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PubMed1988

Ribavirin treatment of murine coxsackievirus B3 myocarditis with analyses of lymphocyte subsets.

The efficacy of the synthetic nucleoside ribavirin in the therapy of experimental myocarditis caused by coxsackievirus B3 and its effects on peripheral and splenic lymphocyte subsets were investigated. Two week old male C3H/He mice were inoculated with coxsackievirus B3. Ribavirin was administered subcutaneously on days 0 to 15 (experiment I) and on days 4 to 15 (experiment II) in a dose of 200 (experiment I: Group 2, n = 10; experiment II: Group 5, n = 10) or 400 mg/kg per day (experiment I: Group 3, n = 10; experiment II: Group 6, n = 10). Control mice (experiment I: Group 1, n = 15; experiment II: Group 4, n = 14) received an injection of saline solution. In experiment I, mice treated with ribavirin survived significantly longer than did control mice (p = 0.005, Group 1 versus Group 2; p = 0.001, Group 1 versus Group 3). In experiment II, the survival rate on day 15 in ribavirin-treated mice was high compared with that in control mice (Group 4 = 14.3%, Group 5 = 20%, Group 6 = 50%). Myocardial viral titers on days 5 to 8 were significantly lower in ribavirin-treated mice of both experiments than in control mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Identification of canine T-lymphocyte subsets with monoclonal antibodies.

A panel of five murine monoclonal antibodies to canine T-lymphocytes were produced. Antibodies 4.78, 12.125 and 8.358 reacted with approximately 18%, 39% and 60% peripheral blood lymphocytes, respectively. Two color flow cytometric analysis showed that lymphocytes expressing 1.140, 4.78, 8.53 and 12.125 were subsets of lymphocytes expressing 8.358. The lymphocytes expressing 8.358 were negative for surface immunoglobulin. The subsets defined by 1.140, 4.78 or 8.53, 12.125 were mutually exclusive and together account for most cells expressing 8.358 in the peripheral blood, spleen, and lymph node. In the thymus, approximately 47% cells were positive for both 1.140/4.78 and 8.53/12.125. SDS-PAGE analysis of radiolabelled thymus cell lysates demonstrated that antibodies 1.140 and 4.78 immunoprecipitated a 32,35 kd heterodimer under reducing conditions and 12.125 immunoprecipitated a single 56 kd chain under reducing and non-reducing conditions. Antibodies 8.53/12.125 and 1.140/4.78 react with canine lymphocyte populations that occur in proportions similar to lymphocytes expressing CD4 and CD8 like molecules in several primate and non-primate species. The molecules recognized by 12.125 and 1.140/4.78 were similar in size and subunit composition to human CD4 and CD8.

Animals↗
PubMed1989

Lymphocyte subset analysis by flow cytometry. Comparison of three different staining techniques and effects of blood storage.

Flow cytometric analysis enables the researcher and clinician to enumerate lymphocyte subsets in peripheral blood mononuclear cells (PBMC). Often blood samples are collected at one site and then shipped to another site for analysis. Many options in the storage, preparation, and staining of PBMC for flow cytometric analysis exist. Preparation techniques include the conventional Ficoll-Paque (FP) density centrifugation versus the whole blood technique with red blood cells (RBC) lysed using a lysing reagent. In comparing three methods of PBMC preparation, and comparing the staining of fresh blood cells with staining of cells after storage for 24 h at 4 degrees C, analyses show how these different techniques affect the results.

Antigens, Differentiation↗
PubMed1989

Lymphocyte subsets in bronchoalveolar lavage fluid obtained from stable asthmatics, and their correlations with bronchial responsiveness.

Bronchial responsiveness to methacholine (PD20 FEV1) was assessed in 22 asthmatic subjects approximately 5 days prior to bronchoalveolar lavage (BAL). A PD20 FEV1 could not be attained in 20 matched controls with normal pulmonary function. BAL was performed in all subjects, 3 x 60 ml aliquots of buffered saline being introduced into a segment of the middle lobe and immediately aspirated into siliconized glassware at 4 degrees C. After filtration, cells were counted, and the cell pellet resuspended in medium 199. Cytospin slides were prepared and a differential cell count performed. Lymphocyte subsets were identified by labelling further cytospin preparations with specific monoclonal antibodies (Leu series) against T3, T4, T8 and B cell markers, followed by a fluorescent antibody marker. The slides were coded, and 100 lymphocytes were then randomly scanned for fluorescence on each cytospin preparation. The median total lymphocyte counts were significantly greater in the asthmatic subjects, but this increase was confined to the T cell subgroups. The mean T4/T8 ratio was similar in asthmatic (1.51) and control (1.45) subjects. Log PD20 FEV1 correlated positively with total lymphocyte counts (r = 0.42, P less than 0.05), and with total T8 counts (r = 0.60, P less than 0.05). Correlations between bronchial responsiveness and T3, T4 and B lymphocyte numbers all failed to reach significance, and there was no correlation with T4/T8 ratios. The increase in BAL lymphocyte counts in asthma appears to be due to an absolute increase in T cell subsets, especially the T8 lymphocyte subgroup, and is most marked in mild asthmatics.

Adolescent↗
PubMed1991

Lymphocyte subset distribution after radiation therapy for cancer of the uterine cervix. Possible prognostic implications and correlation with disease course.

An analysis of lymphocyte subpopulations was done in patients with cancer of the uterine cervix before and at different intervals after the commencement of radiation therapy. A common feature was a duration of T-cell and B-cell lymphopenia after therapy. The findings relating to the T-cell subsets were interesting. Although the CD4/CD8 ratio remained unchanged in Stages I/IIA for 24 months after treatment, patients with Stages IIB and III showed a lowering of the ratio immediately after treatment. Distinctive patterns of lymphocyte subset distribution were seen in a comparison between patients who were disease-free and those with recurrent disease. The CD4+ cell counts and CD4/CD8 ratio differed between the two groups, with consistent lowered values during the follow-up associated with recurrent disease. This study demonstrates the effects of radiation therapy in altering lymphocyte subset distribution, resulting in characteristic patterns which could be used as clinical and prognostic indicators.

Antibodies, Monoclonal↗
PubMed1984

T lymphocyte subsets in osteoporosis. Effect of 1-alpha hydroxyvitamin D3.

In order to explore the possible immunological abnormalities as the background for the development of postmenopausal and senile osteoporosis, T lymphocyte subsets were identified by monoclonal antibodies of the Orthoclone OKT series. The OKT4/OKT8 (helper-inducer/suppressor T cell) ratio was significantly higher in osteoporotics with spinal compression fracture than in non-osteoporotics without such fracture in similar age. Total lymphocyte and T cell counts were similar between the two groups. Young control subjects had an OKT4/OKT8 ratio indistinguishable from elderly non-osteoporotics. Administration of 0.5 microgram/day 1 alpha-hydroxyvitamin D3 for 2 months resulted in a significant fall of the OKT4/OKT8 ratio in osteoporotics. While the significance of elevated OKT4/OKT8 in osteoporotics remains unknown, the fall of this ratio in response to administration of 1 alpha-hydroxyvitamin D3 suggests vitamin D dependence of this immunological abnormality associated with osteoporosis.

Adult↗
PubMed1987

Prodromal immune manifestations in EMC-M virus induced diabetes: islet bound and circulating antibodies, and changes in lymphocyte subsets.

The thymus-dependence of the encephalomyocarditis (EMC-M) virus induced diabetes has been demonstrated in comparative studies of normal and immunodeficient mice. Since the lymphocytic infiltration in the islets of Langerhans is modest during the virus infection, we have looked for possible indications of humoral immune mechanisms. Using fluorescence microscopy the presence of immunoglobulins in the islets could be shown 3 days after EMC-M-virus inoculation, gradually disappearing about day 14. The Ig deposit is scattered throughout the islets, but the precise target of Ig's has not been detected. Circulating islet cell surface reactive antibodies were demonstrable from the fourth day until about the third week. This period coincides largely with the period in which Ig deposits were present. Virus antibodies in peripheral blood could not be detected until the fifth day after the virus inoculation, whereas virus could be isolated from the third day. Beginning from day 5, about one third of the mice developed severe hyperglycaemia with blood glucose levels up to 35 mmol/l. Lymphocyte subsets of spleen cells were measured using a fluorescence activated cell sorter. Six days after virus inoculation the mean percentage of Lyt 2-positive (suppressor/cytotoxic) cells decreased below the value for control mice (p less than 0.05), but increased significantly (p less than 0.02) 2 weeks later.

Animals↗
PubMed1987

Differential expression of H-Y antigen on lymphocyte subsets: analysis by flow cytometry.

Expression of H-Y antigen in human white blood cells was measured using flow cytometry with monoclonal antibodies. In this system, lymphocytes were stained preferentially in the male, and to a lesser extent in the female. Analysis of the lymphocyte subsets with biotinylated H-Y antibody conjugated with streptavidin-fluorescein isothiocyanate (FITC) and subset-specific antibody conjugated with phycoerythrin derivative (RD1) revealed differential expression of H-Y among the subsets of the male. In samples from eight men, 41.1% +/- 21.7% of B cells (B1) were stained, compared with 20.7% +/- 12.8% of cytotoxic-suppressor T cells (T8) and 5.4% +/- 3.0% of helper-inducer T cells (T4). In samples from seven women, 12.4% +/- 10.9% of B cells were stained, but staining of T cells was negligible.

Adolescent↗
PubMed1990

Allo-activated CD4+ and CD8+ lymphocyte subsets: new ultrastructural findings based on computer-assisted image analysis.

Primary allo-stimulation induces substantial and differentiated modifications in the development of cytoplasmic organelles of peripheral T-lymphocyte subsets. The present computer-assisted image analysis provides evidence that the CD4+ subset undergoes significant increase in organelle density after stimulation, while the CD8+ subset exhibits a post-stimulation organelle development proportional to the cell size. Each immunological subset contains two distinct patterns of cytoplasmic organization, here defined as type I (containing a few cytoplasmic organelles) and type II (with well-developed synthesis and storage organelles). On the basis of these findings cytologic parameters could be ascribed to a heterogeneous proliferation of functionally distinct T-cell clones in CD4+ and CD8+ subsets.

CD4-Positive T-Lymphocytes↗
PubMed1986

Class II antigen expression and T lymphocyte subsets in chronic inflammatory demyelinating polyneuropathy.

The inflammatory infiltrate within human sural nerve, biopsied from six patients with active chronic inflammatory demyelinating neuropathy (CIDP) was studied for T lymphocyte subsets and Class II antigen (Ia)-expressing cells. Immunohistochemical staining with mouse monoclonal antibodies, acid phosphatase staining, and electron microscopy were used to provide an alternative assessment of macrophage and other mononuclear cell numbers. In normal control nerves Class II antigen was present upon endothelial cells, very occasional mononuclear cells and sparsely within the perineurium. In CIDP nerve dense Class II antigen staining was prominent within nerve fascicles, in capillary endothelial cells and within the perineurium. T lymphocytes of suppressor and helper type were present in small numbers only. Moderate numbers of macrophage-monocytes were found in the patients within nerve fascicles but these cells accounted for only part of the dense Ia staining. Since two nerves with hypertrophic changes, Schwann cells forming 'onion bulbs', were clearly Ia positive, the dense and widespread staining in all nerves studied is best explained by Ia antigen expression upon mononuclear and some Schwann cells.

Antibodies, Monoclonal↗
PubMed1985

In situ quantification of T-lymphocyte subsets and Langerhans cells in the inflammatory infiltrate of atopic eczema.

Tissue from the acute, non-infected eruption of sixteen atopic eczema subjects was subjected to an indirect immunoperoxidase technique using monoclonal antibodies recognizing T-lymphocyte subsets, Langerhans cells and natural killer cells. Over half the cells infiltrating the dermis were T lymphocytes, including a large majority of helper T cells and relatively few suppressor T cells. Langerhans cells were present in significant proportions in the dermis and probably reflected increased antigen presentation within the affected skin. There was no evidence of increased natural killer cell presence. This study suggests that type IV hypersensitivity may be implicated in the aetiology of atopic eczema.

Adolescent↗
PubMed2006

[Regulatory effects of yiqi zishen granule on helper T-lymphocyte subsets in IgA nephropathy patients].

OBJECTIVE: To study the pathogenesis of IgA nephropathy (IgAN) and the effect of Yiqi Zishen Granule (YZG) on the helper T-lymphocyte subsets Th1/Th2 in IgAN patients. METHODS: Intracellular cytokines secreted by Th1/Th2 were measured using flow cytometry in 24 healthy subjects and 31 IgAN patients treated by YZG before and after treatment. RESULTS: Level of interferon-gamma (IFN-gamma) was lower (P < 0.05) and levels of interleukin 4 (IL-4) and interleukin 10 (IL-10) were higher (P < 0.01) in the IgAN patients than those in the healthy controls, showing imbalance of Th1/Th2 in the IgAN patients. After YZG treatment, the level of IFN-gamma increased and IL-10 level decreased (P < 0.01). CONCLUSION: Imbalance of Th1/Th2 exists in IgAN patients. YZG has a regulatory effect on imbalance of Th1/Th2 in IgAN patients.

Adult↗
PubMed1999

Changes in CD4+ T lymphocyte subsets in circulating blood and synovial fluid following filtration leukocytapheresis therapy in patients with rheumatoid arthritis.

The purpose of this study is to determine the changes in CD4+ T lymphocyte subsets in the circulating blood and synovial fluid following filtration leukocytapheresis (LCP) therapy for patients with rheumatoid arthritis (RA). A Cellsorba column packed with polyester fibers was used for the removal of circulating leukocytes. For patients with RA, filtration LCP or sham procedures were performed 3 times with 1 week intervals between procedures. T lymphocyte surface markers in the peripheral blood and synovial fluid were measured by flow cytometry. The proportions of activated CD4+ T cells (CD4+DR+, CD4+CD25+, and CD4+CD71+) and CD4+CD29+ T cells increased significantly in the peripheral blood, but the counts of these cells were significantly reduced in the synovial fluid after 2 treatment sessions in the LCP group. No significant changes were observed in the proportion of these cells in the control group. Our findings suggest that filtration LCP may cause a redistribution of activated T cells from affected joints into the circulating blood.

Arthritis, Rheumatoid↗
PubMed1991

Peptidergic innervation of the Bursa Fabricii: interrelation with T-lymphocyte subsets.

In birds, B-lymphocytes mature in a special immune organ, the Bursa Fabricii. This organ thus offers unique possibilities for the study of the microenvironment of B-lymphocyte differentiation. We previously reported tachykinin-, vasoactive intestinal peptide-, calcitonin gene-related peptide- and galanin-immunoreactive (ir) fibres in the chicken bursa. As judged from light microscopic studies, each of the peptides was found in fibres contacting B-lymphocytes. Vasoactive intestinal peptide-ir fibres contacted macrophages. Now, we demonstrate neuropeptide Y, indicating the sympathetic nervous system, in fibres associated with arteries, not entering the follicles. CD4- and CD8-positive T-lymphocytes were dispersed in bursal follicles and the connective tissue, most densely in subepithelial regions. We could not find close apposition of fibres with either T-cell subset. We conclude that the potential neuro-immune axis in the Bursa Fabricii may represent a neuro-B-cell-link with only indirect participation of T-lymphocytes. The sympathetic input may influence the bursal microenvironment primarily by regulating the blood supply.

Animals↗
PubMedDate not supplied

Lymphocyte subsets in superficial bladder cancer: correlation with biological activity of the tumours.

BACKGROUND: The investigation of relationship of tumour biological activity and the degree of tissue infiltrating lymphocytes (T4, T8, T6) and macrophages in bladder cancer was the purpose of this study. MATERIALS AND METHODS: Tumour specimens and near and distant biopsies from 26 patients suffering from superficial bladder cancer and bladder biopsies from 6 controls were studied. Monoclonal antibodies against T helper/inducer, (CD4), T suppressor/cytotoxic (CD8), Langerhans cells (CD1a) and Monocytes/Histiocytes (LeuM5) were used for the detection of lymphocyte subpopulations. Ki67 growth fractions and recurrence rate per 100 patients-months were used for the definition of the biological activity of the tumours. RESULTS: The degree of tissue infiltrating lymphocytes differed significantly between controls and both groups. Larger (but not significantly different) numbers were noticed in the non recurrence compared to the recurrence group of patients. CONCLUSIONS: We conclude that the host's immune defence mechanism against malignant cells does not seem to produce significantly different numbers of tissue infiltrating lymphocytes in tumours of low and high aggressiveness. On the other hand the degree of lymphocyte infiltration in bladder cancer is not marked compared to controls of controls of matching age.

Aged↗
PubMed1984

Altered distribution of T lymphocyte subsets in lithium-treated patients.

37 lithium-treated patients, 6 prelithium patients and 12 healthy volunteers were studied regarding relative and absolute numbers of peripheral blood lymphocytes, T lymphocytes and T lymphocyte subsets defined with monoclonal antibodies (OKT4, OKT8). Prelithium patients did not differ significantly from healthy volunteers. The relative and absolute numbers of OKT4 cells were lower in lithium-treated patients than in the controls. The relative number of OKT8 cells increased with increasing duration of lithium therapy. Abnormally high as well as low OKT4/OKT8 ratios were found in lithium-treated patients. OKT4/OKT8 ratios were negatively correlated to the duration of lithium therapy. The urinary concentration capacity was significantly lower in patients with low OKT4/OKT8 ratio than in the other lithium-treated patients. It is concluded that lithium in therapeutic doses may have immunomodulatory effects.

Adult↗
PubMedDate not supplied

Production of leukocyte migration inhibition factor (LIF) and migration stimulation factor (MStF) by CD4+ and CD8+ human lymphocytes subsets and T-cell clones.

The production of the lymphokines leukocyte migration inhibition factor (LIF) and migration stimulation factor (MStF) at the level of CD4+ and CD8+ human lymphocyte subsets was investigated. In a first series of experiments, anti-CD4 and anti-CD8 monoclonal antibodies capable of inhibiting the activation by concanavalin-A (Con-A) of the respective T-cell subset were used. It was observed that when CD8+ cell activation was blocked, LIF was always produced after Con-A activation. When CD4+ cell activation was blocked, MStF was produced in five out of nine experiments (no activity in the other four). The addition of N-acetyl-D-glucosamine to block LIF in supernatants of anti-CD8 treated cells was unable to show evidence of masked MStF activity. In a second series of experiments, T-cell clones were established from continuous growing T-lymphocyte cell lines developed from cultures of Con-A activated normal human leukocyte cultures. The phenotype of 22 clones was determined and their ability to produce LIF or MStF investigated. Four clones produced MStF after Con-A activation and all of them were CD3+, CD4-, CD8+. Three clones produced LIF after Con-A activation and all of them were CD3+, CD4+, CD8-. We conclude that LIF is produced by CD4+ cells and MStF by CD8+ cells.

Antibodies, Monoclonal↗
PubMed1986

A standard microcytotoxicity technique for quantitative analysis of lymphocyte subsets. A comparison with indirect immunofluorescence, evaluated by microscopy or flow cytometry.

A standard complement-dependent microcytotoxicity (CDC) technique was used for quantitative analysis of T-lymphocyte subsets in human peripheral blood and the results compared to those obtained by indirect immunofluorescence microscopy and flow cytometry. The monoclonal antibodies OKT3, OKT4 and OKT8 were used in the CDC method for detection of total-T cells, T-helper and T-suppressor cells respectively. The CDC technique provided reproducible results (CV, 3-7%) correlating well with both immunofluorescence techniques. This observation was valid both for healthy persons (n = 21) and for patients (n = 10) with immunological disorders. The correct antibody dilution, correction for background and the use of eosin staining are considered critical for the usefulness of this technique. The method has several advantages: it is widely used for histocompatibility testing, only simple equipment is necessary, and the amount of monoclonal antibody required per test is small.

Antibodies, Monoclonal↗