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PubMed1995

[The change of lymphocyte subset and effect of FK506 in experimental small bowel transplantation of rats].

I analyzed the relationship of the change of lymphocyte subsets in the blood and acute allograft rejection of heterotopic small bowel transplantation in the rats. The effect of immunosuppressive agent, FK506 was monitored by lymphocyte subsets and effective dose in small bowel transplantation was assessed. Four different combinations were investigated: (A) syngenic graft; (B) allogenic graft without FK506; (C) allogenic graft with FK 0.135mg/kg/day; and (C-2) allogenic graft with FK 0.270mg/kg/day. Lymphocytes obtained from the caudal artery blood at 1, 3, 5 and 7 days after transplantation, were stained by monoclonal antibodies (W3/13, W3/25, OX-8, 68-IB3) and then enumerated by flow cytometry. In group A, lymphocyte subsets showed no significant change after transplantation. Group B showed remarkable decrease of W3/13 and OX-8 positive cells at 3 and 5 days after transplantation. In group C-1, the change of lymphocyte subsets was similar to group B. In group C-2, the lymphocyte subsets revealed almost same change as group A. The change of lymphocyte subsets, especially W3/13 and OX-8 positive cells, may be useful marker for early diagnosis of acute rejection and for determination of the effective dose of FK506 in the small bowel transplantation.

Animals↗
PubMed1993

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.

Adult↗
PubMed1999

Quantification of proliferating lymphocyte subsets appearing in the intestinal lymph and the blood.

Lymphocyte emigration from the intestinal wall via lymphatics is necessary to maintain gastrointestinal immunity and also to connect the different parts of the mucosal immune system. In the present study the numbers and time kinetics of proliferating lymphocyte subsets leaving the gut wall via intestinal lymphatics were analysed in mesenteric lymph node adenectomized minipigs (n = 8). After cannulation of the major intestinal lymph duct, afferent lymph was collected under non-restraining conditions. In four pigs lymphocytes taken from the intestinal lymph and blood were incubated in vitro with the thymidine analogue bromodesoxyuridine (BrdU) to label all lymphocytes in the S-phase of the cell cycle. The other four pigs received a single i.v. injection of BrdU 1 week after cannulation. The initial percentage of BrdU+ lymphocyte subsets in the intestinal lymph 15 min after BrdU injection was comparable to that after the in vitro labelling (1.5 +/- 0.7% in T cells, 10.6 +/- 1.6% in IgM+ cells and 30.0 +/- 11.9% in IgA+ cells). From this level onwards, the percentage of in vivo labelled BrdU+ lymphocyte subsets reached a maximum at 12 h after BrdU application. A different pattern of BrdU+ subsets was seen in the blood. After an early peak at around 3-4 h, the frequency of BrdU in vivo labelled cells decreased. Each subset had a maximum between 12 h and 48 h after BrdU application (maximum of BrdU+ CD2+ T cells at 12 h, 4.6 +/- 1.5%; IgM+ BrdU+ at 48 h, 8.8 +/- 3.3%). The present results provide a basis to determine the time necessary for induction of specific intestinal immunity during oral vaccination studies.

Animals↗
PubMed1986

Lymphocyte subsets in patients with dilated cardiomyopathy and perimyocarditis.

Lymphocyte subsets were analyzed in patients with DCM, arrhythmias following influenza symptoms and acute perimyocarditis, respectively by an automated laser flow cytometry system with the use of monoclonal antibodies. A lower percentage of OKT8+ cells and a high OKT4/OKT8 ratio were found in the DCM and arrhythmias groups. No phenotypic abnormalities in the lymphocyte subsets were found in patients with acute perimyocarditis who were favorably progressing in clinical features. The results suggest that an immunological disturbance thought to play a part of the pathogenesis of DCM or heart failure may cause the abnormality of lymphocyte subsets.

Adult↗
PubMed1997

Multivariate reconstruction of lymphocyte profiles in a two-dimensional graphical model as a tool for the investigation of lymphocyte subset distribution in health and disease.

Advanced multivariate data-analytical techniques are proposed to concisely represent and evaluate complex lymphocyte profiles (i.e., compound lymphocyte subset distributions) of individual subjects in easily interpretable, two-dimensional, graphical correlation biplots. The lymphocyte profile of each subject is represented by its location in the model, and the score of a subject for a particular lymphocyte subset is inferred from the perpendicular projection on a rotated axis that coincides with this lymphocyte subset. Simultaneously, the model yields information about the correlation between the lymphocyte subsets. Furthermore, individuals with aberrant lymphocyte profiles can be easily identified. In case studies of 80 healthy donors and of 40 patients with multiple myeloma, 10 patients with monoclonal gammopathy of undetermined significance, and 50 age- and sex-matched healthy donors, reconstruction of the two-dimensional lymphocyte profiles from 27 flow-cytometric characterized lymphocyte subsets succeeded in representing 43% and 51% of the total information (variability) contained within the 80 x 27 (= 2,160) and 100 x 27 (= 2,700) flow cytometry measurements, respectively. It is concluded from the present studies that the correlation biplot represents a unique and powerful tool to concisely describe, represent, and analyze complex lymphocyte profiles of individual subjects and the heterogeneity in lymphocyte profiles among these subjects.

Adult↗
PubMed1987

Peripheral blood lymphocyte subsets in nasopharyngeal carcinoma.

Peripheral blood lymphocyte subsets were studied in 29 untreated Chinese nasopharyngeal carcinoma (NPC) patients and 20 normal controls of similar age range and sex ratio with indirect immunofluorescence using monoclonal antibodies. The percentage T lymphocyte (T11) count, percentage and absolute T helper (T4) lymphocyte counts and the T helper/T suppressor-cytotoxic cell ratio (T4/T8) were significantly lower, while the percentage and absolute T8 counts were significantly higher in NPC patients. Concerning B lymphocytes, B1 cell count was normal while B4 cell count (B lymphocytes including early B lymphocytes) was significantly lower in NPC patients. These various lymphocyte subset changes were not related to the clinical stage or to Epstein-Barr viral IgA viral capsid antibodies and IgA early antibodies. The pathogenetic and prognostic significance of peripheral blood lymphocyte changes in NPC requires further investigation.

Adult↗
PubMed1986

Diagnostic utility of lymphocyte subset analysis in AIDS case finding.

Abnormalities of lymphocyte subsets, especially low absolute number of helper T cells, are characteristically present in acquired immune deficiency syndrome (AIDS). Similar abnormalities can be found in patients with persistent generalized lymphadenopathy (PGL) or AIDS-related complex (ARC) and, to a lesser degree, in asymptomatic people who have been exposed to human T-lymphotropic virus type III (HTLV-III). Nevertheless, there appears to be a widespread perception that lymphocyte subset analysis may be useful in AIDS case finding within high-risk groups. We evaluated the diagnostic utility of absolute number of helper T cells and ratio of helper to suppressor T cells in 33 patients with AIDS, 43 patients with PGL who had been referred for lymph node biopsy, 90 patients with PGL and 195 male homosexual controls. At conventional cutoff levels the tests did not appear to revise the probability of AIDS upward to any clinically significant degree when the pretest probability of AIDS was low. Lymphocyte subset analysis does not appear to be a cost-effective method of AIDS case finding in identified groups at risk in which the prevalence of AIDS is low.

Acquired Immunodeficiency Syndrome↗
PubMed1993

The effect of light, moderate and severe bicycle exercise on lymphocyte subsets, natural and lymphokine activated killer cells, lymphocyte proliferative response and interleukin 2 production.

The purpose of the present study was to evaluate the effect of acute bicycle exercise at different exercise intensities on the immune system. Six healthy volunteers exercised on a bicycle ergometer for 1 h at 25%, 50% and 75% of VO2max with an interval of 2 to 3 weeks. Blood samples were collected in the basal state, at the end of exercise and 2 h later. The absolute concentrations of all lymphocyte subsets increased during and fell after exercise at 50% and 75% of VO2max, but did not change significantly at 25% of VO2max. However, at all exercise levels, the percentage of CD3+ blood mononuclear cells decreased due to a decline in the fraction of CD4+ cells. This decline was most pronounced at 75% of VO2max. The fraction of NK cells expressing either the CD16 or the CD56 marker increased during exercise and declined to prevalues 2 h later, however the changes were most pronounced at 75% of VO2max. The natural killer (NK) cell and lymphokine activated killer (LAK) cell activities (lysis per fixed number of mononuclear cells) were increased during all exercise intensities, but were only suppressed below basal levels after exercise at 75% of VO2max. Indomethacin in vitro abolished the post-exercise suppression of NK cell activity and the proportion of CD14+ monocytes increased 2 h after exercise only at 75% of VO2max. These findings indicate that after exercise NK cell function is inhibited by prostaglandins released by monocytes. During exercise at 50% and 75% of VO2max the proliferative response of blood mononuclear cells (BMNC) following stimulation with phytohaemagglutinin A (PHA) decreased, whereas that following stimulation with interleukin-2 (IL-2) was enhanced. The IL-2 production by BMNC in vitro was markedly decreased during and after exercise at 75% of VO2max and this inhibition could be abolished by indomethacin in vitro. In conclusion, the response of the immune system to exercise depends on exercise intensity. In essence, the response is enhanced during exercise, however, after heavy exercise it is suppressed due to an increased level of prostaglandins produced by the elevated number of monocytes.

Adult↗
PubMed2006

Analysis of diurnal variation of lymphocyte subsets in healthy subjects in the Caribbean, and its implication in HIV monitoring and treatment.

Absolute values of the lymphocyte subsets are known to be influenced by various biological factors. We set out to determine if diurnal variations in lymphocyte subsets occur in our population. A prospective study was done on 25 randomly chosen healthy subjects. Persons were enrolled for the early morning to mid afternoon study i.e. 8:30, 12:30 and 15:30. In a second study, samples were collected at hourly intervals from 08:30 to 12:30. The EDTA samples were analyzed for lymphocyte subsets by flowcytomery. In the first study, the results have shown that there was a progressive increase in CD4 cell count throughout the day, while CD8 and CD19 cell counts increased between 08:30am and mid-day and then there was no further change between midday and mid afternoon. CD56 was uniform throughout the whole day. As most clinics and venesections take place in the morning, the aim of the second part of the study was to focus on the nature of the changes observed in the morning to midday phase. The results have shown that there were no significant changes in the lymphocyte subset counts before 11:30, thereafter there was a progressive increase in all of the lymphocyte subsets between 11:30 and 12:30 except for the CD56 cell count. This study has shown that diurnal rhythms influence the lymphocyte subsets in a normal population. This may have major implications in the use of CD4 subset analysis in the management of HIV infected persons as an indicator for initiation of treatment. In our setting. pending the results of diurnal variation studies on PLWHA, we have set the latest blood collection time at 11:30 am.

Antigens, CD↗
PubMedDate not supplied

Abnormalities of peripheral blood T lymphocyte subsets in polymyalgia rheumatica.

The T lymphocyte subsets were studied by monoclonal antibodies in the blood of eight patients with polymyalgia rheumatica. The percentages of circulating T cells (OKT3+) were lower when compared with normal controls (p less than 0.02), as were the proportions of suppressor/cytotoxic (OKT8+) T cells (p less than 0.001), whereas the proportions of helper/inducer (OKT4+) T lymphocytes were not changed. Consequently, the OKT4:OKT8 ratio was higher in patients with polymyalgia rheumatica than in controls (p less than 0.001). On the contrary, circulating B cells were not altered. The decrease in peripheral blood OKT3+ and OKT8+ lymphocyte subsets suggests that there is an impairment of cell-mediated immunity in polymyalgia rheumatica.

Aged↗
PubMed1988

Lymphocyte subsets in patients with brain tumors.

T lymphocyte subsets of peripheral blood were studied in preoperative patients with various types of intracranial neoplasms. The subsets were analysed using monoclonal antibodies against lymphocyte membrane markers, and flow cytometry was used to quantitate percent positive cells with the antibodies. Twenty-seven patients were selected for this study, including twelve patients with malignant primary intrinsic tumors histologically consistent with a diagnosis of malignant astrocytoma or glioblastoma multiforme, and fifteen patients with extrinsic tumors diagnosed as meningioma, pituitary adenoma, craniopharygnioma and neurinoma. Twenty-five age and sex-matched individuals without evidence of either local or systemic disorders served as control subjects. The results revealed that the OKT4/8 cell ratio was 1.4 +/- 0.4 in the malignant group, 1.8 +/- 0.4 in the benign group and 2.1 +/- 0.8 in the control group. The ratio was significantly lower in the malignant group than in the control group (p less than 0.05). Leu-11+ cells were found to be 9.7 +/- 4.7 in the malignant group, 9.0 +/- 3.4 in the benign and 7.8 +/- 2.7 in the control group. These results showed that Leu-11+ cells in the patients with malignant tumors were significantly increased in comparison with the control group (p less than 0.05). The alteration of the lymphocyte subsets is considered to be an effect of neurohormones in balance through the transmission function of the brain-endocrine axis to lymphocytes in immunomodulation.

Antibodies, Monoclonal↗
PubMed1998

[Lymphocyte subsets in blood donors].

OBJECTIVES: To establish the normal values among the different lymphocyte subsets in peripheral blood, measured by surface antigen expression, in healthy population. To observe if there are potential differences in respect of age, gender or sample origin. To compare the absolute lymphocyte number obtained with flow cytometry and with a cellular counter. MATERIAL AND METHODS: Longitudinal and prospective study performed with 100 samples of blood donors. Direct immunofluorescence with triple color staining was made on whole blood, red blood cells were then lysed and samples were analysed with a flow cytometer. The lymphocyte subsets studied were the T lymphocytes and their subsets (CD4 and CD8 lymphocytes), the B lymphocytes and Natural Killer cells population. The absolute lymphocyte count was performed with an automatic cellular counter. Donor data such as age, gender and origin were recorded and were statistically analysed. RESULTS: Normal ranges from the studied parameters are similar to other series. Comparison with gender, origin or age groups gave no significant difference, although there seems to be a tendency to decrease with the ageing of the population. Total absolute lymphocyte number did not differ between the results from the cellular counter or from the flow cytometer. CONCLUSIONS: We found no differences in the absolute lymphocyte number nor in the lymphocyte subsets studied (T lymphocytes, CD4 lymphocytes, CD8 lymphocytes, B lymphocytes and NK cells) with respect to gender, age or sample source. We have established reference ranges for our laboratory. We have not found significant differences in the absolute lymphocyte number measured with cell counter or with flow cytometer.

Adolescent↗
PubMed1997

Lymphocyte subsets in experimental hemochromatosis.

The distribution of lymphocyte subsets in blood, thymus, spleen, mesenteric lymph nodes, Peyer's patches and bone marrow was evaluated in an experimental model of secondary hemochromatosis in rats. The values of CD2, CD4, CD8, B and NK cells in these different lymphoid compartments did not differ between the control group and the experimental group. These results suggest that the abnormalities of lymphocyte subsets previously reported in patients with secondary hemochromatosis may be due to factors other than iron overload.

Animals↗
PubMedDate not supplied

Reference ranges of lymphocyte subsets of healthy adults in Turkey.

UNLABELLED: The aim of this study was to determine the reference ranges of lymphocyte subsets in serologically HIV-negative healthy adults in Turkey. MATERIALS AND METHODS: Blood samples from 220 healthy adults, 105 female and 115 male, collected into tubes containing EDTA were investigated for lymphocyte subsets using flow cytometry. The age range was 18-80 years (44.80 +/- 16.69). RESULTS: The mean percentage and absolute values of the lymphocyte subsets were as follows: CD3: 72.70 +/- 8.44%, 1,680 +/- 528 cells/microl; CD4: 47.37 +/- 9.10%, 1,095 +/- 391 cells/microl; CD8: 28.99 +/- 5.99%, 669 +/- 239 cells/microl; CD19: 10.96 +/- 4.44%, 254 +/- 122 cells/microl and CD56: 7.03 +/- 3.26%, 161 +/- 92 cells/microl, respectively. The ratio of CD4/CD8 was 1.68 +/- 0.43. There was no statistically significant difference in the percentages and absolute values of lymphocyte subsets between the genders (p > 0.05). CONCLUSION: Immunophenotyping has been used to establish reference values of lymphocyte subsets in normal healthy adults in Turkey.

Adolescent↗
PubMedDate not supplied

Circulating lymphocyte subsets in major depression and dysthymia with typical or atypical features.

OBJECTIVE: Inconsistent results have been reported concerning circulating lymphocyte subsets in depression. To establish whether the immune alterations in depression could be related to neurovegetative symptoms, lymphocyte subsets were assessed in major depressive and dysthymic patients who exhibited either typical or atypical features (ie, the latter characterized by mood reactivity and reversed neurovegetative features). METHOD: Blood was collected from major depressive, atypical depressive, typical dysthymic, or atypical dysthymic patients and from nondepressed control subjects. Circulating lymphocyte subsets (CD3, CD4, CD8, CD19, CD16/CD56) were determined by flow cytometry. In a subset of patients, lymphocyte subsets were also determined after a 12-week course of antidepressant medication. RESULTS: Although T and B cell populations did not differ between the depressive subtypes and control subjects, circulating natural killer (NK) cells were elevated in depressive illness, and varied as a function of depressive subtype and sex. Among male patients, NK cells were elevated to a greater extent in typical than in atypical depression, and more so in major depressive than in dysthymic patients. Among female patients, circulating NK cells were lower than in male patients, and only among the typical major depressive patients did NK cells exceed those of controls. Normalization of NK cells occurred with successful pharmacotherapy. CONCLUSIONS: Depression may be associated with elevated levels of circulating NK cells. Although the neurovegetative features associated with depression, particularly altered eating, may have contributed to the elevated NK cells, depressive affect itself also contributed in this respect. However, the relative contributions of these factors varied between male and female patients.

Adult↗
PubMed1996

Peripheral blood lymphocyte subsets in idiopathic nephrotic syndrome of childhood.

Lymphocyte subsets in the peripheral blood were examined in 34 patients with nephrotic syndrome [27 with minimal change nephrotic syndrome (MCNS), 7 with significant lesions] and 10 normal children. The investigations were done at the onset of disease before administering corticosteroids (n = 34), during the subsequent remission (n = 27), and the first relapse (n = 12). The proportions of T cell subsets (CD3, CD4 and CD8) were determined by immuno-alkaline phosphatase staining; interleukin-2 receptor (IL-2 R) positive cells, natural killer (NK) cells and B cells were studied by indirect immunofluorescence. No significant differences were found in the proportion of CD3, CD4, CD8 and IL-2 R positive cells in patients with MCNS as compared to controls. An increased proportion of NK and B cells was not specific to MCNS and was detected even in those with significant renal lesions. Our findings suggest that T lymphocyte subsets in MCNS are similar to those in normal children.

Adolescent↗
PubMed1992

Lymphocyte subsets in HTLV-II-infected former blood donors: relationship to spontaneous lymphocyte proliferation.

Previous studies showed that over 70% of HTLV-seropositive blood donors from the Los Angeles area are infected with HTLV-II; further, mononuclear cells from about half of these HTLV-II+ donors exhibit spontaneous lymphocyte proliferation (SLP) during in vitro culture. To determine if HTLV-II+SLP+ donors exhibit more marked immune system changes than HTLV-II+SLP- donors, lymphocyte subsets for these two HTLV-II+ groups were compared to an uninfected control group. The percentage of lymphocytes expressing CD3 was significantly increased and the percentage expressing a CD16/56+CD3- phenotype (natural killer cells) was significantly decreased in the HTLV-II+SLP+ group (N = 34) versus the control group (N = 49). On the basis of absolute numbers, the lymphocyte number was significantly higher in the HTLV-II+SLP+ group than in the control group and reflected significant increases in the numbers of both CD4 and CD8 subsets of T cells. Analysis of proportional changes in CD4 and CD8 cell subsets revealed significant increases in the proportions of CD4 cells expressing HLA-DR, CD8 cells expressing HLA-DR, and CD8 cells expressing CD45RO for the HTLV-II+SLP+ group versus the control group. For all phenotypic parameters measured, no significant differences were noted when comparing the HTLV-II+SLP- group (N = 21) and the control group. Cell culture experiments utilizing purified CD4 cells and CD8 cells from a subset of each study group revealed that in vitro spontaneous proliferative capacity resides within both the CD4 cell and CD8 cell populations from SLP+ individuals. These findings show that changes in circulating lymphocyte subsets in HTLV-II infection are found only in association with SLP, and that the capacity to exhibit SLP characterizes both CD4 and CD8 lymphocyte subsets.

Blood Donors↗
PubMed1999

Primary and anamnestic responses of bovine bronchoalveolar and peripheral blood lymphocyte subsets to aerosolized Pasteurella haemolytica A1.

Site-specific responses of bronchoalveolar and peripheral blood lymphocyte subsets were compared during primary and anamnestic immune responses against live Pasteurella haemolytica A1 (Ph1). Eight 1-year old calves were sequentially exposed intrabronchially with aerosolized Ph1 on days 0, 14, and 21, and two calves were sham exposed. Bronchoalveolar and peripheral blood lymphocytes were analyzed before each Ph1 exposure, and on days 3 and 7 post exposure using single and two-color flow cytometry to identify CD2+, CD4+, CD8+, CD21+, CD45R+, CD25+ and gammadelta lymphocyte subsets. Significant differences (p < 0.05) in bronchoalveolar and peripheral blood lymphocyte subsets were observed before Ph1 exposure. Subsequent aerosol exposures, resulted in significant (p < 0.05) changes in bronchoalveolar lymphocyte subsets and the CD4:CD8 bronchoalveolar lymphocyte ratio, but concomitant changes were not observed in peripheral blood lymphocytes. Expression of CD2, CD4 and CD8 lymphocyte differentiation antigens was consistently lower and more heterogeneous on bronchoalveolar lymphocytes. Differential analysis of bronchoalveolar leukocytes revealed a significant increase in bronchoalveolar lymphocytes and neutrophils during anamnestic responses.

Aerosols↗