Lymphocyte subpopulations, cytokines and trace elements in asymptomatic atopic women exposed to an urban environment

This study evaluates the immune response to exposure to an urban environment from 30 non-atopic and 30 non-symptomatic women with history of respiratory and/or cutaneous allergies. Blood lymphocyte subsets and serum interleukin (IL) 4 and interferon gamma (INF-gamma) of the two groups were similar, while serum IgE and "in vitro" production of IL-4 and INF-gamma by mononuclear blood cells of the atopic women were higher spontaneously or in the presence of PHA, respectively. Blood lead of the nonatopic women (mean 55 microg/l) was positively correlated with CD4+-CD45RO-, CD3+-CD8+ and CD3--HLA-DR+ lymphocyte subsets, while urinary trans-trans muconic acid (a metabolite of benzene) of both groups of women (mean about 50 microg/l) was significantly correlated with NK CD16+CD56+ lymphocytes. Urine chromium of the non-atopic subjects was significantly correlated with activated T, B and NK HLA-DR+ cells. Urine nickel of both groups of women was correlated with CD4+-CD45RO+ "memory" lymphocytes and their ratio with CD4+-CD45RO- "virgin" lymphocytes suggesting that the metal enhances maturation of "virgin" into "memory" lymphocytes. On the whole, this study demonstrates that exposure to low levels of toxic agents, produced by vehicular traffic in an urban environment, exerts effects on immune functions of women.     

Circulating leucocyte and lymphocyte subpopulations before and after intensive endurance exercise to exhaustion.

Seventeen healthy cyclists [age 20.8 (SD 4.8) years; body mass 68.3 (SD 7.7) kg; body fat, 11.4 (SD 2.6) %; height, 179.1 (SD 5.9) cm; VO2max, 60.9 (SD 7.4) ml.kg-1.min-1] conducted intensive endurance exercise to exhaustion (stress test, ST) on a cycle ergometer at 110% of their individual anaerobic threshold [Than,individual; exercise intensity, 3.97 (SD 0.6) W.kg-1; duration, 23.9 (SD 8.3) min; maximal lactate concentration, 7.39 (SD 2.59) mmol.l-1]. The distribution of leucocyte subpopulations was measured flow cytometrically: before, immediately after (0), 5 (+5), 30 (+30) and 60 (+60) min after ST. The lymphocytes (0 min) and granulocytes (+60 min) were mainly responsible for the increase of leucocytes. Lymphocytes were significantly lower at +30 and +60 min than before. CD3-CD16/CD56+ (+480%) and CD8(+)-lymphocytes (+211%) increased at 0 min more than the other lymphocyte subpopulations (CD(3+)-cells, +100%; CD(4+)-cells, +56%; CD(19+)-cells, +64%). CD3-CD16/CD(56+)- and CD(8+)-cells also were mainly responsible for the decreased values of lymphocytes at +30 min and +60 min compared to before. At 0 min naive CD(8+)-cells (CD45RA+, CD45RO-) increased more than memory CD(8+)-cells (CD45RA-, CD45RO+). Changes of naive and memory CD(4+)-cells did not differ. All lymphocyte subpopulations, in particular CD(8+)- and CD3-CD16/CD(56+)-cells, decreased rapidly between 0 min and 5 min. We conclude that an intensive endurance exercise to exhaustion causes a mobilisation of lymphocytes, especially of natural killer cells (CD3-CD16/CD56+) and naive, unprimed CD(8+)-cells (CD45RA+, CD45RO-) which may be transported to injured muscles.(ABSTRACT TRUNCATED AT 250 WORDS)     

The relationship of CD16 (Leu-11) and Leu-19 (NKH-1) antigen expression on human peripheral blood NK cells and cytotoxic T lymphocytes

We examined the antigenic and functional characteristics of human peripheral blood lymphocytes that differentially express the CD16 (Leu-11) and Leu-19 (NKH-1) antigens. Leu-19 is a approximately 220,000 daltons protein expressed on approximately 15% of freshly isolated peripheral blood lymphocytes. Within the Leu-19+ subset, three distinct populations were identified: CD3-,CD16+,Leu-19+ cells; CD3+,CD16-,Leu-19+ cells; and CD3-,CD16-,Leu-19bright+ cells. Both the CD3+,CD16-,Leu-19+ and CD3-,CD16+,Leu-19+ populations mediated non-major histocompatibility complex (MHC)-restricted cytotoxicity against the NK-sensitive tumor cell K562 and were large granular lymphocytes. CD3-,CD16+,Leu-19+ NK cells were the most abundant (comprising approximately 10% of peripheral blood lymphocytes) and the most efficient cytotoxic effectors. The finding that CD3+,Leu 19+ lymphocytes mediated cytotoxicity against K562 unequivocally demonstrates that a unique subset of non-MHC-restricted cytotoxic CD3+ T lymphocytes are present in the peripheral blood of unprimed, normal individuals. However, CD3+,CD16-,Leu-19+ cells comprised less than 5% of peripheral blood lymphocytes, and the cytotoxic activity of this subset was significantly less than CD3-,CD16+,Leu-19+ NK cells. Most CD3+,Leu-19+ T cells co-expressed the CD2, CD8, and CD5 differentiation antigens. The antigenic and functional phenotype of peripheral blood CD3+,Leu-19+ cytotoxic T lymphocytes corresponds to the interleukin 2-dependent CD3+ cell lines that mediate non-MHC-restricted cytotoxicity against NK-sensitive tumor cell targets. A small population of Leu-19bright+ lymphocytes lacking both CD3 and CD16 was also observed. This population (comprising less than 2% of peripheral blood lymphocytes) contained both large agranular lymphocytes and large granular lymphocytes. CD3-,CD16-,Leu-19bright+ lymphocytes also mediate non-MHC-restricted cytotoxicity. The relationship of these CD3-CD16-,Leu-19bright+ lymphocytes to CD3+ T cells or CD16+ NK cells is unknown.     

Effects of resistance training on selected indexes of immune function in elderly women.

Women aged 67-84 yr were randomly assigned to either resistance exercise (RE, n = 15) or control group (C, n = 14). RE group completed 10 wk of resistance training, whereas C group maintained normal activity. Blood samples were obtained from the RE group (at the same time points as for resting C) at rest, immediately after resistance exercise, and 2 h after exercise before (week 0) and after (week 10) training. Mononuclear cell (CD3+, CD3+CD4+, CD3+CD8+, CD19+, and CD3-CD16+CD56+) number, lymphocyte proliferative (LP) response to mitogen, natural cell-mediated cytotoxicity (NCMC), and serum cortisol levels were determined. Strength increased significantly in RE subjects (%change 8-repetition maximum = 148%). No significant group, exercise time, or training effects were found for CD3+, CD3+CD4+, or CD3+CD8+ cells, but there was a significant exercise time effect for CD3-CD16+CD56+ cells. LP response was not different between groups, across exercise time, or after training. NCMC was increased immediately after exercise for RE subjects at week 0 and for RE and C groups at week 10. The week 0 and week 10 NCMC values were above baseline for both RE and C groups 2 h after exercise. In conclusion, acute resistance exercise did not result in postexercise suppression of NCMC or LP, and 10 wk of resistance training did not influence resting immune measures in women aged 67-84 yr.     

A comprehensive study on neurobehavior, neurotransmitters and lymphocyte subsets alteration of Chinese manganese welding workers

The neurotoxicity of manganese has been demonstrated by many researches. But few reports have been found on its immunotoxicity in manganese-exposed workers. Here we selected welding workers (aged 34 years) as Mn-exposed subjects. They have been exposed to manganese for 16 years. The control group was from a flour plant. The average concentrations of Mn, Cd, Fe and Ni in work place were 138.40 +/- 11.60 microg/m3, 581.40 +/- 45.32 microg/m3, 3.84 +/- 0.53 microg/m3 and 12.64 +/- 2.80 ng/m3, respectively. Blood Mn (4.84 mug/dl) of welding workers was higher than that of the control group (1.92 microg/dl). Neurobehavioral core test battery (NCTB) recommended by WHO was conducted on the subjects and found that the scores of negative emotions, such as confusion-bewilderment, depression-dejection, fatigue-inertia, and tension-anxiety, were higher in welding workers. Visual simple reaction time and the fast simple reaction time were shorter than that of the control group. The numbers of digital span, forward digital span, backward digital span and digital symbol decreased in welding workers compared with control group. Monoamine neurotransmitters and their metabolism substances in urine were tested by HPLC-ultraviolet. NE, E, MHPG, HVA, DA, DOPAC and 5-HT in the urine of Mn-exposed group had no significant changes while 5-HIAA in Mn-exposed group had significantly decreased compared with that of the control group. Lymphocyte subsets of the subjects were determined by Flow Cytometer. CD3+ T cell, CD4+CD8- T cell, CD4-CD8+ T cell, CD4+CD45RO- "virgin" lymphocytes, CD4+CD45RO+ "memory" lymphocytes, and CD3-CD19+ B cell had no significant changes compared with the control group. The results showed that long-term exposure to manganese in welding might have adverse effects on mood state, neurobehavior, and peripheral neurotransmitters. However, they had no effects on lymphocyte subsets parameters.     

Differential mobilization of leucocyte and lymphocyte subpopulations into the circulation during endurance exercise.

A total of 14 healthy subjects [means (SD): 27.6 (3.8) years; body mass 77.8 (6.6) kg; height 183 (6) cm] performed endurance exercise to exhaustion at 100% of the individual anaerobic threshold (Th(an)) on a cycle ergometer (mean workload 207 (55) W; lactate concentrations 3.4 (1.2) mmol.l-1; duration 83.8 (22.2) min, including 5 min at 50% of individual Th(an)). Leucocyte subpopulations were measured by flow cytometry and catecholamines by radioimmunological methods. Blood samples were taken before and several times during exercise. Values were corrected for plasma volume changes and analysed using ANOVA for repeated measures. During the first 10 min of exercise, of all cell subpopulations the natural killer cells (CD3-CD16/CD56+) increased the most (229%). Also CD3+CD16/CD56+ (84%), CD8+CD45RO- (69%) cells, eosinophils (36%) and monocytes (62%) increased rapidly during that time. CD3+, CD3+HLA-DR+, CD4+CD45RO+, CD4+CD45RO-, CD8+CD45RO+ and CD19+ cells either did not increase or increased only slightly during exercise. Adrenaline and noradrenaline increased nearly linearly by 36% and 77% respectively at 10 min exercise. The increase of natural killer cells and heart rates between rest and 10 min of exercise correlated significantly (r = 0.576, P = 0.031). We conclude that natural killer cells, cytotoxic, non-MHC-restricted T-cells, monocytes and eosinophils are mobilized rapidly during the first minutes of endurance exercise. Both catecholamines and increased blood flow are likely to contribute this effect.     

Immune status and proinflammatury cytokines in pregnant women with acute cytomegalovirus infection

A total of 64 pregnant women aged 16-40 years with acute cytomegalovirus (CMV) infection were examined. CMV infection was diagnosed by the detection of anti- CMV IgM (by the enzyme immunoassay) or DNA by PCR in the blood. The immunophenotype of lymphocytes CD3+, CD4+, CD8+, CD16+, CD19+, IgG, IgM, IgA and the spontaneous production of TNFalpha and IL-8 in the blood were studied. In pregnant women with CMV infection the presence of the immunosuppression of cell-mediated immunity reactions, the hyperproduction of Fc-receptors of natural killers, B lymphocytes (CD19+), the hyperglobulinemia of serum IgM and IgG, as well as the increased synthesis of proinflammatory cytokines--TNFalpha and IL-8--were established.     

Identification and analysis of a novel human surface CD5- B lymphocyte subset producing natural antibodies.

The production of "natural" autoantibodies or antibodies, i.e., Ig that bind a variety of self- and/or exogenous Ag and arise independently of known immunization, is though to be a feature of CD5+ B lymphocytes. To determine whether other lymphocyte subsets exist that might be committed to the production of natural antibodies, human peripheral blood B cells were sorted on the basis of surface CD5 expression and differential expression of surface CD45RA (CD5+CD45RAintermediate(int), CD5-CD45RAlow(lo), and CD5-CD45RAhigh(hi)), and analyzed for the type of Ig produced after EBV infection and culture. Like their CD5+ counterparts, most CD5-CD45RAlo B lymphocytes were precursors of cells producing IgM, a major proportion of which displayed the Ag-binding features of natural antibodies. In contrast, CD5-CD45RAhi B cells comprised a high frequency of IgG-producing cell precursors, possibly including memory B lymphocytes. Six of seven IgM mAb generated from sorted CD5-CD45RAlo B cells and three of four IgM mAb from sorted CD5+ B cells were polyreactive, binding with different affinities (Kd, 10(-5) to 10(-8) M) to two or more Ag; the remaining mAb from CD5-CD45RAlo and the mAb from CD5+ B cells each bound to a single Ag (Kd, 10(-7) to 10(-8) M), beta-galactosidase and ssDNA, respectively. CD5-CD45RAlo B cells account for 4.1 +/- 1.2% (mean +/- SD in 11 healthy subjects; CD5+ B cells, 23.3 +/- 6.9%) of total B lymphocytes and display the features of quiescent cells. In a given individual, the number of CD5-CD45RAlo B cells remains constant over time. CD5-CD45RAlo and CD5+ B cells bear surface CD11b and CD14, at densities and/or frequencies apparently higher than those of CD5-CD45RAhi B lymphocytes. Despite their surface CD5- phenotype, CD45RAlo B cells express CD5+ mRNA at levels comparable with those of CD5+ B lymphocytes, whereas CD5-CD45RAhi B cells express only trace amounts of CD5 mRNA. The commitment to natural antibody production and the degree of CD5 mRNA expression suggest that the newly defined CD5-CD45RAlo B cell subset is related to CD5+ B lymphocytes, and may constitute the human homologue of the mouse Ly-1-"sister" B cell population.     

Tumor-selective cytolysis is executed exclusively by CD45R+ CTL whereas allo-specific cytotoxicity can be executed also by CD45R- CTL

Naive and memory CD4+ T helper cells can be distinguished on the basis of expression of the CD45R molecule. Whether this dichotomy applies also to CD8+ T cells has not yet been established. In the present investigation the cytolytic activity of peritoneal CD8+CD45R+ and CD8+CD45R- T cells from tumor- and allo-immunized rats has been studied. More than 90% of the CD8+ peripheral blood T lymphocytes expressed the CD45R molecule, whereas in the peritoneal cavity about 60% of the CD8+ T cells displayed the CD45R+ phenotype. Analysis of cytotoxicity of sorted peritoneal cells of W439 tumor-immunized donors demonstrated selective cytolytic activity of the CD5+CD4-CD8+CD45R+ subpopulation to W439 lymphoma target cells but no effect of CD5+CD4-CD8+CD45R- lymphocytes. None of these lymphocyte populations exhibited cytolytic activity to the NK-sensitive cell line YAC-1, whereas the CD5-CD45R+ population showed strong cytotoxicity to YAC-1 cells. In allo-immunized rats both CD5+CD4- CD8+CD45R+ and CD5+CD4-CD8+CD45R- peritoneal cells exhibited strong allo-specific cytolytic activity, but no activity to YAC-1 cells. Both CD5+CD4-CD8+CD45R+ and CD5+CD4-CD8+CD45R- cells from tumor-immunized rats proliferated in response to Con A and rIL-2. This is the first study demonstrating that tumor-selective cytolytic CD8+ T cells express the CD45R molecule and that allo-specific cytolytic CD8+ T cells are found in both the CD45R+ and CD45R- populations.     

Mobilization of circulating leucocyte and lymphocyte subpopulations during and after short, anaerobic exercise

A group of 11 healthy athletes [age, 27.4 (SD 6.7) years; body mass, 75.3 (SD 9.2) kg; height, 182 (SD 8) cm; maximal oxygen uptake, 58.0 (SD 9.9) ml.kg-1.min-1] conducted maximal exercise of 60-s duration on a cycle ergometer [mean exercise intensity, 520 (SD 72) W; maximal lactate concentration, 12.26 (SD 1.35) mmol.l-1]. Adrenaline and noradrenaline, and leucocyte subpopulations were measured flow cytometrically at rest, after 5-min warming up at 50% of each individual's anaerobic threshold (followed by 5-min rest), immediately after (0 min), 15 min, 30 min, and 1, 2, 4 and 24 h after exercise. Granulocytes showed two increases, the first at 15 min and, after return to pre-exercise values, the second more than 2 h after exercise. Eosinophils also increased at 15 min but decreased below pre-exercise values 2 h after exercise. Total lymphocytes and monocytes had their maximal increases at 0 min. Out of all lymphocyte subpopulations CD3-CD16/CD56(+)- and CD8+CD45RO--cells increased most and had their maximal cell counts at 0 min. The CD3(+)-, CD4+CD45RO(+)-, CD8+CD45RO(+)-, and CD19(+)- increased at 0 min, but had their maximum at 15 min. During the hours after exercise CD3-CD16/CD56(+)-, CD3+CD16/CD56(+)-, CD8+CD45RO(+)- and CD8+CD45RO--cells were responsible for the lymphocytopenia. The CD3(+)- and CD3-CD16/CD56(+)-cells were lower 24 h after exercise than before exercise. Adrenaline and noradrenaline increased during exercise. In conclusion, short anaerobic exercise led to a sequential mobilization of leucocyte subpopulations. The rapid increase of natural killer cells and monocytes may have been due to increased blood flow and catecholamine concentrations.(ABSTRACT TRUNCATED AT 250 WORDS)     

In situ localization of CD45 isoforms in the human thymus indicates a medullary location for the thymic generative lineage.

Previous work has suggested that the generative lineage within the human thymus can be defined by the selective expression of CD45 isoforms and is CD45RO- and predominantly CD45RA+. In order to physically localize these cells we have stained frozen sections of human thymus with antibodies to CD45RO (p180), and CD45RA (p205/P220), as well as with CD1 and HLA class I to define cortical and medullary areas, respectively. In the cortex, 70 to 90% of thymocytes were CD45RO+, whereas only 0.5% expressed CD45RA. Medullary cells were 30% CD45RO+, 29% CD45RA+; approximately 40% did not express detectable levels of either isoform but did express CD45 common determinants. To assess the degree of proliferation of cells expressing CD45 isoforms, we stained adjacent sections, or used double staining, with Ki67, an antibody that detects a nuclear Ag on proliferating cells. We found that CD45RA+ thymocytes are predominantly a resting medullary population with a small component in cell cycle, consistent with our analysis of human thymocytes by immunofluorescence, and with data in murine systems defining the generative lineage. To confirm that the CD1- or low, CD45RO-CD45RA+ thymocytes defined by immunofluorescence analysis were likely to have a medullary location, we analyzed the CD4/CD8 subset distribution of CD1-cells. From 80 to 90% of CD1-thymocytes are CD4+ or CD8+ single positives or CD-8- double negatives. CD1-thymocytes also include 12 to 14% CD4+8+ cells with a probable medullary location. A similar analysis of lymphocytes expressing a high density of HLA class I, which have a medullary location, confirmed the existence of CD4+8+ thymocytes in the medulla. Purified CD3-4-8- cells, previously shown to be CD1-CD45RA+, were also shown to bear a high density of HLA class I, indicating a medullary location. Correlative localization of a panel of Ag thus supports the argument for a medullary location of the thymic generative lineage.     

香菇多糖注射液对直肠癌根治术后患者纤维蛋白水平及免疫功能的影响

目的:研究香菇多糖注射液对直肠癌根治术后患者纤维蛋白水平及免疫功能的影响。方法:将2010年5月-2014年10月期间在我院接受直肠癌根治术的患者随机分为两组,观察组术后接受香菇多糖联合FOLFOX-4化疗,对照组术后接受FOLFOX-4化疗,观察两组患者化疗过程中的不良反应,测定化疗前后纤维蛋白水平及免疫功能指标。结果:观察组患者在化疗过程中消化道反应、肝功能损伤、骨髓抑制、周围神经毒性、口腔黏膜损伤、脱发的发生率均显著低于对照组(P0.05);化疗后,对照组患者外周血中CD3~+CD4~+CD8~-细胞、CD3~+CD4~-CD8~+细胞、CD19~+细胞、CD3~-CD56~+细胞的含量以及血清Ig M、Ig A、Ig G、FIB的含量显著低于化疗前(P0.05),观察组患者外周血中CD3~+CD4~+CD8~-细胞、CD3~+CD4~-CD8~+细胞、CD19~+细胞、CD3~-CD56~+细胞的含量以及血清Ig M、Ig A、Ig G的含量与化疗前无显著性差异(P0.05),血清FIB含量显著低于化疗前(P0.05);观察组患者化疗后外周血中CD3~+CD4~+CD8~-细胞、CD3~+CD4~-CD8~+细胞、CD19~+细胞、CD3~-CD56~+细胞的含量以及血清Ig M、Ig A、Ig G、FIB的含量显著高于对照组(P0.05),血清FIB含量显著低于对照组(P0.05)。结论:香菇多糖注射液辅助治疗能够减轻直肠癌根治术后FOLFOX-4化疗的毒副作用、改善免疫功能。     

Stepwise differentiation of CD4 memory T cells defined by expression of CCR7 and CD27

To study the steps in the differentiation of human memory CD4 T cells, we characterized the functional and lineage relationships of three distinct memory CD4 subpopulations distinguished by their expression of the cysteine chemokine receptor CCR7 and the TNFR family member CD27. Using the combination of these phenotypic markers, three populations were defined: the CCR7+CD27+, the CCR7-CD27+, and the CCR7-CD27- population. In vitro stimulation led to a stepwise differentiation from naive to CCR7+CD27+ to CCR7-CD27+ to CCR7-CD27-. Telomere length in these subsets differed significantly (CCR7+CD27+ > CCR7-CD27+ > CCR7-CD27-), suggesting that these subsets constituted a differentiative pathway with progressive telomere shortening reflecting antecedent in vivo proliferation. The in vitro proliferative response of these populations declined, and their susceptibility to apoptosis increased progressively along this differentiation pathway. Cytokine secretion showed a differential functional capacity of these subsets. High production of IL-10 was only observed in CCR7+CD27+, whereas IFN-gamma was produced by CCR7-CD27+ and to a slightly lesser extent by CCR7-CD27- T cells. IL-4 secretion was predominantly conducted by CCR7-CD27- memory CD4 T cells. Thus, by using both CCR7 and CD27, distinct maturational stages of CD4 memory T cells with different functional activities were defined.     

Postthymic development of CD28-CD8+ T cell subset: age-associated expansion and shift from memory to naive phenotype

During human aging, one of the major changes in the T cell repertoire is a dramatic expansion of T cells with the atypical CD28-CD8+ phenotype. In this study, we show that this increase is a consequence not only of an expansion in the CD28-CD8+ population but also of a decrease in the number of CD28+CD8+ T cells. The decrease in circulating CD28+CD8+ T cells is dramatically accelerated after the age of 50 and is not accompanied by an equivalent reduction in the CD28+CD8+ subset. Our findings confirm that aging leads to an accumulation of CD45RO+ T cells within the CD28+CD8+ subset as previously observed. Surprisingly, we found an increase in CD45RA+ expression with age in the CD28-CD8+ subset. Immune-phenotyping for activation markers, measurement of telomere DNA content, and cytokine production analysis indicate that the large majority of CD28-CD8+ T cells are Ag-experienced, despite their CD45RA+ phenotype. Our study further demonstrates that the poor proliferative response displayed by CD28-CD8+ T cells is not a consequence of telomere shortening. Also, analysis of cytokine production at the single cell level revealed that the proportions of IFN-gamma +, IL-4+, and IL-10+ T cells are considerably higher among the CD28-CD8+ than the CD28+CD8+ subset. In summary, these data explain the presence of CD45RA+ T cells in the elderly, shed light on the phylogenetic origin of CD28-CD8+ T cells, and suggest a role for these cells in the immune senescence process.     

Selection of CD4+CD8+ thymocytes by complex formation with medulla-derived epithelial cells

The role of lymphostromal complexes in T-cell differentiation is far from elucidated, mainly because a clear association of a particular stromal cell type with a distinct thymocyte subset has never been identified. Using an in vitro system, detecting the adherence of thymocytes to a thymic medullary epithelial cell line (E-5), we showed that the phenotype of these thymocytes was that of cortical type: Thy-1hi, LFA-1+, PNAhi, CD4+CD8+, MEL-14-/lo, IL-2R-, CD3-/lo, and TcR V beta 8-/lo. They were enriched in cells in G2/M at the time of complex formation, showed a higher basal proliferation in culture, and did not respond to PHA, IL-2 and only marginally to Con A. These data show that complex formation with mouse thymic medullary epithelium selects for CD4+CD8+ thymocytes, as shown by the marked decrease in CD4+CD8-/CD4-CD8+ thymocytes, and the incapacity of CD4-CD8- thymocytes to adhere.     

The activation processes of the immune system during low intensity exercise without relaxation

The activation processes of T-, B- and NK-cells were studied during 8-week low intensity strength training without relaxation. After the long-term training, no changes occurred in the peripheral blood contents of the main subpopulations of immunocompetent cells (CD3+, CD4+, CD8+, CD19+ and CD16+/CD56+ lymphocytes) and serum levels of immunoglobulin A, M, and G. At the same time, training was accompanied by positive activation of the immunocompetent cells which was evident from increased percentage of CD3+, CD19+ and /CD56+ cells expressing CD69 after activation (PHA, PW and II.2, respectively). The final period of the training course was also associated with a decrease in the level of lymphocyte apoptosis and increase of proliferative responses of lymphocytes.     

Phenotype of peripheral blood leukocytes and survival of patients with metastatic colorectal cancer

Immune dysfunction is prevalent in metastatic cancer. Few patients with colorectal cancer metastases are cured, and among the strategies aimed at improving the therapeutic results in patients with metastatic colorectal cancer, immunotherapy is being increasingly investigated. We evaluated retrospectively the prognostic significance of peripheral blood leukocytes in 59 patients with metastatic colorectal cancer. The relative numbers of CD3+, CD3+CD4+, CD3+CD8+, NK (CD3-CD16+CD56+), CD3+DR+, CD3+CD25+, CD3+CD69+, CD19+, CD19+CD23+, CD8+CD28+, CD8-CD28+, CD8+CD57+, CD14+DR+ and CD14+CD16+ leukocytes were analyzed by two-color flow cytometry. A three-step approach was adopted to identify predictors of prognosis using regression analysis. Based on the results of univariate survival analysis, the absolute number of white blood cells, NK/CD3+CD69+ and NK/white cell count ratios were significant indicators of prognosis. In the multivariate regression analysis a model was obtained using a single parameter, the NK/CD3+CD69+ ratio, predicting the survival with 10-15% power of regression. The present results indicate that the NK/CD3+CD69+ ratio in peripheral blood may be an independent variable in a regression model predicting the overall survival of patients with colorectal cancer metastases to be tested in prospective studies.     

The abnormal apoptosis of T cell subsets and possible involvement of IL-10 in systemic lupus erythematosus

To study the apoptosis of lymphocyte subpopulations in systemic lupus erythematosus (SLE) patients and the possible role of IL-10 in this apoptosis involved in the pathogenesis of SLE, three color fluorescence and flow cytometry were used to investigate the early apoptosis of lymphocyte subsets from freshly separated or cultured peripheral blood mononuclear cells (PBMCs). ELISA was employed to detect the levels of IL-10 in serum and the levels of sFas and sFasL in cultured PBMC supernatants, and the results of sFas and sFasL were confirmed by real-time PCR of Fas and FasL mRNA. The results showed that in cells from SLE patients, the apoptosis of CD3+, CD4+, and CD8+ T cells was distinctly increased, and the percentage of CD4+ cells and the CD4/CD8 ratio was significantly decreased, as compared with normal controls. The apoptosis of T lymphocytes cultured with SLE serum was markedly higher than that of cells cultured with control's serum. Blockade of interleukin-10 (IL-10) activation by an anti-IL-10 antibody reduced the SLE serum induced apoptosis of CD4+ and CD8+ T cells. The levels of sFas and sFasL in the culture supernatant and Fas and FasL mRNA expressions in cultured cells were significantly higher in the SLE serum-cultured groups, but decreased evidently in the presence of the anti-IL-10 antibody. Above findings suggested that SLE cells showed abnormally high apoptosis of T lymphocytes, especially of the CD4+ subpopulation, resulting in a decreased CD4/CD8 ratio. The high percentage of apoptotic T cells in SLE patients may be related to the high levels of IL-10 in SLE serum, as IL-10 may induce the abnormally activated T cells to trigger apoptosis via the Fas-FasL pathway.     

IL-6/BSF-2 selectively stimulates the GO----S progression of CD8+ lymphocytes.

IL-6 preferentially promotes the DNA synthesis of human peripheral blood CD8+, rather than CD4+, lymphocytes in presence of PHA: this effect is observed in serum-free cultures of greater than 99% purified CD8+ lymphocytes. However, IL-6 is able to stimulate DNA synthesis of CD8+ lymphocytes triggered by a mitogenic anti-CD2 mAb, but not by anti-CD3 mAb: these results suggest that IL-6 selectively induces activation of CD8+ lymphocytes through the CD2 rather than the CD3 pathway. Limiting dilution analysis indicates that accessory cells are not required to mediate the action of IL-6 on CD8+ cells. Furthermore, this action is not blocked by addition of mAb neutralizing either IL-2 or IL2R, thus suggesting that IL-6 does not act via IL-2. CD8+ lymphocytes grown in the presence of PHA + IL-6 incorporate (3H)-thymidine to the same extent as those stimulated with PHA + IL-2, but do not increase in number until day 6 of culture. It is hence apparent that the stimulating activity of IL-6 on CD8+ lymphocytes is restricted to the GO----S phase progression, but does not lead to mitosis. IL-6 receptors are expressed on resting CD4+ and CD8+ lymphocytes: their expression is significantly enhanced on both activated CD4+ and CD8+ cells. Scatchard analysis of (125I)-IL-6 binding data showed the presence of high (Kd, 3 x 10(-10) M) and low (Kd, 6 x 10(-8) M) affinity IL6R on both lymphocyte populations. Similarly, mRNA encoding IL6R was detected in both CD4+ and CD8+ lymphocytes. Thus, our studies indicate that IL-6 directly and selectively stimulates the GO----S progression of CD8+ lymphocytes in the presence of mitogen and absence of IL-2: this phenomenon may be of interest for the elucidation of mechanisms activating cytotoxic T lymphocytes.     

Ability of human T lymphocytes to adhere to vascular endothelial cells and to augment endothelial permeability to macromolecules is linked to their state of post-thymic maturation

The accumulation of mononuclear cells at sites of chronic inflammation is dependent on a number of factors including localized adherence of lymphocytes to vascular endothelial cells (EC), cytokine-mediated increased adhesiveness of endothelium, chemotactic factors and endothelial permeability. The present study investigates two of the above attributes of lymphocyte-EC interaction: namely, the ability of maturationally distinct subpopulations of human T lymphocytes to adhere to vascular EC and to increase vascular endothelial permeability to macromolecules in an in vitro model. Thus, human T lymphocytes were separated into CD4+ CD8-helper/inducer, CD4- CD8+ cytotoxic/suppressor, CD29+ CD45RA- CD45RO+ memory, and CD29- CD45RA+ CD45RO- naive/virgin T subpopulations, were activated with PHA and PMA, and then examined for their adherence to EC and also for their effect on endothelial permeability. Upon activation, cells within each of the above four subpopulations exhibited increased adherence to EC. In contrast, resting CD29+ CD45RA- CD45RO+ memory T lymphocytes exhibited two to three times greater ability to adhere to EC than their CD29- CD45RA+ CD45RO- naive/virgin counterparts. Consistent with their increased adherence to EC, CD29+ CD45RO+ memory T lymphocytes, when activated, significantly increased endothelial permeability to albumin. Although activated CD45RA+ naive T lymphocytes exhibited increased adherence to EC, these cells failed to increase significantly endothelial permeability. Similar to their polyclonal counterparts, Ag-specific CD4+ CD29+ CD45RO+ T cell clones, but not their actively released mediators, also increased endothelial permeability via a noncytolytic mechanism(s). This ability of CD29+ CD45RO+ memory T lymphocytes to augment endothelial permeability may facilitate their transendothelial migration into extravascular space. These observations may provide additional insights into molecular mechanism(s) underlying pathophysiology of localized chronic inflammatory responses in general and more specifically selective accumulation of CD29+/CD45RO+ memory T lymphocytes at sites of chronic inflammation such as rheumatoid synovium.