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PubMed1986

Lymphocyte subsets and Langerhans' cells in toxic epidermal necrolysis. Report of a case.

Toxic epidermal necrolysis is a life-threatening disease, the pathogenesis of which remains largely unknown. Histologically, in addition to the characteristic epidermal alterations, there is a sparse mononuclear cell infiltrate in the dermis. The immunologic characteristics of this infiltrate are not well known. In a case of drug-induced toxic epidermal necrolysis with fatal outcome in a 48-year-old man, we demonstrated that the majority of the inflammatory cells were of helper/inducer T-lymphocyte subsets, having only a minority of cytotoxic/suppressor T-lymphocytes and rare cells with natural killer cell phenotype. The significance of these observations is discussed, with reference to the occurrence of lesions at epithelial sites bearing local networks of antigen-presenting cells (Langerhans' cells).

Animals↗
PubMed1984

The tissue architecture of synovial membranes in inflammatory and non-inflammatory joint diseases. II. The localization of mononuclear cells as detected by monoclonal antibodies directed against T-lymphocyte subsets and natural killer cells.

Tissue specimens of synovial membranes from patients with rheumatoid arthritis (RA) and non-inflammatory joint diseases were analyzed with a panel of monoclonal antibodies directed towards T-lymphocyte subsets and natural killer (NK) cells. In the RA group, mononuclear cell infiltrations in the synovium presented a distinguished pattern as compared to the non-RA group. Inflammatory synovial membranes displayed an increased level of cells recognized by the monoclonal antibodies OKT4 and OKT8, especially attributable to the broadened layer of synoviocytes and to the fibrous synovial tissue. No significant difference in the RA patients was observed with regard to the percentage of OKT4 and OKT8 positive cells in different investigated compartments of the synovium, e.g., diffuse inflamed synovial tissue, fibrous synovial tissue, and perivascular infiltrations. OKT4 and OKT8 positive staining was additionally observed on spindle-shaped cells present in the fibrous and diffuse inflamed synovium. OKT10 binding cells were located in the deeper layers of synoviocytes, in the inflamed synovial tissue, and in one case in perivascular areas, whereas HNK 1 positive cells were scattered in the fibrous synovial and perivascular cells, as well as in lymphocyte clusters of synovium in RA patients.

Antibodies, Monoclonal↗
PubMedDate not supplied

Functional lymphocyte subset assessment of the Th1/Th2 profile in patients with autoimmune thyroiditis by flowcytometric analysis of peripheral lymphocytes.

BACKGROUND: Hashimoto's thyroiditis (HT) results from a parenchymal infiltration by Th1 T cell clones that ultimately may cause tissue destruction. We analysed here whether the quantitative assessment of the cytokine profile in peripheral lymphocytes could help for the evaluation of patients with HT. METHODS: We added to a flow cytometric evaluation of lymphocyte subpopulations, an assay to identify specific Th1 and Th2 functional subsets. Whole blood diluted in RPMI was activated with PMA and ionomycin for 4h at 37 degrees C in the presence of brefeldin. Immunophenotyping of samples was performed with PerCP-Cy5.5-conjugated CD3, and APC-conjugated CD4 antibodies. After staining for surface antigens, red cells were lysed and white cells were permeabilized. Intracellular cytokines were detected with FITC-conjugated INFgamma (Th1) and PE-conjugated IL-4 (Th2) monoclonal antibodies (mabs). RESULTS: Twenty-three consecutive patients were selected based on the detection of anti TPO ab concentration equal or higher than 600 UI/L. They were compared to 17 healthy control subjects (with undetectable TPO abs). The lymphocyte count and the proportion of the different lymphocyte subsets (T cells, B cells, NK cells, T-CD4+, T-CD8+, Th1 CD3, Th2 CD3, Th1 CD4, Th2 CD4 cells) were alike when both groups were compared. A significant difference appeared when the Th1/Th2 ratio measured on CD3 T cells was considered. This ratio was significantly increased in HT patients when compared to controls (24.91+/-2.9 vs. 15.5+/-1.4 (mean +/- SEM); p <0.05). When the same analysis was performed on CD4 T cells, the Th1/Th2 ratio was again higher in HT patients, although without reaching significance (13.5+/-1.9 vs. 8.8+/-0.6; p >0.05). CONCLUSIONS: Our data indicate that the Th1 context analysed in peripheral lymphocytes is dominant in HT patients. Flowcytometry could be used as a diagnostic tool to better understand the pathogeny and the outcome of destructive autoimmune thyroiditis.

Adult↗
PubMed1985

Surface antigens, SBU-T4 and SBU-T8, of sheep T lymphocyte subsets defined by monoclonal antibodies.

Monoclonal antibodies have been used to characterize molecules found on the surfaces of T cells of sheep. SBU-T4 and STU-T8 are present on 80-85% of thymocytes, absent from B lymphocytes, and present on two-thirds and one-third of lymph node T lymphocytes, respectively. Double-labelling of T lymphocytes shows that the populations recognized are mutually exclusive. Immunoprecipitation of the SBU-T4 antigen from thymocytes and T lymphocytes reveals a molecule which migrates on SDS-PAGE as a single band of 56,000 MW under both reducing and non-reducing conditions. Under reducing conditions, the SBU-T8 molecule migrates as two bands of 33,000 and 36,000 MW when immunoprecipitated from thymocytes, but only the higher MW band is present following immunoprecipitation from peripheral lymphocytes. Under non-reducing conditions, SBU-T8 migrates as dimers and other multiples. Based on immunofluorescent, immunochemical and immunohistological data, these molecules are considered to be the structural analogues of the marker molecules used to delineate T-helper and T-cytotoxic/suppressor lymphocyte subsets of other species.

Animals↗
PubMed1981

Activation of human T lymphocyte subsets: helper and suppressor/cytotoxic T cells recognize and respond to distinct histocompatibility antigens.

This study was directed at determining the major histocompatibility complex (MHC) antigens recognized by helper (Leu-3) and suppressor-cytotoxic (Leu-2) T lymphocyte subsets in man. These 2 subsets were isolated from peripheral blood with monoclonal antibodies and challenged in vitro with various stimuli. Only Leu-3 cells proliferated in response to autologous nonrosetting cells and soluble antigens, suggesting that helper but not suppressor-cytotoxic T cells recognize autologous HLA-DR antigen. Furthermore, only Leu-3 T cells responded to allogeneic DR antigen; this was shown in reactions between 2 siblings who were HLA-identical except for a single disparity at HLA-DR caused by a crossover event. Leu-3 cells activated in primary allogeneic mixed leukocyte reactions (MLR) responded equally in secondary allogeneic MLR to the priming cells and to cells that were identical at HLA-DR but discordant at HLA-A, B, and C to the priming cell. The antigens responsible for stimulating Leu-2 cells in allogeneic MLR were not identified, although the results are compatible with a role for HLA-A and B antigens and exclude a dominant role for HLA-DR. These data indicate that the helper and suppressor-cytotoxic T cell subsets in man respond differentially to MHC antigens in a manner analogous to the murine Lyt-2- 3- and Lyt-2+ 3+ populations.

Antibodies, Monoclonal↗
PubMed1987

T-lymphocyte subsets in primary lung cancer.

T-cell subsets in the peripheral blood of 63 primary lung cancer patients (23 with adenocarcinoma, 23 with squamous cell carcinoma and 17 with small cell carcinoma) and 24 normal healthy controls were determined by indirect immunofluorescence, using the monoclonal antibody reagents OKT3, OKT4 and OKT8. Correlations between T-lymphocyte subset values and stages or cell types of disease were sought. Total lymphocytes in the patient group were decreased. However, no significant difference from controls was seen in the percentage of OKT3-positive cells (Pan T-cells) in the cancer patients. The percentage of OKT8-positive cells (cytotoxic/suppressor) was increased in the early stage of disease whereas the percentage of OKT4 positive cells (inducer/helper) remained at the control level throughout all stages. The ratio of OKT4-positive to OKT8-positive T-cells (OKT4/OKT8), reflecting the balance of immunoregulatory T-cells, was, therefore, significantly decreased in patients with stage I-II lung cancer (P less than 0.05), especially in squamous cell lung cancer (P less than 0.05), whereas in stages III or IV, this T4/T8 ratio returned to the control level. In small cell carcinoma, the T4/T8 ratio was significantly decreased in stage III (P less than 0.01) and returned to the control level in stage IV.

Adult↗
PubMed1986

Methylprednisolone infusion therapy in rheumatoid arthritis patients. The effect on synovial fluid lymphocyte subsets and inflammatory indices.

Paired samples of synovial fluid (SF) and blood were obtained prior to and at 4 and 24 hours following high-dose methylprednisolone infusion therapy in a group of patients with refractory rheumatoid arthritis. After therapy there was a significant decrease in numbers of polymorphonuclear leukocytes, lymphocytes, immune complexes, and C-reactive protein in the SF. Measurement of lymphocyte subsets, using monoclonal antibodies, revealed that at 4 hours postinfusion, there was a disproportionate decrease in the percentage of SF lymphocytes expressing class II antigens (HLA-DR or Ia-like). These data suggest that glucocorticoids induce rapid changes in SF indices of disease activity and may directly influence T cell activation within the rheumatoid joint.

Aged↗
PubMed1982

The relationship between T lymphocyte subsets and Ia-like antigen positive nonlymphoid cells in early stages of cutaneous T cell lymphoma.

Immunofluorescence studies were carried out in cutaneous T cell lymphoma (mycosis fungoides) in order to analyse the microanatomical relationship of the different T lymphocyte subsets (inducer and suppressor/cytotoxic cell populations) to large nonlymphoid Langerhans-type and so-called "indeterminate" or interdigitating cells. The conventional and mouse (monoclonal) antibodies were used in various combinations using fluorescein and rhodamine labeled second layers. In 5 of the 7 cases studied the dermal infiltrate consisted of numerous T (HuTLA+) lymphocytes, 80-90% of which expressed the inducer phenotype (HuTLA+,OKT4+). Most of these cells formed close contact with large cells exhibiting large amounts of Ia-like antigens. These cells corresponded to the interdigitating and indeterminate cells in the sections. By contrast, only small numbers (10-20%) of T cells of suppressor/cytotoxic type (HuTLA+,OKT8+) were seen. These did not show a close affinity to the Ia-like antigen positive nonlymphoid component but appeared to have a predilection for the epidermis. Epidermal Langerhans cells, also strongly Ia-like antigen positive, were further defined by 2 monoclonal antibodies reacting with a cortical thymocyte antigen HTA-1. Although Langerhans cells are probably related to the Ia-like antigen positive dermal cells only a few of the abundant latter population were HTA-1+. In the remaining 2 cases, larger populations of OKT8+ (suppressor/cytotoxic) cells were seen and could be heralding a particularly benign course. These observations indicate a close functional relationship between the lymphoid and Ia-like antigen positive dermal cells during the pre-malignant phase of cutaneous T cell lymphoma.

Aged↗
PubMed1992

Lymphocyte and lymphocyte subset numbers in blood and in bronchoalveolar lavage and pleural fluid in various forms of human pulmonary tuberculosis at presentation and during recovery.

BACKGROUND: Lymphocytes have a central role in human defences against mycobacteria. A study was designed to assess the relation between lymphocyte responses and clinical pattern of disease, nutrition and recovery during treatment in patients with tuberculosis. METHODS: Lymphocyte numbers and subsets (on the basis of CD3, CD4, and CD8 monoclonal antibodies) were measured in peripheral blood and, where appropriate, bronchoalveolar lavage or pleural fluid of patients with different forms of pulmonary tuberculosis. Eleven had localised pulmonary tuberculosis, 18 miliary tuberculosis and seven a tuberculous pleural effusion. RESULTS: CD4 lymphocytes were found in greatly increased numbers in pleural fluid and were relatively depleted in the blood. Lymphocyte numbers in bronchoalveolar lavage fluid varied widely in localised pulmonary and miliary tuberculosis but were highest in lavage fluid from patients with miliary tuberculosis. This was due to an increase in CD8 lymphocytes, which were also increased in the blood. Lymphocyte numbers bore no relation to nutrition, symptom duration, or radiographic profusion scores. In miliary tuberculosis the time taken for the chest radiograph to clear (mean (SD) 17.6 (7.8) weeks) correlated with lymphocyte numbers in lavage fluid, especially CD8 cells (r = 0.74), but not with the patients' age or nutrition. After 8 weeks' treatment, total and CD4 lymphocyte numbers in lavage fluid showed a substantial increase. CONCLUSION: The association of CD8 cells with delayed recovery is compatible with suppression of the antimycobacterial action of macrophages. The switch to predominance of CD4 cells in lavage fluid during successful treatment supports the view that they may have a role in eliminating mycobacteria.

Adult↗
PubMed1988

Serum HIV antigen and anti-P24-antibodies in 200 HIV seropositive patients: correlation with CD4 and CD8 lymphocyte subsets.

Serum HIV (P24) antigen (Ag) measured by an antigen capture ELISA (Abbott) and anti-P24-antibodies (Abs) measured by a competitive ELISA (Abbott) and by Western Blot (Dupont de Nemours) analysis were correlated with lymphocyte subsets (CD4 and CD8) in 174 HIV seropositive patients without AIDS (non-AIDS) and 26 with AIDS. In the non-AIDS group, 27% of the patients were anti-P24-Ab negative and 21% were Ag positive while in the AIDS group these figures were 62% and 54% respectively (P less than 0.001). Overall, a significant correlation exists between the Ab-Ag profile and the CD4 cell count: the percentage of patients with anti-P24-Ab positive and Ag negative decreases from 90% for patients with more than 900 CD4 cells/microliter to 21% for patients with 100 and less CD4 cells/microliter; on the contrary, the percentage of patients with anti-P24-Ab negative and Ag positive increases from 0% over 800 CD4 cells to 53% under 100 CD4 cells/microliter. A weak correlation may also exist with the CD8 cell count. The subgroup of patients with 1,000 or more CD8 cells/microliter have a higher (but not significant) percentage of subjects with Ag positive and anti-P24-Ab negative than the subgroup with less than 1,000 CD8 cells/microliter. A short-term longitudinal study (mean follow-up: 1 year) was performed on 80 non-AIDS subjects: 77% (10/13) of those who had a CD4 cell decrease (greater than 30%) were initially Ag positive, while only 21% (14/67) of those without a decrease were Ag positive at the beginning (P less than 0.1). Although the relative weight and individual predictive value of each of these parameters need to be classified, they are probably the best biological markers currently available for monitoring clinical trials with experimental drugs.

Acquired Immunodeficiency Syndrome↗
PubMed2005

Interleukin-4 up-regulates T-tropic human immunodeficiency virus type 1 transcription in primary CD4+ CD38+ T-lymphocyte subset.

The capacity of human immunodeficiency virus type 1 (HIV-1) to infect resting cells and to produce progeny particles may contribute significantly to its pathogenicity in vivo. We previously reported that primary culture of resting CD4(+) CD38(+) T-lymphocyte subset had higher production rate of CXCR4-using (X4) HIV-1 than CD4(+) CD38(-) subset. Interleukin (IL)-4 highly contributed to the up-regulation of the X4 virus production in the CD38(+) subset. Here, we show evidences that IL-4 treatment of both resting CD38(+) and CD38(-) subsets allowed the adsorption, entry, and integration of X4 virus at similar rates, while the following viral transcription rate was significantly lower in the CD38(-) than CD38(+) subset. Treatment of the CD38 subsets with IL-4 or phytohemagglutinin revealed no association of X4 virus replication ability in the subsets with classic T-cell activation or proliferation. Interestingly, the activator protein (AP)-1 was significantly activated in the CD38(+) subset after IL-4 treatment, while both nuclear factor (NF)-kappaB and signal transducers and activator of transcription (STAT)-6 were activated in the IL-4-treated CD38(-) and CD38(-) subsets at similar levels. Thus, IL-4-dependent X4 HIV-1 transcription occurs efficiently in the CD38(+) but not CD38(-) subset of CD4(+) population and AP-1 could play a significant role on viral transcription, leading to the up-regulated X4 virus production in the CD38(+) subset.

ADP-ribosyl Cyclase↗
PubMed1982

Distribution of T-lymphocyte subsets in Hodgkin's disease characterized by monoclonal antibodies.

Mononuclear-cell suspensions of lymph nodes, spleen and blood from 24 patients with active Hodgkin's disease (HD) were studied for possible imbalance of T and B lymphocytes, and T-lymphocyte subsets, using monospecific anti-T antibodies and other reagents. A profile showing T-cell predominance was demonstrated in lymph nodes and blood, with total T-cells ranging from 50-70% of the cell count. As defined by monoclonal antibodies, 70-85 of the latter comprised the "inducer" subclass, the remainder being "suppressor" cells. There were no essential differences between histologically involved and uninvolved lymph nodes from HD patients, though total T-cell proportions were lower in "normal lymph node" controls. The profiles of spleens electively removed, as part of pre-treatment staging procedures, showed reduced total T-cell numbers, whether these were involved with HD or not. These differences are accounted for principally by fewer T "inducer" cells (24%, in spleen, v. 54% in involved lymph nodes and 47% in "normal" control nodes). Possible explanations for these findings are discussed. Our results demonstrate similar profiles in histologically diseased and normal tissue, rather than any clear imbalance of T-cell proportions which might explain the profound disturbances of T-cell function frequently demonstrated in vivo and in vitro.

Adult↗
PubMed1994

Blood lymphocyte subsets in rats with adjuvant arthritis.

OBJECTIVES: To determine the phenotype of peripheral blood lymphocytes during the time-course of adjuvant arthritis (AA) to detect alterations that could be involved in the pathogenesis of the arthritic process. METHODS: Phenotype analysis was performed on days 7, 14, 21, 28, 42, 56 and 70 after arthritis induction using monoclonal antibodies to CD5, CD4 and CD8 subsets, and flow cytometry. The proportion of activated lymphocytes and lymphocytes was also assessed with monoclonal antibodies to IL-2R (CD25), to Ia antigen and by polyclonal antibodies to rat Ig. RESULTS: Adjuvant arthritis produced leukocytosis with neutrophilia. Rats with AA showed a marked increase in the number of both CD4+ and CD8+ cells. The ratio CD4/CD8 decreased because the rise in CD8+ cells was more pronounced than the increase in CD4+ cells. Changes in lymphocyte counts showed two well-defined periods: the first, from day 14 to day 28, during which the inflammation of the joints reached a maximum and changes in lymphocyte subsets were more pronounced, that is, there was a threefold increase in CD8+ lymphocytes over normal counts, and the second, from day 42 to day 70, in which modified parameters improved considerably but remained different from controls. CONCLUSION: Alterations were detected in the phenotype of peripheral blood lymphocytes in AA, which provides an additional marker of disease activity.

Animals↗
PubMed1994

MHC class II and ICAM-1 expression and lymphocyte subsets in transbronchial biopsies from lung transplant recipients.

The expression of MHC class II antigens and ICAM-1 and the composition of lymphocyte infiltrates have been studied in frozen sections of transbronchial biopsies from lung transplant recipients. First, biopsies obtained from patients who showed acute rejection, OB, and normal features were compared. Second, we compared first-year biopsies from patients developing OB and patients with a good clinical outcome. HLA-DR was widely expressed on epithelia and vascular endothelium. Increased vascular HLA-DP expression was found in OB biopsies. In OB patients there was a significantly increased frequency of bronchial HLA-DP and vascular HLA-DQ expression. Expression of ICAM-1 by bronchial and bronchiolar basal cells, a phenomenon not reported previously in humans, was seen in a small number of biopsies. CD8 predominant lymphocytic infiltrates were present in all groups and were increased in OB biopsies and OB patients. Increased numbers of CD4-positive cells were found in rejection and OB when compared with normal biopsies. These findings support an immunological basis for the development of OB.

Antigens, CD↗
PubMed1994

Selective depression of T-lymphocyte subsets in gastric cancer patients: an implication of immunotherapy.

The lymphocytes and their subsets were determined in the peripheral blood of 62 gastric cancer patients and 68 controls using fluorescent conjugated monoclonal antibodies and flow cytometry (FACScan method). A significant increase in the total number of white blood cells from gastric cancer patients was noted in comparison to controls, but the percentage of lymphocytes was the same. The percentage of suppressor (cytotoxic) T cells (CD8) showed no difference in all stages of gastric cancer patients evaluated, but the percentage of helper (inducer) T cells (CD4) and the CD4/CD8 ratio decreased significantly in the advanced stages III and IV. Depression of the CD4/CD8 ratio was well correlated with tumor invasion, lymph node metastasis, and tumor size but not with sex, age, tumor location, gross type, or histologic differentiation. It appeared that the immune defect of gastric cancer patients was associated with the afferent arm (CD4) and worsened as the disease advanced. These results suggest that immunotherapy to stimulate the deficient immune system may play an important part in the multimodality treatment of patients with advanced gastric cancer.

CD4-CD8 Ratio↗
PubMed1983

Immune responses during human schistosomiasis mansoni. IX. T-lymphocyte subset analysis by monoclonal antibodies in hepatosplenic disease.

T lymphocyte immunoregulatory subsets (T4+ and T8+ cells) have been analysed in eight cases of hepatosplenic schistosomiasis mansoni, and compared to parallel analyses of seven intestinal Schistosoma mansoni-infected patients and four uninfected control subjects. The mean T4+:T8+ ratio in hepatosplenic cases was markedly less than observed in other groups (0.94 vs 1.86 vs 2.22, respectively). Proliferative responses stimulated by a soluble adult worm extract correlated directly with the observed T4:T8 ratios. Normal levels of E rosette-positive and surface immunoglobulin-bearing lymphoid cells were observed in intestinal patients, while hepatosplenic patients demonstrated normal B lymphocyte proportions but highly erratic E rosette-positive percentages. Curiously, in hepatosplenic patients the percentage of E rosette-positive cells often did not correlate with the proportion of T3+ cells.

Antibodies, Monoclonal↗
PubMed1987

Major histocompatibility antigens and lymphocyte subsets during experimental allergic neuritis in the Lewis rat.

Major histocompatibility antigens were identified in frozen sections of normal Lewis rat peripheral nerve tissue with monoclonal antibodies and an avidin-biotin-peroxidase complex system. Class I antigen is normally required for cytotoxic/suppressor T lymphocyte function and class II antigen for activation of helper T lymphocytes. In the sciatic nerves class I antigen was expressed diffusely by most endoneurial and perineurial cells but class II antigen only by a minority. In the cauda equina class I antigen was expressed by all arachnoid and some endoneurial cells, while class II antigen was expressed by a smaller proportion of arachnoid cells in the endoneurium of spinal roots and interstitial cells surrounding dorsal root ganglion neurons. The endothelium of endoneurial, perineurial and meningeal vessels uniformly expressed class I but not class II antigen. Experimental allergic neuritis was induced in Lewis rats by immunisation with bovine intradural root myelin. Early lesions consisted of multifocal infiltration of the nerve roots by cells expressing leucocyte common antigen. Surrounding endoneurial cells showed markedly increased expression of major histocompatibility antigens. In inflammatory lesions about 10% of the cells were stained with pan T cell antibodies. T lymphocyte subsets were identified with antibody W3/25 for helper cells and MRC OX-8 for cytotoxic/suppressor cells. The W3/25 positive cells were usually slightly in excess of OX-8 positive cells and their relative proportions did not alter during the disease. The presence of class I antigen on normal endothelium and its increased expression on endoneurial cells in the early phase of inflammation suggest an important role for class I restricted lymphocytes in the pathogenesis of the early stages of experimental allergic neuritis.

Animals↗