ T cells and an increase of memory CD4+ or CD8+ T cells. Activated CD4+ CD38+ and CD8+ CD38+ T cells, Treg as well as NK cell showed age-dependent alterations. In contrast, B cells remained unchanged. A higher percentage of CD3+ and CD4+ T cells was observed in females while CD8+, B and NK cells count were higher in men. Ethnicity, instructional levels, and personal income seem to not influence lymphocyte subsets reference values. This study provides reference ranges for lymphocyte subsets of healthy Moroccan adults. These results can be used for other North African (Maghrebian) countries considering their geographic, ethnic, economic, and cultural similarities.     

广州市健康儿童和成人淋巴细胞亚群数量的对比分析

目的比较儿童和成人淋巴细胞亚群百分率和绝对数量的异同,更好地为本地区临床诊断和治疗提供参考。方法采用双色流式细胞术分析健康儿童和成人外周血T淋巴细胞亚群(CD3+细胞、CD3+CD4+细胞、CD3+CD8+细胞)、B淋巴细胞(CD19+)、CD3-CD56+NK细胞(自然杀伤细胞)的数量。结果儿童组总淋巴细胞(百分率和绝对数)、CD19+淋巴细胞(百分率和绝对数)、CD3+淋巴细胞百分率、CD3+CD4+淋巴细胞百分率、CD3+CD8+淋巴细胞绝对数、CD3-CD56+细胞绝对数、CD4+/CD8+细胞比值与成人组比较差异均有统计学意义;CD3+CD8+淋巴细胞百分率、CD3-CD56+细胞百分率、CD3+淋巴细胞绝对数和CD3+CD4+淋巴细胞绝对数与成人组比较差异无统计学意义。儿童组CD3+CD8+淋巴细胞、CD19+淋巴细胞、CD3-CD56+细胞的百分率和绝对数均高于成人组,CD3+淋巴细胞(百分率和绝对值)、CD3+CD4+淋巴细胞(百分率和绝对值)和CD4+/CD8+细胞比值低于成人组。相同与不同性别组内均有多个指标存在显著差异。结论淋巴细胞亚群的分布受年龄、性别因素的影响,淋巴细胞亚群绝对数与百分率随年龄的变化不总是保持一致的。用于血液性和免疫性疾病诊断时,采用淋巴细胞亚群绝对数作为参考指标优于百分比率。     

重型乙型肝炎前期患者外周血免疫活性细胞的特点

目的 分析重型乙型肝炎前期(以下简称重肝前期)患者外周血免疫活性细胞的特点.方法 选取重肝前期患者48例,慢性乙型肝炎患者35例,健康志愿者20例,采用流式细胞仪检测外周血淋巴细胞CD3+、CD3+/CD4+、CD3 +/CD8+、CD4 +/CD25 +/CD45+等亚群表达百分比,计算各淋巴细胞亚群绝对值,并进行统计学分析.结果 与慢性乙肝组及健康对照组相比,重肝前期组CD3+T细胞、CD8+T细胞、CD4+ CD25+调节性T细胞(Tregs)的绝对值均明显下降(P＜0.01或P＜0.05);重肝前期组CD4+T细胞绝对值与慢性乙肝组差异无统计学意义(P＞0.05),但CD4+T细胞百分率明显高于慢性乙肝组(P＜0.05).此外,CD4 +/CD8+比值各组间存在显著差异,重肝前期组显著高于慢性乙肝组和健康对照组(P＜0.01或P＜0.05).结论 重型乙型肝炎前期存在一定程度的细胞免疫功能紊乱,其特征为CD4+T细胞占优势,CD8+T细胞和CD4+ CD25+ Tregs绝对值下降.     

Longitudinal follow-up of lymphocyte subsets during the first year of life

A longitudinal study of lymphocyte subsets during infancy was evaluated by using the flow cytometric immunophenotyping method. Two hundred and thirteen blood samples were obtained from 92 healthy, full-term infants of the following ages: 1-7 days old (n = 43), 3 months old (n = 55), 6 months old (n = 57) and 11 months old (n = 58). The absolute numbers of CD3+ and CD3+/CD4+ T lymphocytes increased from birth to 3 months of age, and remained stable thereafter. The absolute number of CD3+/CD8+ T lymphocytes increased from birth to 11 months of age. The absolute number of CD19+ B lymphocytes and NK cells increased rapidly (3 months) after birth and continued to increase throughout the study period. However, the changes in the relative counts of lymphocyte subsets did not always correspond with the changes in their absolute numbers. These results demonstrate the age-related changes in lymphocyte subpopulations and provide reference ranges for lymphocyte subsets during infancy.     

Normal values of peripheral lymphocyte populations and T cell subsets at a fixed time of day: a flow cytometric analysis with monoclonal antibodies in 210 healthy adults.

Using monoclonal antibodies and flow cytometry percentages and absolute number of lymphocyte populations and T cell subsets were enumerated in peripheral blood collected between 8:00 and 10:00 a.m. from 210 healthy adults. Although absolute numbers of total lymphocytes did not differ depending on age and sex, the numbers of T (Leu-4+) and natural killer (Leu-7+) cells as well as their percentages showed negative and positive correlations, respectively, with age. In Leu-7+ cells, the percentage was male-dominant irrespective of age, and the absolute number was male-dominant only in older subjects. Absolute numbers of suppressor/cytotoxic T (Leu-2a+) and helper/inducer T (Leu-3a+) cells and percentages of Leu-2a+ cells were negatively correlated with age in females. In males, only the percentage of Leu-2a+ cells was age-dependent. Leu-3a/Leu-2a was positively correlated with age in females, and was female-dominant depending on age. These results indicate that peripheral lymphocyte populations and T cell subsets vary remarkably in healthy adults even at a fixed time of day.     

Changes of Lymphocyte Subsets in Normal Pregnant and Postpartum Women: Postpartum Increase of NK/K (Leu 7) Cells

ABSTRACT: Changes in lymphocyte subsets in whole blood of normal pregnant and postpartum women were examined by flow cytometry with an automated leukocyte differential system. From the first trimester and throughout pregnancy, the absolute counts of T(CD3) and B(CD20) and T-cell subsets (CD4, CD8) decreased with a decrease in the absolute lymphocyte count, although the proportions of these cells remained unchanged except for a decrease in the percentage of T helper-inducer (CD4) cells in the first trimester. On the contrary, the percentage of NK/K (Leu 7) cells, but not of NK/K (CD16) cells, increased in the first trimester and then both gradually decreased in the second and third trimesters. In the postpartum period, the percentages and absolute counts of T(CD3) and NK/K (Leu 7) cells, but not of other cells, increased transiently. These changes of lymphocyte subsets may indicate suppression of immunological activity during pregnancy and its “increase” in the postpartum period.     

Lymphocyte subset analysis to predict progression to AIDS in a cohort of homosexual men in San Francisco

A group of 10 leukocyte and lymphocyte subsets were measured by simultaneous dual immunofluorescence and flow cytometry in a group of homosexual men from the San Francisco General cohort. Absolute numbers and percentages of lymphocytes were determined in 30 individuals who progressed to AIDS and 29 who did not over a 44-month period at annual intervals. At entry into the study, all subjects were asymptomatic, HIV seropositive, and had multiple changes in lymphocyte subsets compared to HIV-negative controls. In progressors, large changes occurred from the first visit to the last visit before progression in both absolute numbers and percentages of CD4 cells. The percentage of HLA-DR-bearing CD8 cells also increased. We utilized a proportional hazards model to assign a predictive value for progression to AIDS to lymphocyte subsets in both univariate and multivariate tests. Increased DR-positive CD8 cells, decreased CD4 cells, and increased CD8-positive, Leu 7-positive cells independently predicted progression to AIDS at P less than 0.006 (relative hazard 5.8-4.0). In a multivariate model, the most useful tests were either increased numbers or percentages of DR-positive CD8 cells. These data suggest parsimonious approaches to following HIV-positive individuals and further support the possibility of autoreactive T cells in the pathogenesis of HIV-associated diseases.     

T lymphocyte subsets and NK cell cytotoxicity in chronic hemodialysis patients. The effect of recombinant human erythropoietin (rHu-EPO) treatment.

We investigated subpopulations of T lymphocytes, NK cell number and cytotoxic activity in 14 chronic uremic patients on regular hemodialysis treatment. We observed a significantly decreased absolute lymphocyte number and percentage of CD3 cells. Relative numbers of CD16 cells were significantly elevated, but NK cell cytotoxic activity was within a normal range. Nine patients with chronic renal anemia on maintenance hemodialysis were enrolled in rHu-EPO treatment trial. The treatment was continued till the hematocrit level reached 30%. Each of the patients had corrected anemia and well-being. After 12 weeks of the treatment we observed in these patients decreases in CD3, CD4, CD8 and CD16 cell numbers and elevation of CD4/CD8 ratio. Cytotoxic activity of NK cells did not change significantly. Presented results indicate that chronic hemodialysis patients have significantly diminished lymphocyte number. rHu EPO treatment affects the T lymphocyte subsets inducing a deep decrease of CD8 and CD16 cell percentage leading to normalisation of the CD4/CD8 ratio.     

Abnormal distribution of distinct lymphocyte subsets in children with Wiskott-Aldrich syndrome

Wiskott-Aldrich syndrome (WAS) is a severe and rare primary immunodeficiency. Several studies show that WAS protein (WASp) plays a key role in the function of certain lymphocyte subsets. So far, no study has described distinct immunophenotypic abnormalities associated with WAS; thus the prognostic significance of any such abnormalities is unclear. This study examined many differences in the percentage/absolute numbers of distinct lymphocyte subsets in 20 WAS patients and 20 age/sex-matched healthy controls, and analyzed the association between these abnormalities and clinical disease scores. The results showed that the numbers of CD4⁺ T cells, B cells, and CD8⁺ naïve T cells were significantly lower in WAS patients; furthermore, the numbers in WASp-negative patients were lower than those in WASp-positive patients. WAS patients showed a selective reduction in expression of CD19 by naïve and transitional B cells. There was a negative association between the number of B cells and the WAS clinical scores. Also, CD8⁺ naïve T cell numbers in patients with a score of 3–5A were lower than those in patients with a score of 2. The absence of WASp leads to a reduction in the population of specific lymphocyte subsets; therefore, these findings may help future management of patients with WAS.     

Changes in lymphocyte subpopulations and CD3+/DR+ expression in sepsis

Objective   To detect lymphocyte subpopulations and CD3+/DR + expression in sepsis.
Methods   In a prospective clinical study we evaluated subpopulations of lymphocytes and percentage of CD3⁺/HLA-DR⁺ lymphocytes using two-color flow cytometry in 40 patients with sepsis and compared them with 34 healthy adults.
Results   Septic patients, when compared with healthy controls, have significantly lower percentage and absolute numbers of total T lymphocytes and CD4 T lymphocytes ( P  < 0.01). Absolute numbers of CD8 T lymphocytes, NK cells, CD3+/DR + lymphocytes and CD4/CD8 ratio were also decreased ( P  < 0.01). The percentage of B lymphocytes was increased ( P  < 0.01).
Conclusion   Our results are in agreement with previous findings in patients with sepsis after major surgery or trauma. The decreases in the percentage and absolute numbers of circulating lymphocyte subsets in non-surgical sepsis could represent a general reaction to stress. Increased percentage of B lymphocytes is most probably related to the bacterial etiology of the disease.     

Immunophenotyping of blood lymphocytes at birth, during childhood, and during adulthood in HIV-1-uninfected Ethiopians

To obtain more insight into blood lymphocyte subpopulations of Ethiopians, we studied the immunologic profile of children and neonates and compared these data with those obtained from adults. Peripheral blood mononuclear cells (PBMCs) and cord blood mononuclear cells (CBMCs) were collected from 137 HIV-1-uninfected subjects aged 0 (cord blood) up to 40 years. Lymphocyte subsets (T, B, and NK cells, CD4+ and CD8+ T cells) were determined and T cell activation (CD38 and HLA-DR) and differentiation (CD45RO and CD27) markers were measured on CD4+ and CD8+ T cells. The absolute number and percentage values of most lymphocyte subpopulations differed substantially with age. Neonates and children were found to have significantly higher CD4+ T cell counts compared to adults. The median absolute CD4 count at birth was comparable to those reported for Caucasians. At birth 97% of the CD4+ T cells were na?ve and this proportion significantly declined to 14.2% during adulthood. In addition, activation of both CD4+ and CD8+ T cells, as determined by the double expression of HLA-DR and CD38, was observed in children under the age of 16 and adults, but not in neonates. A more differentiated phenotype (CD27-) was observed in adults compared to children for both CD4+ and CD8+ T cells. The immune alterations including the remarkably low CD4 count with highly depleted na?ve phenotype and a persistently activated immune system seen in adult Ethiopians are not apparent at birth, but rather develop over time.     

Determination of laboratory reference ranges for lymphocyte immunophenotype subsets in healthy adult Japanese using flow cytometry

This study represents a comprehensive evaluation of normative values for lymphocyte immunophenotype subsets using flow cytometry techniques in a Japanese population. Lymphocyte reference ranges were determined for percentage and absolute count of T, B, and NK cells in healthy adult Japanese using an extensive two-color immunophenotyping panel and consistently applied quality control methodology. Reference values were also determined for activation markers on CD3+ lymphocytes CD3+/CD25+, CD3+/CD38+ and CD3+/HLA-DR+. Differences in age and gender were observed for specific lymphocyte subsets. Comparison of the Japanese study with a Thai multi-center study that used similar methodology also demonstrated ethnic differences in lymphocyte reference ranges. The results in this study strongly suggest that reference values derived from studies in one population may not be applied to another population even when similar protocols for reagents, instruments and procedures are used although such studies do appear useful for epidemiological comparisons.     

Are the reference values of B cell subpopulations used in adults for classification of common variable immunodeficiencies appropriate for children?

Detailed phenotypic characterization of B cell subpopulations is of utmost importance for the diagnosis and management of humoral immunodeficiencies, as they are used for classification of common variable immunodeficiencies. Since age-specific reference values remain scarce in the literature, we analysed by flow cytometry the proportions and absolute values of total, memory, switched memory and CD21(-/low) B cells in blood samples from 168 healthy children (1 day to 18 years) with special attention to the different subpopulations of CD21(low) B cells. The percentages of total memory B cells and their subsets significantly increased up to 5-10 years. In contrast, the percentages of immature CD21(-) B cells and of immature transitional CD21(low)CD38(hi) B cells decreased progressively with age, whereas the percentage of CD21(low) CD38(low) B cells remained stable during childhood. Our data stress the importance of age-specific reference values for the correct interpretation of B cell subsets in children as a diagnostic tool in immunodeficiencies.     

Low Number of Peripheral Blood B Lymphocytes in Patients with Pulmonary Tuberculosis

The cellular immune response plays a critical role in the containment of persistent Mycobacterium tuberculosis infection; however, the immunological mechanisms that lead to its control are not completely identified. The goal of this study was to evaluate B (CD19+) and T (CD3+) peripheral blood lymphocyte profiles and T-cell subsets (CD4+ and CD8+) in patients with pulmonary tuberculosis (TB). Percentages (p = 0.02) and absolute numbers (p = 0.005) of B cells were significantly lower in patients with pulmonary TB than in healthy donors. In contrast, percentages (p = 0.12) and absolute numbers (p = 0.14) of T cells were similar in TB patients and healthy donors. No significant differences in percentages of CD4+ (p = 0.19) or CD8+ (p = 0.85) T cells between patients and healthy donors were observed. In summary, patients with pulmonary tuberculosis had a lower number of peripheral blood B lymphocytes than healthy controls.     

Lymphocyte subsets' reference ranges in an age- and gender-balanced population of 100 healthy adults--a monocentric German study

Region-specific reference ranges for adult peripheral blood lymphocyte subsets have been established in few countries around the world, but most studies were restricted to younger adults for monitoring of HIV patients. The aim of this investigation was to establish age- and gender-specific reference ranges for healthy adults. Lymphocyte subsets were examined in 100 healthy volunteers (50 males, 50 females) aged 19-85 years by two-color flow cytometric analysis with a FACSCalibur. A statistically significant decline in the mean numbers of CD3+/CD8+ T cells and CD19+ B cells was observed beyond an age of 50 years, whereas the mean counts of NK cells and CD4+/CD8+ ratio significantly increased beyond the age of 50. Females < or = 50 years had significantly higher mean CD4+ T cell counts and lower NK cell counts than males < or = 50 years. Based on these results, we established reference values for three subgroups: males < or = 50 years, females < or = 50 years, and males/females > 50 years.