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PubMed1983

Characterization of chicken lymphocyte subsets separated by peanut agglutinin.

The reactivity of chicken lymphocytes isolated from bursa, thymus, spleen, and peripheral blood (PBL) with peanut agglutinin (PNA) has been investigated. High numbers of cells binding PNA (PNA+ cells) were found in bursa (74.5%) and thymus (85.2%), as well as in the peripheral organs (spleen and PBL, 43.8 and 70%, respectively). In the latter organs, the levels of PNA+ cells exceeded by far the values reported for mammals. Separation of PNA+ and PNA- cells from different organs was achieved by selective agglutination with the lectin or by affinity chromatography on immobilized PNA. Experiments combining the use of limiting concentrations of neuraminidase to cleave off sialic acid from the surface membrane of splenocytes that do not bind PNA (PNA- cells), followed by agglutination with the lectin, suggested that the high binding of PNA to mature lymphocytes is most likely due to a low degree of sialylation of chicken cell surface glycoconjugates. The functional properties in vitro of PNA+ and PNA- spleen cell fractions were assayed in mitogen responses (PHA, Con A, PWM), the mixed-lymphocyte reaction (MLR), and the in vitro anti-SRBC antibody response. In all three systems the responses of the PNA+ fraction were found to be significantly higher than those of the PNA- cells. Mixing experiments revealed that irradiated (1500 R) PNA- cells preferably suppress pure T-cell responses of unfractionated spleen cells, whereas PNA- cells exerted a stronger suppressive effect on responses involving B cells. Thus, in the chicken, PNA may be a valuable tool to distinguish subsets of suppressor cells with different target specificity.

Animals↗
PubMed1993

The kinetics of lymphocyte subsets and macrophages in subarachnoid space after subarachnoid hemorrhage in rats.

BACKGROUND AND PURPOSE: Although it has been suggested that humoral immunity plays a role in the pathogenesis of cerebral vasospasm after subarachnoid hemorrhage, there has been no quantitative assay for cellular immunity. We studied the kinetics of immune cells in the subarachnoid space after subarachnoid hemorrhage in the rat. METHODS: One hundred fourteen Sprague-Dawley rats were used in this study. The animals were divided into two groups and injected with either autologous blood (0.3 mL) or saline into the major cistern. They were killed at the specified time: 10 minutes or 1, 2, 3, 5, or 7 days after subarachnoid hemorrhage. For immunohistochemical analysis, the rats' whole brains were frozen, and cryostat sections were prepared. For flow cytometric analysis of immune cell presence, their whole brains underwent enzymatic digestion. RESULTS: Histopathologic study revealed pathological change of the arterial wall, and immunohistochemical study revealed the existence of macrophages and T cells in the subarachnoid space in animals with a survival time of 2 to 5 days after subarachnoid hemorrhage. A flow cytometric study revealed the peak of appearance of T cells and macrophages 2 days after subarachnoid hemorrhage. The helper-suppressor T cell ratio also reached a peak 2 days after subarachnoid hemorrhage. CONCLUSIONS: A serial response of immunoreactive cells, which resembles that of the chronic allergic reaction observed in autoimmune diseases or delayed-type hypersensitivity, exists in the subarachnoid space after subarachnoid hemorrhage. The present results suggest that the initial response in cellular immunity, which is followed by humoral immunity and eicosanoid reactions, plays a role in eliciting the development of cerebral vasospasm.

Animals↗
PubMed1992

Lymphocyte subsets in immune-mediated otitis media with effusion.

To examine the role of T-cell subsets in immune-mediated otitis media with effusion induced by keyhole limpet hemocyanin (KLH), we used immunohistochemical methods to investigate the kinetics of immunocytes of the middle ear (ME) and eustachian tube (ET) in healthy BALB/c mice. Antibodies against murine macrophages and granulocytes (anti-Mac-1), helper T cells (anti-Lyt-1), suppressor T cells (anti-Lyt-2), immunoglobulins (anti-IgG, -IgM, -IgA), secretory component (SC) and KLH were used. The ME exhibited a substantial immune response, whereas the response of the ET was minor and was associated with a secondary ME immune response. After KLH challenge, an effusion with an extensive infiltration of inflammatory cells (Mac-1, IgG+ and IgM+ cells) was observed at days 1 and 3 in the ME cavity and rapidly disappeared by day 7. Within the ME mucosa, a large number of cells was observed at days 1 and 3, peaking on day 7 when a submucosal lymphoid infiltration was detected. In the immune response of the ME mucosa, Mac-1 cells were the predominant cell type followed by helper T cells, IgG+ cells, IgA+ cells and then IgM+ cells. Suppressor T cells were rarely detected after KLH challenge. SC was present within ME epithelial cells from days 1 to 14.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1993

Accessory molecules involved in antigen-mediated cytolysis and lymphokine production by cytotoxic T lymphocyte subsets. I. Identification of functions for the T cell surface molecules Ly-6C and Thy-1.

The murine T cell surface molecules Ly-6C and Thy-1 are genetically and structurally distinct, yet they share two interesting properties: both are attached to the plasma membrane through a glycophosphatidylinositol linkage, and some mAb reactive with these molecules can activate T cells. Although mAb for Ly-6C and Thy-1 appear to mimic the function of physiologic ligands, direct evidence for the existence of these putative ligands has not been presented. In this report, we describe CTL clones that use Ly-6C and Thy-1 as accessory molecules for activation of cytolysis and the production of IFN-gamma based on inhibition of these functions with mAb. These studies were facilitated by the derivation of a nonactivating hamster IgM mAb specific for Ly-6C. CTL clones that use Ly-6C and Thy-1 as accessory molecules include a subpopulation of the previously described CD8+ alloreactive CTL that are not inhibited by mAb reactive with CD8, a CD8+ TCR-alpha/beta+ T cell clone specific for HSV glycoprotein D, and a CD4-CD8- TCR-gamma/delta+ T cell clone specific for HSV glycoprotein I. The role of Ly-6C and Thy-1 in target cell recognition is to some degree tissue-specific with respect to the APC/target cell. A mAb specific for Ly-6C appears to inhibit activation by prevention of adhesion between the effector cells and the target cells. This is the most direct evidence to date of a functional ligand for Ly-6C.

Animals↗
PubMed1990

Beta-adrenergic receptors in lymphocyte subsets after exercise. Alterations in normal individuals and patients with congestive heart failure.

Dynamic exercise increases the number of beta-adrenergic receptors in mixed lymphocytes by a mechanism that is incompletely understood. In a set of in vivo studies, we have investigated the effects of dynamic exercise on the subset distribution of circulating lymphocytes and on the number of beta-adrenergic receptors in each of these subsets in two groups of patients. In healthy subjects, exercise increased plasma norepinephrine and epinephrine and caused lymphocytosis. Whereas the number of Thelper cells increased only modestly, the number of Tsuppressor/cytotoxic and natural killer cells more than tripled. The number of beta-adrenergic receptors varied among subsets but was not significantly altered by dynamic exercise in any subset except natural killer cells (35% increase, p = 0.0302). In a group of patients with congestive heart failure, dynamic exercise increased plasma norepinephrine but did not alter plasma epinephrine and did not cause significant lymphocytosis. We did not detect any significant alterations of circulating leukocyte subsets or beta-adrenergic receptors in any of these subsets after exercise. A combined analysis of healthy patients and heart failure patients revealed a significant correlation between increases in plasma epinephrine and increases in circulating lymphocytes. We conclude that the exercise-induced increase in beta-adrenergic receptors of mixed lymphocytes is predominantly caused by a redistribution of circulating cell subsets that differ in their beta-adrenergic receptor number. This appears to be mediated by epinephrine rather than norepinephrine.

Adult↗
PubMed1984

The effect of cyclophosphamide on T lymphocytes and T lymphocyte subsets in patients with chronic progressive multiple sclerosis.

The lymphocytes in peripheral blood and cerebrospinal fluid of patients with chronic progressive multiple sclerosis (MS) were characterized with monoclonal antibodies to surface antigens of T cells, helper/inducer T cells and suppressor/cytotoxic T cells. The influence of cyclophosphamide treatment on these immune parameters was investigated. Compared to healthy persons, the mononuclear cell fraction of the peripheral blood of patients with chronic progressive MS consisted of normal %s of T cells and helper/inducer T cells, but decreased %s of suppressor/cytotoxic T lymphocytes. Intensive as well as chronic treatment of MS patients with cyclophosphamide resulted in a decline in the %s of T cells and helper/inducer T cells, whereas the %s of suppressor/cytotoxic T cells returned to normal. In cerebrospinal fluid, cyclophosphamide also induced a relative decrease in the % of helper/inducer T cells and an increase in the % of suppressor/cytotoxic T cells compared to untreated MS patients. Intensive as well as chronic therapy with cyclophosphamide both led to a significant decrease in the absolute number of T cells and T cell subsets in the blood of the patients.

Adult↗
PubMed1983

In situ demonstration of T lymphocyte subsets in granulomatous inflammation: leprosy, rhinoscleroma and sarcoidosis.

T lymphocyte subpopulations in frozen tissue sections of four granulomatous conditions (five patients with tuberculoid leprosy, five with lepromatous leprosy, seven with sarcoidosis and four with rhinoscleroma) were studied using monoclonal antibodies and a modified immunoperoxidase technique. Two immunohistological patterns were observed. In tuberculoid leprosy and sarcoidosis, lymphocytes expressing the helper/inducer phenotype were present within the aggregates of mononuclear phagocytes (epithelioid cells); however, cells with the suppressor/cytotoxic phenotype were predominantly in the lymphocytic mantle surrounding each granuloma. In lepromatous leprosy and rhinoscleroma the helper/inducer T cells and suppressor/cytotoxic T cells were both diffusely distributed among the mononuclear phagocytes (histiocytes) without any discernible mantle. The segregation of the helper/inducer and suppressor/cytotoxic phenotypic subsets was associated with an epithelioid cell differentiation of mononuclear phagocytic cells, bacterial elimination and a delayed type hypersensitivity response. The intimate admixture of helper/inducer and suppressor/cytotoxic subsets was associated with undifferentiated mononuclear phagocytes, bacterial proliferation and the absence of a delayed type hypersensitivity response. Thus the different distributions of T cell subpopulations in granulomas may be associated with differences in the host's immune response in several forms of granulomatous reactions.

Granuloma↗
PubMed1983

Characterization of human umbilical cord blood lymphocyte subsets fractionated on immobilized peanut agglutinin.

Human cord blood mononuclear cells from single donors were separated on minicolumns of peanut agglutinin (PNA) coupled to Sepharose beads to yield two fractions: unbound cells (PNA-, 78%) that were eluted with phosphate buffered saline, and bound cells (PNA+, 22%) eluted with 0.2 M D-galactose. The total yield was 86% and the cells were fully viable. There was no enrichment for macrophages or for surface immunoglobulin positive (B) cells in either the PNA+ or PNA- subset. Only 26% of the PNA+ lymphocytes formed rosettes with sheep red blood cells, in contrast to 53% of the PNA- lymphocytes. The response of the PNA+ cells to mitogens and allogeneic stimulation was considerably lower than that of the PNA- cells, while that of the latter was higher than the response of the unseparated cells. The average ratios of response of the PNA+ to PNA- cells were 0.25 for PHA, 0.20 for concanavalin A, 0.15 for pokeweed mitogen, and 0.15 In the mixed lymphocyte reaction. when tested with monoclonal antibodies to lymphocyte surface markers, it was found that the PNA+ fraction was depleted of mature T cells and enriched in Ia positive cells. Our data show that the low reactivity of human cord blood mononuclear cells may be ascribed to the presence of a subpopulation of lymphocytes which are immunologically immature. They also provide further evidence that in humans the PNA receptor is a marker for immature T or B lymphocytes.

Cell Separation↗
PubMed1984

Lymphocyte subsets in the white pulp of human spleen in normal and diseased cases.

The distribution of T and B lymphocytes and their subsets in the white pulp of human spleens extirpated from patients with cancer of the digestive tract and those with portal hypertension was examined with the appliance of monoclonal antibodies. T lymphocytes were distributed in the core of lymphatic sheath protuberance (Matsumoto), while the mantle zone of lymphatic nodule and germinal center were solely occupied by B lymphocytes. On the other hand, both T and B lymphocytes were found in the cortical zone of lymphatic sheath protuberance and outer and inner layer of lymphatic nodule. The ratio of both cells differed from one case to the other. The majority of T lymphocytes in the core of lymphatic sheath protuberance and those localized in the light region of germinal center were helper T cells. Although the lymphatic sheaths in cases with portal hypertension were narrower than those in the controls, there was no difference in the distribution density of T cells between the two groups. Hardly any B lymphocytes were found in the cortical zone of lymphatic sheath protuberance, while the mixture of T lymphocytes tended to become prominent in the outer and inner layer of lymphatic nodule in those cases.

B-Lymphocytes↗
PubMed1994

T lymphocyte subset abnormalities in peripheral blood from patients with the Guillain-Barré syndrome.

T lymphocytes are probably of pathogenic importance in many autoimmune diseases. Recently, deviations of circulating T-helper (CD4+) subpopulations have been noticed. Blood samples from 12 patients with Guillain-Barré syndrome (GBS) were studied with flow cytometry during their disease course to define circulating T cell populations. The proportion of T-helper cells (CD4+) was decreased (mean value 41 +/- 15%, P = 0.01) and the proportion of T cytotoxic/suppressor cells (CD8+) was increased (35 +/- 18%, P = 0.0006) as compared to the control group of healthy blood donors (47 +/- 8% and 26 +/- 7% respectively). The CD4+ population is divided into the helper/inducer (CD4+CD29+) and suppressor/inducer (CD4+CD45RA+) subsets, which normally are equally distributed (mean values in our control group were 45 +/- 15% and 44 +/- 15%, respectively). In patients with GBS, the helper/inducer (CD4+CD29+) subset was increased (54 +/- 10%, P = 0.05) and the suppressor/inducer (CD4+CD45RA+) subset was decreased (31 +/- 9, P = 0.005) compared to the controls. The proportion of activated HLA-DR-expressing T cells was increased (7 +/- 8%, P = 0.005) as compared to controls (3 +/- 3%). The total proportions of T cells (CD2+), B cells (CD19+) and natural killer (NK) cells (CD56+) were similar in patients and controls. The CD4+ and CD8+ populations, as well as the activated HLA-DR+ T cells, normalized during the disease course. The deviations within the CD4+ population also tended to normalize, but even at follow up after 6-33 (mean 23) months, some abnormalities remained. In conclusion, we confirm previous reports of T cell activation in peripheral blood from patients with GBS. A new finding is the deviation of T helper subpopulations with an increased helper/inducer (CD4+CD29+) subset and a decreased suppressor/inducer (CD4+CD45RA+) subset, which indicates a possible autoimmune character of GBS.

Adult↗