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PubMed1983

T-lymphocyte subsets during pregnancy and the menstrual cycle.

Many observations suggest that cellular immunity is depressed during gestation. To define alterations in subsets of circulating lymphocytes during pregnancy, we compared the percentages of total T cells, T helper cells, and T suppressor cells in the peripheral blood of 31 pregnant women at various stages of gestation with those in that of nonpregnant controls. Control studies consisted of five women ingesting oral contraceptives and five women menstruating spontaneously. Isolated peripheral blood mononuclear cells were reacted with fluoresceinated monoclonal antibodies that were specific for Leu-2 cells, Leu-3 cells, and Leu-4 cells and that were subsequently analyzed on a fluorescence-activated cell sorter. No variation in the percentage of Leu-2, Leu-3, or Leu-4 cells was observed during the menstrual cycle or oral contraceptive cycle. No differences were seen when T-cell subsets at various stages of gestation were compared with the corresponding T-cell subset in nonpregnant controls. The data suggest that pregnancy does not alter the percentage of T-cell subsets in the peripheral blood.

Adolescent↗

PubMed1992

Lymphocyte subsets in muscle biopsies of human T-lymphotropic virus type I carriers with polymyositis.

Recent epidemiologic studies have shown that human T-lymphotropic virus type I (HTLV-I) is associated with polymyositis. To clarify the histological features of HTLV-I positive polymyositis, we studied muscle tissue from two groups of patients with polymyositis, those seropositive for HTLV-I (5 patients) and those seronegative (5 patients). We examined the lymphocyte subsets in muscle biopsies using monoclonal antibodies. In the endomysium, a variable number of T-lymphocytes and macrophages and a smaller proportion of B-lymphocytes were found in both groups. In both HTLV-I positive and negative patients a variable number of T-lymphocytes and macrophages, with a smaller number of B-lymphocytes, were found in the perimysium. As a whole, the T-lymphocytes were predominantly of the helper/inducer phenotype in both groups. We failed to find any specific phenotype distribution among T-cells infiltrating the muscle of HTLV-I carriers with polymyositis.

Adult↗

PubMed1988

Age-related changes in human lymphocyte subsets: progressive reduction of the CD4 CD45R (suppressor inducer) population.

The purpose of this study was to investigate, by double-labeling immunofluorescence, lymphocyte subsets in the peripheral blood of healthy children, adults, and aged individuals. The absolute number of T and B lymphocytes decreases with age. The decline in T cells was attributed to a decrease in CD4 lymphocytes. We also found that the composition of the CD4 subset changes with age: in children the CD45R molecule is expressed on the majority of CD4 cells, whereas in aged subjects the absolute number of these lymphocytes is greatly reduced. The reciprocal CD4 CD29 population is not modified during the life span. Aging is also associated with the appearance of CD8 Leu 7 lymphocytes. Putative contrasuppressor cells, identified by Vicia villosa binding, represent a very small population in peripheral blood and are not subject to age-dependent variations.

Aging↗

PubMed2002

Bovine viral diarrhea virus type 1- and type 2-specific bovine T lymphocyte-subset responses following modified-live virus vaccination.

Expression of CD25 (interleukin-2 receptor alpha chain) was used to monitor antigen-specific activation of T lymphocyte subsets (CD4+, CD8+, and gamma delta T cells) from cattle immunized with modified-live virus (MLV) bovine viral diarrhea virus (BVDV) vaccines. Two groups of 15 animals each were vaccinated with one dose of either BVDV genotype 1 (BVDV-1) or BVDV-1 and BVDV genotype 2 (BVDV-1/2). Six animals negative for both BVDV antibody and BVDV virus were used as negative controls. Three animals vaccinated 7 and 5 weeks before the start of the experiment with MLV BVDV-1 vaccine served as positive controls. Blood samples were taken from the negative control group, the positive control group, and the BVDV-1/2 group 0, 21, 35, 60, and 90 days after vaccination. Blood samples were taken from the BVDV-1 group 0, 21, and 90 days after vaccination. Isolated peripheral blood lymphocytes from immunized and control animals were incubated for 5 days with and without BVDV-1 or BVDV-2. Compared with nonvaccinated animals, a significant (P <.05) increase in expression of CD25 by CD4+ (60 days), CD8+, and gammadelta T (35 to 90 days) lymphocytes from the group given BVDV-1/2 was detected following in vitro exposure to BVDV-1 or BVDV-2 after vaccination. The CD8+ and gammadelta T cells from the group vaccinated with BVDV-1 had significantly (P <.05) increased expression of CD25 compared with nonvaccinates following postvaccination exposure to in vitro BVDV-1 but not to BVDV-2. There was no significant difference between the two vaccinated groups in CD25 expression on any of the T cell subsets in response to BVDV-1 or BVDV-2 exposure. A single administration of MLV BVDV vaccine may be more effective at stimulating CD8+ and gammadelta T cell-specific immune responses to the homologous genotype than to the heterologous genotype.

Animals↗

PubMed1996

Papillon-Lefèvre syndrome. Analysis of peripheral blood lymphocyte subsets.

We have studied the peripheral blood lymphocyte populations in our 6 patients (2 female and 4 male) with a mean age of 11.19 with Papillon-Lèfevre Syndrome (PLS) using adequate monoclonal antibodies and double coloured flow cytometry. Total B, T, CD4, CD8, CD29, CD45RA, NK, HLA-DR cells were studied. Total B, T, CD4 and CD8 lymphocytes were within normal limits. We have observed an increase in the CD29 lymphocytes and NK cells and a decrease in CD45RA lymphocytes. We think that these findings are important in explaining B lymphocyte activation and in the pathogenesis of the PLS.

Alveolar Bone Loss↗

PubMed1989

Immunocytology in malignant pleural mesothelioma. Expression of tumor markers and distribution of lymphocyte subsets.

We studied the reactivity of malignant mesothelioma cells with tumor markers and the phenotypes of lymphocyte subsets in pleural effusions from 14 patients with malignant mesothelioma. For identification of cell surface antigens with monoclonal antibodies, the adhesive slide assay was used. The reaction pattern of mesothelioma cells was found to be CEA negative, Leu M1 negative, EMA positive, BMA-120 positive, My 4 positive, and BA-2 positive. The surface morphology of mesothelioma cells may be of additional help for diagnosis. By these markers, the distinction between mesotheliomas and carcinomas is facilitated. The differentiation of reactive benign mesothelial hyperplasia from malignant mesothelioma by surface marker staining is not yet possible, however. In many effusions in this study, a concomitant T-lymphocytosis was observed with a non-specific increase in the CD4/CD8 ratio, as known for other pleural diseases.

Aged↗

PubMed2001

Lymphocyte subsets in patients with idiopathic dilated cardiomyopathy.

OBJECTIVE: Although chronic myocardial inflammatory process mediated by viral and autoimmune factors has been postulated in the pathogenesis of idiopathic dilated cardiomyopathy (IDC), the role of autoimmune mechanisms still remains unclear. The aim of the present study was to investigate the rates of various T cell subsets and natural killer (NK) cells in peripheral blood in order to see whether they had a role in the immunoregulation of IDC. METHODS: The surface markers of peripheral T and B lymphocytes were detected and percentages of pan T and B cells as well as helper (CD4+) and suppressor (CD8+) T lymphocytes subsets in the peripheral blood and their ratio (CD4+/CD8+) were determined in 27 patients with IDC and in 20 healthy controls. NK cell percentage was also studied. RESULTS: There were no significant differences between IDC and control groups with respect to T and B cell percentages. The percentages of CD4+ T cell subsets were similar in both groups (48.7 +/- 8.7% vs. 43.5 +/- 9.7% respectively; p = 0.107). CD8+ T cell percentage was significantly decreased in patients with IDC than in controls (22.6 +/- 7.7% vs. 28.2 +/- 8.2%, respectively; p = 0.044). CD4+/CD8+ ratio was markedly higher in patients with IDC than controls (2.6 +/- 1.8 vs. 1.6 +/- 0.6, respectively; p = 0.006). There was no significant difference in the NK cell percentage between groups. CONCLUSION: Decreased CD8+ T cell subset is the cause of increased CD4+/CD8+ ratio, which may imply decreased self-tolerance and an immunoregulatory defect in the pathogenesis of IDC.

Adult↗

PubMed1997

Numerical and functional characteristics of lymphocyte subsets in centenarians.

The immune system in the aged is a very interesting subject for study. In this study, analysis was extended to extrathymic T cells as well as NK cells and "conventional" T cells (i.e., thymus-derived T cells) in terms of their constitution and function in both healthy and unhealthy centenarians. Middle-aged persons were used as controls. Healthy and unhealthy centenarians showed lower levels in the proportion and absolute number of lymphocytes. The major change in the constitution of lymphocyte subsets was increased levels in the proportion of NK cells (CD56+/CD57+) and extrathymic T cells (CD3+CD57+). Inversely, conventional T cells decreased in proportion and function (i.e., proliferative response to mitogen). Although NK cells increased in centenarians, NK activity by whole lymphocytes and the purified NK fraction decreased. The difference between healthy and unhealthy centenarians was small in all parameters, the only difference being a lower level of expression of CD56 antigens on CD57+ T cells in unhealthy centenarians. These results indicate that there is a major shift in lymphocyte population from conventional T cells to NK cells and extrathymic T cells with aging. Concerning the age-associated increases in CD56+ T and CD57+ T cells, these cells correspond to NK1+ T cells in mice.

Aged↗

PubMed1986

Sequential monitoring of peripheral blood lymphocyte subsets in rats.

The methodology of enumeration of T cell subsets in humans is well established and has widespread clinical application. Sequential monitoring of lymphocyte subsets in small animals requires a consistent and reliable methodology for staining and enumerating lymphocyte populations. The optimal conditions for enumerating subclasses of rat peripheral blood lymphocytes with the Spectrum III flow cytometer are described. The sources of biological variation and technical error that were considered in optimizing the techniques and the limitations in applying them are discussed.

Animals↗

PubMed2002

Characterization of lymphocyte subsets in the bronchiolar lymph nodes of BALB/c mice infected with cilia-associated respiratory bacillus.

Cilia-associated respiratory (CAR) bacillus is an unclassified, gram-negative, extracellular bacterium that causes chronic respiratory tract disease in rodents. Infected mice develop microscopic lesions characterized by a primary lymphocytic response followed by macrophage and neutrophilic infiltration. To characterize the lymphocytic subsets that respond to CAR bacillus infection, BALB/c mice were inoculated with 10(5) CAR bacillus bacteria. At seven weeks after inoculation, mice were euthanized and the tracheobronchiolar and hilar lymph nodes were collected and stained for cell surface markers to T cells (CD3, CD4, and CD8), B cells (B220, CD5), natural killer (NK) cells (pan-NK) and intracellular interleukin 10 (IL-10) and interferon-gamma (IFN-gamma). Flow cytometric analysis of lymph nodes from CAR bacillus-infected mice revealed 11% increase in frequency of B cells (R220+), 12% increase in the frequency of double-negative (CD4-CD8-CD3+) T cells, and slight increase in the B-1 subset of B cells (B220+CD5+). There was no change in the frequency of NK cells. The CAR bacillus-infected mice had an overall decrease in the frequency of T cells. Intracellular cytokine staining revealed distinct populations of T cells producing IL-10 and IFN-gamma, and IL-10 production from B cells; NK cells were not a substantial source of IFN-gamma. To our knowledge, this is the first characterization of lymphocytic responses and suggestion that B cells and double-negative T cells may be principally responsible for the lesions associated with CAR bacillus infection.

Animals↗

PubMed1991

Cytotoxic activity and phenotypic characteristics of lymphocyte subsets after therapy of cancer patients with interleukin-2.

After a 5-day period of continuous intravenous infusion of recombinant interleukin 2 (rIL-2) in seven patients with malignant melanoma or gastric or pancreatic cancer, different lymphocyte subsets were separated from patients' blood and tested ex vivo for cytotoxic activity against various tumour cell lines. Lytic activity was mediated by CD3+CD56+, CD3-CD56+, CD3-CD2+ and CD8+CD56+ lymphocytes. No cytotoxic activity could be observed within the CD3+CD56-, CD3+CD2+ or CD4+ T cell subsets. To characterize CD56+ cytotoxic cells further, the expression of other antigens on this population was analysed before and after IL-2 therapy. CD3, CD4, CD16 and CD57 antigens were weakly expressed, and the IL-2 receptor (CD25) was not detectable on these cells either before and after treatment with IL-2. In contrast, increased expression of CD2. CD8 and HLA-DR antigens occurred following therapy. The divergence of CD3 and CD8 antigen expression after IL-2 therapy was caused by an increase in CD3-CD8+ cells, detectable as a low-density CD8+ subset. This study shows that cytotoxic activity of in vivo IL-2-activated killer cells is predominantly, but not exclusively, mediated by CD3-CD56+ lymphocytes, partially coexpressing the CD8 antigen and lacking the expression of CD16 antigens.

Antigens, CD↗

PubMed1993

[Longitudinal evaluation of lymphocyte subsets and serum immunoglobulins in infants born to HIV seropositive mothers].

Early predictive factors of HIV infection in infants born to HIV infected mothers were carried out to evaluate the roles of immunological parameters for the diagnosis and prognosis of HIV infection. T-lymphocyte subsets and serum immunoglobulins were studied on cord blood in three groups of neonates: 14 infected infants, 29 sero-reverted infants and 31 control neonates. No differences were observed between the three groups. At 3 months, IgG were significantly higher in the infected infants than in sero-reverted infants. After 6 months, CD4 + cell counts, CD4/CD8 ratio were significantly lower in the infected infants and serum IgG, IgA and IgM were significantly higher in the infected group. Antigenemia p24 was detected in 78% of the infected group before 6 months. Total HIV-specific antibody persisted and progressed after 6 months. These data and viral detection appear to be complementary and useful for therapeutic strategies.

Blood Cell Count↗

PubMed1980

Characterization of lymphocyte subsets in spontaneous mouse mammary tumors and host lymphoid organs.

The subsets of small lymphocytes (surface Igm-positive or B, Thy-I-positive or T and "double-negative" or null) appearing within spontaneous mammary tumors of C3H retired breeder female mice and in various lymphoid organs of the host at different stages of tumor development were characterized directly using a radioautographic method. Tumor-bearing mice showed early transient splenomegaly and progressive atrophy of the thymus. The proportion of T small lymphocytes increased with tumor age within the tumor and in the spleen, blood and thymus; then, except in the spleen, these proportions declined again by 8 weeks of tumor growth. The incidence of B small lymphocytes showed no change in the tumor or thymus; there was an increase in the blood after 3 weeks, and a small decrease in the spleen after 7 weeks. At all the sites examined, the proportion of null small lymphocytes declined from an initial high level observed in the elderly tumor-free control mice, and then recovered to control levels after 7 weeks. The absolute numbers of T and null cells in the spleen changed in parallel to their proportions, while splenic B-cell numbers increased at 1-3 weeks. In the thymus the absolute numbers of all subsets decreased, with a late recovery of null cell numbers. Age-matched control mice showed higher proportions of null cells in the spleen, blood and thymus, and lower proportions of T cells in the blood and spleen, than did young normal mice. This spontaneous tumor appears to be characterized by an increase in T cells, rather than in null cells, as observed for rapidly growing transplanted tumors. The null cell rise in the present case takes place before the clinical appearance of tumors. In both cases, however, tumor-infiltrating B cells exhibited low levels of surface Ig, possibly related to a low level of maturation. The functional significance of these findings remains to be determined.

Animals↗

PubMed1988

The effect of burn injury and zinc nutriture on fecal endogenous zinc, tissue zinc distribution, and T-lymphocyte subset distribution using a murine model.

The interrelationship among burn injury, zinc metabolism, and circulating T-lymphocyte distribution was studied using a rat model. Sixty Sprague-Dawley male rats were fed a zinc-deficient (less than 0.5 ppm) semipurified diet and given daily subcutaneous injections of 1 mg Zn/kg body wt for 14 days. On Day 15, 24 of the rats were subjected to a full-thickness dorsal scald injury of 30% of the total body surface. Half of the burned rats were continued on the zinc supplementation (BS) while the other half were maintained on the zinc-deficient (BD) regimen by injecting physiological saline. Feces and urine were collected for 10 days postburn and subsequently analyzed for zinc content. On Day 10 postburn all the rats were sacrificed. Zinc bound to cytosol proteins in hepatic and intestinal mucosal tissue was determined by gel column chromatography procedures and T-lymphocyte subset distributions were determined by flow cytometry. No significant difference (P less than 0.05) in total endogenous zinc excretion was seen among treatment groups. A dramatic increase was seen in zinc bound to a 12,000 mol wt protein in hepatic tissue from the BS group only. The only significant (P less than 0.05) change in T-lymphocyte populations was an increase in T-suppressor cells in the BD group.

Animals↗

PubMed2004

Analysis of local T lymphocyte subsets upon stimulation with intravesical BCG: a model to study tuberculosis immunity.

Cell-mediated immune response can control tuberculosis infection. A significant role for immune cells like CD4, CD8 and gammadelta T lymphocytes have been recognized, but little is known about the kinetics of activation and accumulation of these cells in course of Tuberculosis infection in humans. This is due to both the difficult to access to human lung and the fact that most subjects are examined in different periods of infection which may condition T cell changes. To overcome these problems, we have used intravesical BCG (Bacillus Calmette-Guerin) treatment for preventing the recurrences of bladder cancer as an in vivo experimental model of human tuberculosis infection. 20 male caucasian patients with proven bladder superficial transitional cell carcinoma treated with transurethral resection followed by six weekly intravesical instillations of BCG (T0-T6) were enrolled. Changes in T lymphocyte subsets were assessed by flow cytometry in the bladder wash recovered after each BCG instillation. Our study shows that the action of BCG appears to be T cell dependent. Lymphocytes increase at any new instillation and tend towards the reduction with the suspension of the stimulus. BCG induces a massive increase in the proportion of CD4 Th1 subset followed by an increase in gammadelta T cells, while no significant variation for CD8 and NK cells is found. Our results suggest that BCG infection model represents a valid experimental tool to study the immunological events evoked in vivo by Mycobacterium tuberculosis in humans at the site of infection.

BCG Vaccine↗

PubMed1992

The changing pattern of the splenic lymphocyte subsets in tumor-bearing mice after oral treatment with OK-432.

The aim of this study was to investigate the influence of oral administration of OK-432 on the tumor growth of tumor-bearing mice. In addition, the changing pattern of the splenic lymphocyte subsets of tumor-bearing mice was evaluated by flow cytometry. OK-432 at a dose of 0.1, 1 or 10 KE was administered orally every 3 days or every other day for 30 days to subcutaneously Meth A tumor-inoculated mice. The tumor growth was significantly inhibited in the 1 KE every 3 days group, in the 1 KE every other day group and in the 10 KE every 3 days group. In the 10 KE every other day group, OK-432 inhibited the tumor growth on days 10 and 20, while the agent did not show a marked inhibitory effect on day 30. The percentages of splenic L3T4-positive cells and splenic asialo GM1-positive cells were significantly increased in the 1 KE every other day group, while the Lyt2+/Thy1.2+ ratio was decreased. On the other hand, in the 10 KE every other day group, OK-432 showed no effect on the percentages of splenic L3T4-positive cells and Lyt2+/Thy1.2+ ratio on days 20 and 30. Our results suggest that the antitumor effect of oral administration of OK-432 may be correlated with the changing pattern of L3T4-positive cells and Lyt2+/Thy1.2+ ratio.

Administration, Oral↗

PubMed2000

Identification of distinct lymphocyte subsets responding to subcellular fractions of Mycobacterium bovis bacille calmette-Guérin (BCG).

In order to investigate the ability of Mycobacterium bovis BCG vaccination to induce immune responses toward different classes of mycobacterial antigens and the cell populations involved in such responses, proliferation of distinct human lymphocyte subsets from BCG-vaccinated donors in response to different subcellular fractions of BCG was analysed and compared with that of not sensitized subjects. Proliferation of different cell subsets was evaluated by flow cytometric determination of bromodeoxyuridine incorporated into DNA of dividing cells and simultaneous identification of cell surface markers. Although a certain degree of variability was observed among different donors, after 6 days of in vitro stimulation BCG-vaccinated subjects displayed, as a mean, a stronger blastogenic response to all the classes of antigens compared with non-sensitized ones. PPD, culture filtrates and membrane antigens induced a predominant proliferation of CD4+ T cells. In contrast, preparations enriched in cytosolic antigens elicited strong proliferation of gammadelta+ T cells which, as a mean, represented 55% of the proliferating cells. Although to a lesser extent, proliferation of gammadelta+ T cells was also elicited by preparations enriched in membrane and cell wall antigens. In response to the latter preparation proliferation of CD4+ T cells and CD16+/CD3- (natural killer (NK)) cells was observed, as well. In particular, cell wall antigens were found to induce significantly higher levels of proliferation of NK cells compared with all the other classes of antigens.

Adult↗

PubMed1981

Abnormalities of T lymphocyte subsets in patients with progressive systemic sclerosis (PSS, scleroderma).

Thirty-one patients with PSS (scleroderma) were examined for evidence of abnormalities in T lymphocyte subsets. TG and TM subpopulations of peripheral blood T lymphocytes were enumerated by rosetting techniques. The helper-suppressor functions of these lymphocytes were studied in the PWM-activated in vitro assay. The absolute numbers of TG cells in patients with DS as well as those with the CREST syndrome variant of PSS were decreased in comparison with those in age-matched healthy controls (p < 0.05). The numbers of TM and T "null" cells were not significantly altered (p > 0.05). The reduction in the number of TG cells was not caused by lymphocytotoxic antibodies. The mean helper-suppressor scores for patients with PSS and DS (1.25) and those with CREST syndrome (1.36) were higher (p < 0.05) than the mean score for healthy individuals (0.80). No relationship between the immunoregulatory abnormality and the presence or titer of CICs, ANAs, hypergammaglobulinemia, or various clinical manifestations of PSS could be demonstrated in individual patients. The finding of abnormalities in T lymphocyte subpopulations and the demonstration of their altered functional expression in patients with PSS indicate that immunoregulatory mechanisms may play an important role in this connective tissue disease.

Adolescent↗