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Characterization and distribution of lymphocyte subsets in sheep hemal nodes.

Marked differences were found in the percentage and distribution of lymphocyte subsets (CD5, CD4, CD8, T19, CD45, and CD45R cells) in hemal nodes compared to those in mesenteric lymph nodes and blood. There were a greater number of T19 (gamma/delta)+ lymphocytes and fewer CD8+ lymphocytes in hemal nodes than in mesenteric nodes and a higher percentage of CD4+ in hemal and mesenteric lymph nodes than in blood. T19 (gamma/delta)+ lymphocytes were found throughout hemal nodes but were absent from the follicles. Of special interest was the finding that 20% of hemal node lymphocytes appeared not to express CD45 (leucocyte common antigen). According to these results, because of the unique distribution of lymphocyte subsets and the presence of germinal centers, hemal nodes may have a specialized role in the immune system.

Animals↗

Decreased lymphocyte subsets and K-cell activity in iron deficiency anemia.

Lymphocyte subsets and K-cell activity were evaluated in the peripheral blood of 21 patients with iron deficiency anemia. The results showed that the mean number of total lymphocytes, CD3 and CD4 subsets, and B lymphocytes were decreased in these patients. The K-cell activity as measured by specific cytotoxicity and cytotoxic capacity was decreased. However, studies performed in 16 of these patients after the treatment revealed the recovery of these parameters except the decreased K-cell activity. These findings demonstrate immunological abnormalities in iron deficiency anemia which could increase the susceptibility to infections.

Adolescent↗

Determination of lymphocyte subsets and cytokine levels in cynomolgus monkeys.

Lymphocyte subset counts and cytokine assays are useful to investigate the interactions of pharmaceuticals, particularly new biotechnology products, with the immune system. As no specific reagents are available to label monkey lymphocytes or to assay monkey cytokines by ELISA, cross reactivities of a panel of monoclonal antibodies specific for human lymphocytes or cytokines were studied in the Cynomolgus monkey. The proportions of B, T, CD4+ and CD8+ cells were determined by flow cytometry using a whole blood technique with at least one monoclonal antibody for each subset. Background data were obtained for more than 300 samples. Monkey and human cultured white blood cells were stimulated with standard mitogens. PHA + LPS in humans and Con A + PWM in monkeys triggered the greatest proliferation. IL-1 beta IL-2, IL-6, IL-8, TNF-alpha, TNF-beta and IFN-gamma, but not IL-1 alpha, were detected in the monkey using human reagents. In addition, the cytokine profile and the kinetics of cytokine production compared well in humans and Cynomolgus monkeys.

Animals↗

Influence of aging, environmental antigens, and dietary restriction on expression of lymphocyte subsets in germ-free and conventional Lobund-Wistar rats.

Lymphocyte subsets from four groups of Lobund-Wistar (L-W) rats were quantitated to determine the influence of diet restriction (DR) and exposure to environmental antigens on the development of these cells. The following effects were found from 6 to 30 months of age: The number of Ts/c and Ts cells were higher in germ-free (GF) vs conventional (CV) rats, whereas B-cell numbers were lower. W3/13 T cells, Ts, and NK cell numbers were higher in DR vs full-fed rats, whereas B-cell numbers were lower. OX19 and W3/13 T cell numbers decreased from 6 months to 30 months in each group, whereas NK cell numbers increased. Also, OX6+ B cell numbers increased with age, and Ts/c numbers decreased. These data may reflect a relationship between enhanced T-cell function and the extended life span and lower tumor incidence observed in DR L-W rats.

Aging↗

The characterization and function of human immunoregulatory T lymphocyte subsets.

Functional T lymphocyte subpopulations can be identified in humans by antibodies which detect stable glycoprotein antigens on their surface. Thus, inducer T lymphocytes bear an antigen termed T4 while suppressor T lymphocytes bear an antigen termed T5. Immune homeostasis results from a delicate balance between inducer and suppressor subsets within the T-cell circuit and perturbation in subset dynamics may initiate a wide variety of immunopathological disorders. Here Ellis Reinherz and Stuart Schlossman discuss the present understanding of this circuit, its role in the pathogenesis of a number of diseases, and how the human immune response can be manipulated in an orderly way through modulation of selected T-cell subsets.

Antigens↗

[Effects of cordyceps sinensis alcohol extractive on serum interferon-gamma level and splenic T lymphocyte subset in mice with viral myocarditis].

AIM: To investigate the effects of early application of cordyceps sinensis alcohol extractive (CSAE) on myocardial injury, level of serum IFN-gamma and splenic T lymphocyte subset in murine viral myocarditis (VMC). METHODS: 100 male BALB/c mice were divided randomly into control group (CG), infectious group (IG) and CSAE therapy group (CTG). Mice in IG and CTG were infected with Coxsackie virus B3 (CVB3). The 14-day survival rates and myocardial histopathology were observed. Serum IFN-gamma level was detected by ELISA and splenic CD3+, CD4+, CD8+ T lymphocytes were detected by flow cytometry. RESULTS: Serum IFN-gamma level and the percentage of splenic T lymphocyte subset in IG were decreased significantly, and CD4+/CD8+ T cell ratio increased significantly as compared with those of CG. Myocardial lesions in CIG were attenuated, while the 14-day survival of mice 85% (vs IG 55%, P<0.05), serum IFN-gamma and the percentage of splenic CD3+, CD8+ T lymphocytes in CTG were significantly higher than those in IG. There were no significant differences of the percentage of T lymphocyte subset and CD4+/CD8+ T cell ratio between CTG and CG. CONCLUSION: CSAE plays an important protective role against viral-induced murine myocarditis by inducing IFN-gamma and regulating T lymphocyte.

Animals↗

Total numbers of lymphocyte subsets in different lymph node regions of uninfected and SIV-infected macaques.

Human immunodeficiency virus (HIV) infection leads to a decline of CD4+ T-cells in blood. Because blood represents only a small proportion of the total lymphocyte pool, it is important to investigate other lymphoid organs. So far, only relative proportions of lymphocyte subsets in single peripheral lymph node (LN) regions of HIV-infected patients and simian immunodeficiency virus (SIV)-infected macaques have been documented. We have therefore quantified the absolute numbers of lymphocyte subsets in blood and six different LN regions of 10 uninfected and 26 SIV-infected macaques. In addition, we have determined the expression of markers of activation and differentiation. Already, in uninfected monkeys, there were significant differences in the cellular composition of different LN regions. Infection with SIV resulted in drastic changes in the proportion as well as absolute numbers of different lymphocyte subsets. Moreover, the relative contribution of the single LN regions to the total lymphocyte pool was also altered.

Animals↗

Individual lymphocyte subset composition determines cAMP response to isoproterenol in mononuclear cell preparations from peripheral blood.

The aim of the present study was to examine to what extent isoproterenol-stimulated cAMP production in mononuclear cells, isolated from resting human subjects, was correlated with lymphocyte subset composition. Peripheral blood was collected from 14 healthy male subjects, who rested in the sitting position. Mononuclear cells were prepared by density centrifugation and subset composition was determined by two-colour flow cytometry. Cyclic AMP production was determined by a radioimmunoassay after inhibition of phosphodiesterase and stimulation with isoproterenol. NK-cells (CD3-CD56+) were positively correlated to total cAMP concentrations (basal cAMP plus increase in cAMP). Tcytotoxic cells (CD8+) were positively correlated to the relative increase in cAMP. Thelper cells (CD4+) were negatively correlated to total cAMP concentrations and B-lymphocytes (CD(19+20)+) were negatively correlated to the relative increase in cAMP. The results support the view that NK-cells and Tcytotoxic cells have higher beta 2-adrenoceptor densities and more receptors with high affinity than Thelper and B-cells. In conclusion, our study demonstrates that cAMP levels in blood mononuclear cells obtained from normal subjects show substantial variation. This variation is at least in part due to differences in lymphocyte subset composition in the same subjects. Therefore, it is important to know the lymphocyte subset composition when evaluating cAMP accumulation in mononuclear cell preparations from peripheral blood.

1-Methyl-3-isobutylxanthine↗

Circulating immature and mature dendritic cells in relation to lymphocyte subsets in patients with gastrointestinal tract cancer.

Cancer-related deficiency in circulating dendritic cells (DC), whose important anticancer role is well established, has been proven to be associated with lymphocytopenia. This study was performed to evaluate which lymphocyte subset is most markedly related to the failure of the DC system. The study included 30 patients with gastrointestinal tract cancer, 10 of whom had distant organ metastases. Immature and mature DCs were measured by FACS and monoclonal antibodies against CD123 and CD11c antigens, respectively. Low levels of immature and mature DCs were observed in 63% and 43% of patients, respectively. Patients with low levels of circulating mature DCs had significantly lower values of T lymphocytes, T helper lymphocytes and NK cells than those with normal mature DC levels. In contrast, no significant difference was seen between patients with normal or abnormally low values of immature DCs. Conversely, patients with a decreased number of T lymphocytes, T helper lymphocytes and NK cells showed significantly lower values of circulating mature DCs than those with lymphocyte subsets within the normal range, whereas no difference was seen in immature DC amounts. This study suggests that only mature DC deficiency may be associated with important lymphocyte subset alterations in cancer patients, whereas deficiency in immature DCs does not seem to be related to other immune cell disorders.

Antigens, CD↗

[The effect of various occupational exposures to microwave radiation on the concentrations of immunoglobulins and T lymphocyte subsets].

The immunoglobulins' concentrations and T lymphocyte subsets during occupational exposures to microwave radiation were assessed. In the workers of retransmission TV center and center of satellite communications on increased IgG and IgA concentration and decreased count of lymphocytes and T8 cells was found. However, in the radar operators IgM concentration was elevated and a decrease in the total T8 cell count was observed. The different behaviour of examined immunological parameters indicate that the effect of microwave radiation on immune system depends on character of an exposure. Disorders in the immunoglobulins' concentrations and in the T8 cell count did not cause any clinical consequences.

Adult↗

Absolute numbers of lymphocytes subsets migrating through the compartments of the normal and transplanted rat spleen.

Lymphocyte migration is one of the basic principles of the immune system. Up to now lymphocyte migration experiments have been performed either in a quantitative way, determining whole organ recoveries of radiolabeled lymphocytes without histologic localization, or based on autoradiography which does not provide absolute numbers of immigrant lymphocytes. In this study the traffic of lymphocyte subsets through the splenic compartments: red pulp (RP), marginal zone (MZ), periarteriolar lymphatic sheath (PALS) and follicle was evaluated in absolute numbers. In normal spleens and splenic transplants fluorescein isothiocyanate (FITC)-labeled immigrant lymphocytes were localized and characterized immunohistochemically in cryostat sections by light microscopy. In addition morphometry of the splenic compartments was performed and the recovery of 51Cr-labeled lymphocytes in the spleen was determined. The combination of these methods allowed total numbers of immigrant subset cells to be calculated in individual splenic compartments. At 15 min about 17% of the injected B lymphocytes were found in the MZ. This is the largest fraction of an injected lymphocyte subset found in a single splenic compartment. At 24 h immigrant B cells were not only found in the follicle, but they had reached comparable numbers in the three compartments: follicle, RP and MZ. Most immigrant T lymphocytes were found in the PALS, which from 6 h after injection onwards contained more T cell immigrants than any single organ of the body. CD4+ and CD8+ lymphocytes showed a similar distribution throughout the splenic compartments at early time points. At 24 h CD4+ lymphocytes homed preferentially to the PALS, whereas CD8+ cells seemed to prefer the RP and MZ. Both CD4+ and CD8+ cells also migrated into the follicles. In regenerated splenic tissue after autotransplantation lymphocyte immigration was reduced in all compartments and to the MZ in particular. An impaired lymphocyte migration to the MZ in splenic transplants may be one reason for the lack of protection provided against bacterial infections. Thus examining lymphocyte migration in absolute numbers provides additional information which cannot be gained by determining labeling indices or percentages of lymphocyte subsets alone.

Animals↗

Production and culture of HSVtk transduced suicidal lymphocytes induces variable changes in the lymphocyte subset composition.

Allogeneic bone marrow transplant followed by donor lymphocyte infusion (DLI) is limited by T cell-mediated graft-versus-host disease (GVHD). A potential solution to alleviate uncontrolled GVHD is to create a controllable suicidal lymphocyte using retroviral transduction of the herpes simplex virus thymidine kinase gene (HSVtk) into the T cell graft. Should GVHD arise, the administration of ganciclovir (GCV) should eliminate the causal T cells. The culture conditions and expansion protocols required to produce suicidal lymphocytes may affect the composition of the T cell product. In this report we describe how T cells from individual donor samples respond to the same culture condition in highly varied ways. Among five donors, two demonstrated predominant expansion of CD4+ cells (with a decrease of CD8+), two donors resulted in predominately CD8+ cells, and one donor developed mainly dual positive CD8+/CD56+ cells. We observed a 20-fold expansion of T cells during the 14 day protocol. The function of the T cells was not affected by the transduction procedures (as tested by 51Cr release assays). In contrast to suicidal lymphocytes prepared using entire T cell populations, T cells pre-selected into CD3+/CD4+ or CD3+/CD8+ subpopulations prior to culture maintained their initial phenotype during the 14 day culture period, with little or no drift. Results from clinical trials using suicidal lymphocytes may be confounded by variance in lymphocyte subset compositions (LSC) and optimal use of suicidal lymphocytes may require separate culture and transduction to control the LSC delivered to the patient.

Antigens, CD↗

Lymphocyte subsets in a large cohort of patients with systemic lupus erythematosus.

In search of markers of disease activity in patients with SLE we have investigated blood lymphocyte subsets from a large cohort of patient. Seventy-one patients were studied using a well-defined panel of fluorescent monoclonal antibodies which recognize the major T, B and NK lymphocyte subsets and activated cells. Flow cytometry was used with standard automated software. Overall, SLE patients were lymphopenic. The proportion of activated T cells was increased and NK cells were decreased in both proportion and absolute numbers (P < 0.001). This decrease was more pronounced in the more active patients. None of the T cell activation markers was shown to distinguish different degrees of disease activity. However, the percentage of NK cells was significantly reduced in active disease states (P < 0.01). Decreased numbers of NK cells could potentially reduce the resistance of SLE patients to infectious organisms.

Adult↗

Observation of changes in peripheral T-lymphocyte subsets by flow cytometry in patients with liver cancer treated with radioimmunotherapy.

An analysis of lymphocyte subsets was performed in 24 patients with surgically verified unresectable hepatocellular carcinoma (HCC). In all cases, the hepatic artery was cannulated and ligated and a single dose of 131I-Hepama-1 monoclonal antibody (MAb) was infused through a hepatic artery catheter. Blood samples were obtained before infusion and 1 month post-infusion. The results indicated that the CD3+ T-cells (pan T-cells) were normal, whereas the CD4+ T-cells (T-helper/inducer cells) decreased and the CD8+ T-cells (T-suppressor/cytotoxic cells) increased significantly in comparison with the control group. So the CD4+:CD8+ ratio declined markedly. One month post-radioimmunotherapy (RIT), the T-cell subsets did not change, but CD4+, CD8+ and the CD4+:CD8+ ratio differed significantly between patients with and without sequential resection. The changes in the T-cell subsets were closely related to tumour capsule status and the human anti-murine antibody (HAMA) response, but no relationship was found between the tumour and administration of a moderate dose of radioimmunoconjugate. Therefore, determination of changes in the T-lymphocyte subsets and the CD4+:CD8+ ratio could be used as clinical and prognostic indicators in patients who have received RIT. Furthermore, RIT did not affect the distribution of T-cell subsets in patients with HCC.

Animals↗

Lymphocytes and T lymphocyte subsets are regionally distributed in the female goat reproductive tract: influence of the stage of the oestrous cycle.

The reproductive tract of the female is a part of the mucosal system which protects from pathogens invasion. We have analysed the presence and distribution of total lymphocytes, plasma cells (antibody secreting B cells) and T lymphocytes subsets in the reproductive tract of the female goat. The influence of the oestrous cycle on the densities of lymphocytes and plasma cells of the cervix and uterus horn was evaluated in sections prepared for conventional histology. Immunocytochemistry was used for the study of lymphocyte subsets by confocal microscopy and immunoperoxidase techniques. Present results show that the reproductive tract of the goat is a site rich in lymphocytes. These cells were found mingled with the epithelial cells of the endometrium and distributed throughout the stroma. Lymphocyte aggregates were observed in the stroma. Lymphocyte but not plasma cell number changed depending on the reproductive stage of the goats. The impact of the hormonal environment was different for the cervix and uterine horn. Immunocytochemistry studies evidenced the presence of cells displaying immunoreactivity for both CD 4+ and CD 8+ antibodies in the epithelial layer and stroma of the cervix and uterine horn. These cells were more numerous in the cervix and were also found infiltrating the luminal epithelia of endometrial glands. Overall, our results indicate that lymphocyte distribution is different in the cervix and the horn, and is influenced by the stage of the reproductive cycle. In summary, CD 4+ and CD 8+ T lymphocytes subsets could be found in the endometrium of both the cervix and uterine horn of the goat reproductive tract.

Animals↗

Influence of feeding formula and breast milk fortifier on lymphocyte subsets in very low birth weight premature newborns.

BACKGROUND: It is well known that breast-feeding protects the newborn from infectious diseases. This is especially important for very low birth weight preterm infants, whose immune systems are immature. In this study we investigated how a milk fortifier and replacement formula affected lymphocyte subsets in preterm infants. METHOD: The study assessed the effects of different types of feeding (human milk, n = 14; fortified human milk, n = 16; formula, n = 14) on lymphocyte subsets in 44 very low birth weight preterm infants. For each baby, two consecutive blood samples were collected 7-10 days apart during the full enteral feeding period. For each sample, the percentages of CD3+ (pan-T), CD19+ (B-cell), CD4+ (T-helper), CD8+ (T-suppressor), and CD3-CD16/56+ (natural killer cell) lymphocytes were measured in a flow cytometer, and the absolute count for each subset was calculated based on the total lymphocyte count. Within each feeding group, the absolute numbers of each lymphocyte subset in the two consecutive samples were compared. Also, the mean absolute counts for each cell type were compared among the 3 groups for the first set of blood samples, and the same comparisons were made for the second set. RESULTS: The mean number of CD3-CD16/56+ cells in the formula-fed infants was significantly lower than the corresponding means in the groups fed human milk alone and fortified human milk (p = 0.037). CONCLUSION: The findings suggest that babies fed formula have different lymphocyte subset compositions than those fed breast milk or fortified breast milk.

B-Lymphocytes↗

The effect of standardized ginseng extract on peripheral blood leukocytes and lymphocyte subsets: a preliminary study in young health adults.

The present study was undertaken in twenty young, healthy Thai males to investigate the effects of prolonged administration of standardized ginseng extract on peripheral blood leukocytes and lymphocyte subsets. The subjects were divided into two equal groups as ginseng and placebo groups. The first group received two capsules daily of standardized ginseng extract 150 mg per capsule for 8 weeks. The second group received placebo and served as control. Circulatory levels of total and differential leukocyte counts and percentage of lymphocyte subsets were determined prior to and at the 4th and 8th week of the experimental period. There were no significant differences in the total and differential leukocyte counts as well as the lymphocyte subpopulations: CD3 (T cells), CD 19 (B cells), CD4 (T-helper cells), CD8 (T-suppression cells), CD4/CD8 ratio, and CD25 (Interleukin-2-receptor cells) between the two subject groups throughout the experimental period. We concluded that oral administration of standardized ginseng extract, 300 mg/day for 8 weeks caused no significant changes in peripheral blood leukocytes and lymphocyte subsets in young, healthy Thai males.

Administration, Oral↗

Lymphocyte subsets in pulmonary venous and arterial blood of lung cancer patients.

Lymphocyte subsets in pulmonary venous blood (PVB) smears from 42 patients with lung cancer were immunocytochemically determined. In four patients, pulmonary arterial blood (PAB) smears were also studied for comparison with PVB. Seven healthy donor peripheral blood (PB) smears were used as controls. The percentage of T cells, helper/inducer T (Th) cells and B cells were significantly lower (P less than 0.01) than in normal controls but those of suppressor/cytotoxic T (Ts) cells, natural killer (NK) cells (P less than 0.01) and S100+ small lymphoid cells (P less than 0.05) were higher. This resulted in a decrease in the Th:Ts value in cancer patients (1.46 vs. 2.28 for normal controls; P less than 0.01). The Th and Ts value of PVB from patients in pathological Stages III and IV was lower than from those in Stages I and II because of the increase in Ts cells in the former (P less than 0.05). S100+ small lymphoid cells were increased in cancer patients, especially in those with adenocarcinoma. The present study demonstrates immunoregulation abnormalities in cancer bearing hosts, the results correlating well with the stage of the cancer. Determining lymphocyte subset alterations in PVB did not, however, enable us to detect the changes associated with local immune responses against cancer.

Adult↗