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Effects of regular exercise on lymphocyte subsets and CD62L after psychological vs. physical stress.

OBJECTIVE: To examine the effects of regular physical activity on lymphocyte responses to a speech stressor and an exercise challenge. METHODS: We assessed lymphocyte subsets and CD62L expression pre, immediately after and 15 min after a speech task vs. exercise in 24 high vs. 24 low physically active subjects. Catecholamine levels were determined by radioenzymatic assay, and enumeration of cells was assessed by flow cytometry. RESULTS: Both tasks induced significant increases in plasma epinephrine (EPI; P<.05) and norepinephrine (NE; P<.001) levels. Similarly, both tasks led to increases in the numbers of lymphocyte subsets (P<.05). Physically active individuals showed attenuated responses to the speech stressor in numbers of CD62L(+), CD45RA(+), CD45RO(+) CD8(+), CD45RO(+) T(H) and CD62L(-) natural killer (NK) cells (P's<.05). In contrast, physical activity level had no significant effect on lymphocyte subsets or CD62L expression in response to exercise. CONCLUSION: The findings suggest that physical fitness affects immune responses to a psychological but not a physical stressor. It is an interesting but open question whether attenuated lymphocyte trafficking responses to stress in regular exercisers might have clinical implications regarding host defense by the immune system.

Adult↗

Peripheral blood lymphocyte subsets and survival in small-cell lung cancer.

Measurement of lymphocyte subsets in peripheral blood is likely to reflect immune response of patients and therefore may be linked to survival in small-cell lung cancer (SCLC). Forty patients with SCLC (14 with limited and 26 with extensive disease at study entry) were included and followed for up to 23 months. Peripheral blood lymphocytes were determined by flow cytometry and levels for total-, T-, B-, helper-, suppressor-, activated T-cells and natural killer cells established and the T-helper-suppressor ratio (H-S ratio) calculated. Quartiles of lymphocyte subset cell counts at the start of follow-up and for the change over follow-up were investigated. Cox regression models indicated that H-S ratio was a significant predictor (p = 0.02) of survival. The risk ratio, after adjustment for competing risk factors, sex, age and stage of disease, was found to be 0.66 (95 percent confidence intervals, 0.46 to 0.95) and predicted that for each quartile of greater H-S ratio, the risk of death decreased by 34 percent. For change in cell counts over follow-up, total lymphocytes (p = 0.02) demonstrated a significant association with survival. Although our observation is limited by the fact that 13 patients were unavailable for follow-up, we conclude that H-S ratio can serve as an easily accessible marker of immune function and possible prognostic value in SCLC.

Carcinoma, Small Cell↗

Changes in lymphocyte subset distribution aid in the differential diagnosis of renal allograft dysfunction.

The distribution of selected lymphocyte subset populations in renal transplant patients was used to assist in the differential diagnosis of graft dysfunction. Patients experiencing dysfunction due to rejection showed consistent and significant decreases in relative numbers of CD 8 (cytotoxic/suppressor) lymphocytes. The ratio of CD 4 cells to CD 8 cells in this group of patients was generally greater than 2.00 due to decreases in CD 8 cells. Patients showing graft dysfunction due to viral infections showed consistent and significant increases in CD 8 cells which also bear the HNK-1 or the HLA-Dr determinants. Serial monitoring for these dual-marked lymphocytes on a weekly basis can be of considerable use in determining the etiology of graft dysfunction. Increases in other "activation" markers, including transferrin receptors, CD 38, and a T cell lineage specific activation antigen (TLiSA) were not specific for rejection; in fact, increases in CD38 were more often associated with viral infections. These studies indicated that lymphocyte subset determinations done on a regular basis can help distinguish graft dysfunction due to viral infections from other causes. The ability to distinguish rejection episodes from stable grafts is less obvious. Although the alterations in lymphocyte subset distribution are not entirely specific, they can distinguish viral infections from rejection.

Diagnosis, Differential↗

Induction of immunoglobulin-secreting cells in the allogeneic mixed leukocyte reaction: regulation by helper and suppressor lymphocyte subsets in man.

The existence of functionally distinct T lymphocyte subsets in man, initially demonstrated with heteroantisera, has been confirmed with monoclonal reagents. Two major subsets have been defined: Leu-2 cells, which effect cell mediated lymphocytotoxicity (CML), and Leu-3 cells, which amplify CML and other T cell functions. This study is an effort to determine the effects of these subsets on the immunoglobulin secretion induced in the human mixed leukocyte reaction (MLR). T cells were separated from non-T cells by rosetting with sheep erythrocytes and were fractionated into Leu-2 and Leu-3 subsets by solid phase immunoabsorption with monoclonal antibodies. T cell subsets were cultured with autologous non-T cells and irradiated allogeneic stimulator non-T cells. Secretion of IgM and IgG was measured by a reverse hemolytic plaque assay. MLR-induced antibody secretion was specifically dependent on the Leu-3 T lymphocyte subset. The Leu-2 subset was incapable of generating large numbers of IgM- or IgG-secreting cells, and, in fact, suppressed the Leu-3-induced response. Exposure of Leu-3 cells to a dose of irradiation sufficient to prevent their proliferation in MLR did not reduce induced immunoglobulin secretion. Leu-2-mediated suppression, however, was sensitive to low dose irradiation. Thus Ig secretion in human MLR is regulated by a balance of helper activity from the Leu-3 subset and suppressor activity from the Leu-2 subset.

Antibody-Producing Cells↗

Lymphocyte subset profile of young healthy children residing in a rural area: possible role of recurrent gastrointestinal infections.

BACKGROUND: Lymphocyte subsets in healthy children are currently characterized by age-related standards. Because antigenic stimuli play a role in maturation of the immune system after birth, there is a question of whether cellular immune development differs in infants whose living conditions entail extensive antigenic exposure and infants growing up in a more protected environment. METHODS: Peripheral blood lymphocyte subsets were studied in two populations of children of similar age and nutritional status; children belonging to a rural population residing in proximity with farm animals and children from an economically privileged urban population. In each population, children studied included a group with an acute diarrheal episode and a healthy control group. RESULTS: Among rural population children, 65% had experienced at least one episode of gastroenteritis within the previous 3-month-period, compared with less than 10% of urban population children. In the rural population group 15% had experienced two or more episodes of gastroenteritis. The proportion of helper T cells was similar in rural population and urban population children. Among helper T cells, the proportion of CD29+ "memory" cells of the total CD4+ helper T cells was more than two times higher than those in rural population children. The proportion of CD8 cells was higher in rural population children than in urban population children, and the proportion of natural killer cells, CD56+ and CD57+, was two to three times higher in rural population children. Within each population, peripheral blood lymphocyte subsets did not differ between the healthy control group and those with acute diarrhea. CONCLUSIONS: In young children exposure to environmental pathogens and specifically to gastrointestinal antigenic stimuli is a major factor affecting development of the cellular immune response. Young children who have experienced enhanced infectious exposure have a peripheral blood lymphocyte profile similar to that of adults.

Acute Disease↗

[Decidual lymphocyte subsets in pregnant women].

OBJECTIVES: Materno-foetal immunological reactions in decidua are probably one of the most important elements in pathogenesis of preeclampsia. DESIGN: To compare lymphocyte subsets isolated from decidua of preeclamptic pregnant women with lymphocyte subsets isolated from healthy pregnant women. MATERIALS AND METHODS: Preeclampsia (PE) was defined according to USA National Health Institute criteria. The study group consisted of 21 women with PE and 11 women with physiological pregnancy. All pregnancies were finished with elective cesarean sections. Exclusion criteria were: uterine contractions, infection, chorinamnionitis, diabetes mellitus and therapy with steroids less than 7 days before blood sampling. Decidual tissue was obtained by curettage of the uterine cavity. The fragments of decidua were separated from clotted blood and placed in sterile tubes with 5 ml of isotonic solution (PBS). Then the decidual tissue was mechanically fragmented, homogenized and rinsed in PBS. Routine immunofluorescent marking techniques with monoclonal antibodies were performed. Analysis was done with FACSCalibur flow-cytometer with 488 nm argon laser using CellQuest programme. The following lymphocyte subsets were estimated: CD3, CD19, CD4, CD8, CD4/CD29, CD8/CD28, CD4/CD45RA, CD4/CD45RO, CD56/CD16, CD69. The results were described as percentage of lymphocytes positive for above surface molecules. Statistical analysis was performed using t-Student and U-Mann-Whitney tests. The work was sponsored by KBN 4 P05E 118 15 grant. RESULTS: Decidua of pregnant PE women contains significantly increased percentage of CD3-/ CD56 + 16+, CD8+/CD28+ cells and decreased percentage of CD3+, CD19+, CD4+/CD29+ and CD4+/CD45RA+ compared to decidua of healthy pregnant controls. CONCLUSIONS: These changes suggest that deficiency of suppressor activity as well as aberrant immunoregulation exists in decidual tissue of PE women.

Adult↗

Increased prevalence of colonic polyps and altered lymphocyte subset pattern in the colonic lamina propria in acromegaly.

OBJECTIVE: The balance of evidence suggests that acromegaly is a risk factor for colonic neoplasia. We have evaluated the prevalence of colonic polyps in acromegalics from Southern Italy and characterized the lymphocyte subsets in the colonic lamina propria in order to analyze differences in the colonic immunological environment. DESIGN: All the patients and controls were submitted to pancolonoscopy. Ten per-endoscopic biopsies of the intestinal mucosa surrounding polyps were carried out to evaluate lymphocyte subsets. PATIENTS: Fifty acromegalics and 318 sex- and age-matched controls entered this study. Colonic lamina propria lymphocyte subsets were studied in 34 patients and 34 controls. RESULTS: Colonic polyps were resected in 23 acromegalics (46%) and 42 controls (13.2%; P < 0.0001); hyperplastic polyps were found in 24% and 6.3%, adenomatous polyps in 22 and 6.9%, (P < 0.01), adenocarcinoma in 2 and 1.2% while synchronous polyps occurred in 18% and 2.5% (P < 0.01), respectively. The number of polyps was significantly correlated with age both in acromegalics (r = 0.422, P < 0.005) and in controls (r = 0.865, P < 0.001). However, polyp prevalence was greater in patients aged below 40 yrs (r.r = 1.9) and in patients with two or more skin tags (r.r = 1.2). A significant decrease of CD20, CD19, CD16, gamma/delta, CD4@leu8- and increase of CD3 and CD4+/leu8+ was found in the lamina propria lymphocyte subsets. CONCLUSIONS: The results of this study confirm that acromegalics are at increased risk of colonic polyps compared to the healthy population. The increased prevalence of premalignant polyps, namely the adenomatous type, suggests that acromegalics should undergo a careful screening and follow-up by pancolonoscopy. An impairment of mucosal immune surveillance seems to exist in acromegaly although a causal effect in the polyp formation cannot be ruled out.

Acromegaly↗

Are the changes of lymphocyte subsets in sickle cell anemia due to the loss of splenic function?

Lymphocyte subsets and K cell activity were evaluated in sickle cell anemia (SCA) and in splenectomized patients. Results showed that the number of total lymphocytes, T lymphocyte subsets and B lymphocytes were increased in SCA. However, individuals who had undergone splenectomy did not exhibit all these abnormalities, suggesting that the lack of the spleen apparently was not the unique factor responsible for the lymphocyte abnormalities seen in SCA patients.

Adolescent↗

T-lymphocyte subsets in nephrotic syndrome.

T-lymphocyte subsets when measured in steroid responsive nephrotic syndrome (SRNS) have demonstrated significant variance from normal values. T-cell subsets were studied by using two-color flow cytometric analysis in 32 children (9.2 +/- 5 years of age) with SRNS. The children were divided into four groups: a) SRNS in acute relapse, on prednisone; b) SRNS in acute relapse, off prednisone; c) SRNS in long-term remission, off prednisone (nephrotic controls); d) patients in remission on long-term prednisone therapy; and e) 15 age-matched normal controls. Children suffering an acute relapse of SRNS showed an increase in Leu2a+/DR+ (CD8) activated lymphocytes (P less than 0.05), a decrease in Leu4a+ total T-lymphocytes (P = 0.01) and a decrease in Leu3a+ (CD4) helper T-cells (P less than 0.05) when compared to normal controls and nephrotic controls. Though some subset changes may represent a prednisone effect and the functional role of these lymphocytes in the disease process is unknown, this study provides additional evidence to support a role for abnormal T-cell subsets in the etiology of SRNS.

Adolescent↗

Absence of effects of cyclosporine on myocardial lymphocyte subsets in Coxsackievirus B3 myocarditis in the aviremic stage.

To test the therapeutic efficacy of immunosuppression with cyclosporine upon the aviremic stage of coxsackievirus B3 (CB3) myocarditis, 2-week-old BALB/c mice were inoculated with 3 x 10(2) plaque-forming units of CB3, and the effects of cyclosporine on peripheral, splenic, and myocardial lymphocyte subsets were investigated. Cyclosporine, 25 mg/kg/day, was administered subcutaneously daily on days 10-31 (experiment 1) and days 30-51 (experiment 2). Treated groups were compared with infected controls for each experiment. In experiment 1, the survival rate of the cyclosporine-treated group was low (17/25 vs. 24/25, p less than 0.05). The severity of myocardial lesions and the distribution of lymphocyte subsets in myocardium and spleen on days 15-18 did not differ between treated and control groups; on the other hand, the percentages of peripheral Thy 1.2+ (pan T) and L3T4+ (activated helper T) cells on days 15-18 were decreased in the treated group, and those of B, Lyt 1+ (helper/inducer T), and Lyt 2+ (suppressor/cytotoxic T) subsets did not differ significantly. Notably, myocardial interleukin-2 receptor (IL-2R) positive cells, through which cyclosporine is considered to act, were scarce in both groups. In experiment 2, survival rates of two groups did not differ (treated, 32/34; untreated, 34/34; p = NS). The severity of myocardial lesions and the distribution of splenic lymphocyte subsets on days 35-38 also were not different between two groups. The percentages of peripheral lymphocyte subsets (Thy 1.2+ and L3T4+) were decreased in the treated group; those of B, Lyt 1+, and Lyt 2+ subsets did not differ significantly. In experiments 1 and 2, the thymus/body weight ratio in the treated groups was smaller than in the untreated group, but the spleen/body weight ratio in the treated group did not differ from the untreated group; histologically, medullary cellular depletion was evident in the thymus, not in the spleen, of the treated groups. We conclude that cyclosporine failed to change the distribution of lymphocyte subsets in the spleen as well as in the myocardium in CB3 myocarditis although it had effects on the peripheral blood and thymus, which may account for the higher mortality in the treated groups. The absence of beneficial effects of cyclosporine upon the CB3-infected myocardium may be related to the paucity of cyclosporine-sensitive cells (IL-2R, L3T4, and Lyt 2 positive cells) in the myocardium.

Animals↗

Enumeration of absolute numbers of T lymphocyte subsets in B-chronic lymphocytic leukaemia using an immunoperoxidase technique: relation to clinical stage.

An immunoperoxidase technique has been used to identify and enumerate helper and suppressor T-cell subsets, as defined by reactivity with Coulter T4 and OKT8 monoclonal antibodies in 54 patients with B chronic lymphocytic leukaemia (B-CLL) and in the same number of matched controls. The ratio of T4+ to T8+ cells was significantly reduced in the B-CLL group as a whole (P less than 0.001) and in each stage of the three clinical staging systems. There was an increase in the median absolute level of T8+ cells in the whole CLL group (P less than 0.001). However, subdivision of the CLL group by clinical staging systems revealed a large group (28 patients) in which median T8+ cell levels were not raised and median T4+ cell levels were low (P less than 0.01). There was no significant decrease in T4+:T8+ ratio, increase in T8+ cells or decrease in T4+ cells with progression of clinical stage. Absolute numbers of E+ cells were significantly raised in all staging systems as were E+ T4- T8- cells (P less than 0.001). A significant alteration in either of these populations with progression of clinical stage was not present.

Adult↗

Monitoring of T lymphocyte subset during ATG induction therapy in hand allograft with report of 3 cases.

OBJECTIVE: To investigate the significance of T lymphocyte subset determination during antithymocyte globulin (ATG) induction therapy in reducing the total drug dose, incidence of complications and cost of treatment in hand allograft. METHODS: The changes in peripheral blood T lymphocyte subsets (CD3+, CD4+, CD8+, and CD28) were determined by flow cytometry in 3 cases of hand allograft who received ATG treatment. RESULTS: Flow cytometry showed that the percentages of CD3+, CD4+, and CD8+ T lymphocytes, along with the ratio of CD4/CD8, decreased significantly during ATG induction therapy, and the results were consistent in the 3 cases. Long-term continuous changes of peripheral blood lymphocytes were observed after antithymocyte globulin induction therapy. CONCLUSION: The understanding of the immunological state of the patient with hand allograft after ATG induction therapy by monitoring T lymphocyte subsets may allow adjustment of the total dose of the drugs administered and help prevent the occurrence of complications.

Adult↗

Physiological changes of Fas expression in peripheral lymphocyte subsets during the menstrual cycle.

We have examined changes in peripheral lymphocyte subsets, and Fas expression in these subsets, during the menstrual cycle. Measurements were made by three-color flow cytometry in the follicular and luteal phases of the menstrual cycle in ten healthy women. The numbers of leukocytes, granulocytes and monocytes were significantly higher in the luteal phase than the follicular phase. The percentage of CD8(+) cells was greater in the luteal phase than the follicular phase. The percentages of Fas(+) cells among T cells and NK cells were higher in the luteal phase than the follicular phase. These findings suggest that the menstrual cycle affects leukocytes, lymphocyte subsets, and Fas expression in these subsets, and that changes in the luteal phase of the menstrual cycle are similar to those in pregnancy.

Adult↗

[T lymphocyte subsets in aqueous humor from patients with uveitis].

We studied T lymphocyte subsets in the aqueous humor (AH) and peripheral blood (PB) from patients with active uveitis using three-color flow cytometry. We assessed the number of T lymphocyte subsets that stained with fluorescence-conjugated anti-CD3, CD4, CD8, CD29, CD45RA, CD45RO, and HLA (human leukocyte antigen)-DR monoclonal antibodies in the AH and PB from 32 patients: 9 with sarcoidosis (SAR), 6 with Vogt-Koyanagi-Harade disease (VKH), 4 with HLA-B27+ uveitis, 2 with Behçet's disease, 2 with diabetic iritis, 9 with idiopathic uveitis, and 10 healthy control subjects. The percentage of CD3+ lymphocytes was significantly higher in uveitic AH than in PB. The percentage of CD4+ lymphocytes was significantly higher in uveitic AH than in PB in SAR. While the percentage of CD4+CD45RA+ (naive) cells within T cells was much lower in uveitic AH than in PB, the percentage of CD4+CD29+ or CD4+CD45RO+ (memory) cells was significantly higher. There was no significant difference in these subsets in PB between patients and controls. The results suggest that an increase in the number of activated memory T lymphocytes in AH is involved in the pathogenesis of uveitis.

Adult↗

Oral pulse prednisone therapy after relapse of severe autoimmune chronic active hepatitis. A prospective randomized treatment trial evaluating clinical, biochemical, and lymphocyte subset responses.

To assess the efficacy of oral pulse prednisone therapy after relapse of severe autoimmune chronic active hepatitis and to determine the effects of such therapy on serum immunoglobulin G levels and peripheral blood lymphocyte subsets, 16 patients were randomized after relapse to therapy with prednisone 90 mg daily for 5 days each month or conventional daily dose therapy with prednisone in combination with azathioprine. Seven of the eight patients randomized to pulse therapy failed treatment after 1.9 +/- 0.7 months (range, 24 days to 6 months). Treatment failure occurred more commonly (87 vs. 0%, p < 0.01) and remission occurred less frequently (0 vs. 87%, p < 0.01) in patients receiving pulse therapy. The percentage and absolute numbers of circulating CD2 and CD4 cells decreased during relapse, while CD8 and CD20 cell counts were not significantly altered. Pulse therapy was associated with a significant increase in the percentage of CD4 cells and decrease in aminotransferase levels after 5 days. Serum immunoglobulin G levels, however, were not affected by treatment and after 1 month the biochemical findings and lymphocyte subsets were again similar to those at relapse. In contrast, serum immunoglobulin G levels decreased after 14 days of combination therapy and by 1 month the numbers in all lymphocyte subsets were higher than at relapse. We conclude that oral pulse prednisone therapy is ineffective for patients in relapse. The numbers of circulating CD2 and CD4 cells fall during relapse and increase as inflammatory activity subsides during corticosteroid therapy. Combination therapy reverses the biochemical changes and alterations in peripheral blood lymphocyte subsets that accompany relapse.

Administration, Oral↗

Study of hematopoietic progenitor cells, hematological values and lymphocyte subsets in cord blood: application for cord blood transplantation.

Hematological values, lymphocyte subsets and hematopoietic progenitor cells from normal term cord blood samples were studied, compared with normal adult blood, and analysed to determine whether a single collection of cord blood is sufficient for transplantation in adults. The parameters were assayed by automatic cells counter, flow cytometry and semisolid cell culture. All of the hematological values except RBC and MCHC were higher than in normal adult blood. Sex had an influence on RBC, Hb, Hct, Plt and reticulocyte counts. For lymphocyte subsets, all of the absolute CD3+, CD4+, CD8+ counts and T helper: suppressor ratio were higher than those of adult blood. All of the hematopoietic progenitor cells in cord blood were also higher than in adult blood. The mean volume of cord blood for each collection was 80.75 +/- 4.81 ml and the mean numbers of nucleated cells, CFU-GM and CD34+ were 13.51 +/- 0.38 x 10(8) cells, 4.33 +/- 0.66 x 10(5) colonies and 42.65 +/- 7.00 x 10(5) cells respectively. This 80 ml of cord blood would contain sufficient marrow repopulating cells for a recipient weighing about 20 kg. Recently developed technology, including ex vivo expansion may even permit transplants in adults.

Adult↗

[Role of Fas-FasL and caspase-3 signal transduction pathway in promoting apoptosis of T lymphocyte subset in SLE patients].

AIM: To explore the relationship between Fas-FasL-mediated signal transduction pathway and apoptosis disorder of T lymphocyte subset in SLE patients. METHODS: The expression rate of membrane Fas, FasL and that of intracellular activated caspase-3 of T lymphocyte subset were determined by flow cytometry. RESULTS: Compared with healthy control group, the expression rate of membrane Fas on CD4(+) T cells significantly increased in SLE patients in the active and inactive phases (P<0.05), however, that on CD8(+) T cells slightly increased but there was no statistical significance (P>0.05). The expression rate of FasL on T cell subset in SLE patients in the active and inactive phases significantly increased (P<0.05) but there was no obvious difference of the expression rate of Fas and FasL on T cell subset between the two disease groups (P>0.05). The expression rate of intracellular activated caspase-3 in T cell subset of SLE patients in the active phase was notably higher than that in the inactive phase and healthy control group (P<0.05). The expression rate of intracellular activated caspase-3 in T cell subset of SLE patients in the inactive phase was slightly higher than that in health control group but there was no statistical significance. CONCLUSION: Apoptotic speed of T lymphocyte subset in SLE patients was accelerated while CD4(+) T cells were in a state of active apoptosis. It is possible that Fas-FasL signal transduction pathway plays an important role in the induction of T cell apoptosis. The degree of apoptosis of T lymphocytes closely correlates with the disease's activity in SLE patients.

Adolescent↗

Determination of CD30 expression on peripheral blood T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome by FCM.

To determine the CD30 expression on peripheral blood T lymphocyte subsets in patients with hemorrhagic fever with renal syndrome (HFRS) and its clinical implications, double immunofluorescence technique and flow cytometry were used. There was no significant difference among the severe group, mild-moderate group and normal control group in the CD4+CD30- T lymphocyte subset. While the CD4+CD30+ T cells of HFRS patients were increased and the difference between severe group and mild-moderate group or normal control group were very significant (P < 0.01) and the difference between the mild-moderate group and normal control group was also significant (P < 0.05). The CD8+CD30- T cells were increased while the CD8+CD30+ T cells decreased obviously in HFRS patients, and the differences among three groups in both subsets were very significant (P < 0.01). The results showed that the humoral immunity and cellular immunity are overactive in HFRS patients during acute phase. The loss of balance between T lymphocyte subsets may play an important role in the pathophysiology of HFRS and is closely correlated with the severity of the HFRS.

Adult↗