Lymphatic subpopulations and their transition in myocardial tissue and peripheral blood of patients with biopsy-proven myocarditis

To determine abnormal immune regulation in biopsy-proven active and healed myocarditis cases, lymphatic subpopulations in myocardial tissue and peripheral blood were studied. Among 53 cases examined, 19 were active myocarditis (M) and 34 were healing or healed (HM). Five cases of myocarditis were studied sequentially. The percentages of pan T-cells, B-cells, helper/inducer T-cells (Th/i), suppressor/cytotoxic T-cells (Ts/c), etc per total marker positive cells were calculated by use of monoclonal antibodies. In myocardial tissue, the percentage of Th/i was significantly lower in HM than M (p less than 0.01). The helper/suppressor ratio (OKT4/8) in peripheral blood was 2.43 +/- 0.43 (mean +/- SE) in M, 1.61 +/- 0.19 in HM and 1.34 +/- 0.12 in age-matched controls. In 5 progressive studied cases of M, there was a decrease of the helper/suppressor ratio at 1 to 6 months after the myocarditis. It was concluded that subsidence of the immune reaction in myocardium is related to the healing process of myocarditis and may suggest improved prognosis.     

Lung and blood lymphocyte subsets in asbestosis and in mixed dust pneumoconiosis

To study the local distribution of lymphocyte subsets in inorganic dust diseases, bronchoalveolar lavage (BAL) was performed in seven patients with asbestosis, and in 13 patients with mixed dust pneumoconiosis (anthracosidero-silicosis). Lymphocyte subsets in BAL and blood were determined by the monoclonal antibodies OKT3 (pan T), OKT4 (helper/inducer), OKT8 (suppressor/cytotoxic), B1 (B-cells), OKIa (HLA-DR antigens), and Leu-7 (natural killer). The BAL lymphocytes were moderately elevated to 15 +/- 8 percent (mean +/- SD) in mixed dust pneumoconiosis, and even more markedly increased to 28 +/- 21 percent in asbestosis. The OKT4/OKT8 ratio in BAL was significantly increased to 4.5 +/- 2.1 in asbestosis, and significantly reduced to 0.9 +/- 0.8 in mixed dust pneumoconiosis. In blood, the ratio of T-cell subsets remained unchanged, though total lymphocytes were decreased in asbestosis. These results suggest altered cellular immune processes in the lungs of patients with pneumoconiosis and may indicate different immunoregulatory changes depending on the nature of the inhaled dust.     

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.     

Peripheral blood T lymphocyte subsets in leprosy

Peripheral blood T lymphocyte subsets were measured by flow cytometry in 122 patients with leprosy, in 23 normal controls, and in 27 patients with systemic lupus erythematosus (SLE). Active lepromatous patients not in reaction showed a significant lymphopenia and a significant proportionate reduction in the number of OKT3-positive (pan T), OKT4-positive (helper/inducer), and OKT8-positive (suppressor/cytotoxic) cells, but no alteration in distribution as judged by percentage and no abnormality in the helper: suppressor ratio. Borderline lepromatous subjects not in reaction had a significant selective deficiency in the number of cells of the OKT4-positive subset, with a significant but secondary lymphopenia and OKT3-positive cytopenia, a pattern similar to that found in SLE patients. Patients undergoing reversal reactions had a selective deficiency in the OKT4-positive subset in both absolute numbers and as a percentage of total lymphocytes, and a secondary deficiency in the percentage of OKT3-positive cells. No abnormalities were demonstrated in patients with active erythema nodosum leprosum, lepromatous patients with long-term treatment, and untreated or treated borderline tuberculoid patients.     

Peripheral blood T lymphocyte subpopulations defined by monoclonal antibodies in rheumatoid arthritis: distribution in patients untreated and treated by oral gold therapy

The distribution of T lymphocyte subsets was determined in peripheral blood (PB) of two groups of patients with rheumatoid arthritis by using monoclonal antibodies (OKT). In untreated patients the percentage of OKT4+ cells (helper/inducer) was found to be significantly increased as compared to healthy controls. In patients receiving oral gold therapy a similar increase in OKT4+ cells was confirmed; furthermore, these patients showed a significant decrease in OKT8+ cell population (cytotoxic/suppressor) compared to untreated patients and to normal controls. A small numerical superimposition of values of OKT4+ and OKT8+ lymphocytes was observed in untreated but not in treated patients.     

T lymphocyte subsets of the infiltrating cells in the salivary gland and kidney of a patient with Sj?gren's syndrome associated with interstitial nephritis

Lymphocyte subsets in the salivary gland and kidney were examined in a 38 years-old female patient with Sj?gren's syndrome associated with interstitial nephritis by PAP immunoperoxidase method using monoclonal antibodies. Predominant cells of the infiltrating cells in both tissues were T lymphocytes and most of them were Ia+, OKT4+ cells (activated helper/inducer T lymphocytes). A small number of T lymphocytes were OKT8+ (suppressor/cytotoxic T lymphocytes). Moreover, we found the OKT8+ cells invading the salivary duct epithelial cells. There was no difference in the proportion of lymphocyte subsets of the infiltrating cells between the salivary gland and kidney. A similar pathologic mechanism of tissue damage, therefore, was suggested in both tissues.     

T lymphocyte subsets in patients with newly diagnosed Type 1 (insulin-dependent) diabetes: A prospective study

T lymphocyte subsets in peripheral blood from 11 newly diagnosed Type 1 (insulin-dependent) diabetic patients were studied prospectively at three time intervals: as soon as possible after diagnosis, 3 weeks and 5 months later. Lymphocytes were marked with monoclonal OKT antibodies and examined in a fluorescence-activated cell sorter. The percentage of T lymphocytes (OKT3) did not change significantly at the three study times. The percentage of helper/inducer T cells (OKT4) was high the first week after diagnosis, but decreased at the 5-month examination (p less than 0.05). The percentage of suppressor/cytotoxic T cells (OKT8) was low at diagnosis but increased at 3 weeks (p less than 0.02) and 5 months (p less than 0.01). The ratio OKT4/OKT8 lymphocytes was 2.28 at diagnosis, decreasing to 1.77 at 3 weeks and 1.87 at 5 months, compared with 1.46 for 16 age-matched control subjects. There was no significant change in the absolute number of lymphocytes. It is concluded that the distribution of T cell subsets was abnormal at the time of diagnosis, but changed towards normal within a few weeks, after which there was no significant change at 5 months. It is as yet unknown whether the high proportion of helper/inducer T cells and/or the low percentage of suppressor/cytotoxic T cells at diagnosis favour immune reactions involved in the pathogenesis of Type 1 diabetes.     

Immunohistochemical characterization of hepatic tissue lymphocyte subpopulations in liver disease

Liver biopsies from 27 patients were studied by the indirect immunofluorescence and immunoperoxidase technique for relative distribution of B cells and T-cell subpopulations. T-cell subsets were defined for OKT4(+) (helper/inducer) and OKT8(+) (suppressor/cytotoxic) cells by using mouse hybridoma monoclonal antibodies. Patients with chronic active hepatitis and those with primary biliary cirrhosis showed marked relative increments in suppressor/cytotoxic OKT8(+) lymphocytes within hepatic lymphocytic infiltrates. Other patients with acute hepatitis or those with only reactive changes also showed a relative increment in OKT8(+) T lymphocytes within hepatic biopsy material. These findings suggest that tissue lymphocytic subpopulation increments in suppressor/cytotoxic T-cell subpopulations are commonly associated with hepatic lymphocytic infiltrates and may be related to disorders of immune regulation or expression in some of these disorders.     

Lymphocyte subsets in patients with acute myopericarditis, arrhythmias and dilated cardiomyopathy

To investigate the role of immunoregulatory function in determining the clinical course of acute myopericarditis, lymphocyte subsets were analysed by laser flow cytometry in 20 patients with acute myopericarditis, 30 with various arrhythmias or atrio-ventricular block and 31 with dilated cardiomyopathy. During the healing stage of acute myopericarditis, patients with residual electrocardiographic or left ventricular wall motion abnormalities presented altered frequencies of lymphocyte subsets, increased B 1 and reduced OKT 8 positive cells with an elevated OKT 4/8 ratio. The abnormal pattern was not evident in patients with acute pericarditis nor in those with acute myocarditis who recovered completely without residual abnormalities. This observation suggested that an imbalance of helper/suppressor T cells could modulate the clinical course of acute myopericarditis, either by producing extensive and irreversible myocardial damage during acute illness or by inducing chronic smoulding myocardial inflammation. Patients with ventricular arrhythmias and left ventricular wall motion abnormalities also presented reduced suppressor/cytotoxic T cells, implying that they had been suffering from chronic smoulding myocarditis mediated by immunoregulatory dysfunction. However, we could not determine whether the imbalance of helper/suppressor T cells could mediate the progression from myocarditis to dilated cardiomyopathy, since no association was demonstrated between the abnormal lymphocyte subsets and mononuclear cell infiltration in endomyocardial biopsy sample from patients with dilated cardiomyopathy.     

T-cell subsets in malignant lymphomas and monoclonal gammopathies

159 patients with malignant lymphomas or monoclonal gammopathies were investigated for lymphocytes and their subsets using conventional surface markers and a panel of monoclonal antibodies. In untreated patients with Hodgkin's disease, non-Hodgkin lymphomas and multiple myeloma, (MM) a reduction of T-cells and especially of the "helper/inducer" subset (OKT4+) was found to be a common phenomenon. The major abnormalities occurred in advanced stages of disease. Patients previously treated by chemo- and/or radiotherapy had a further decrease of T-cells, whereas the loss of OKT4+ cells was more pronounced than that of the "suppressor/cytotoxic" lymphocytes (OKT8+). The alterations of lymphocyte subsets persisted even in long-term remitters. Comparing the lymphocyte subsets in MM and benign monoclonal gammopathies (BMG), patients with BMG showed a significant reduction in OKT8+ cells, whereas the OKT4+ population was within normal range, resulting in a significant elevation of the OKT4/OKT8-ratio compared to the controls and untreated multiple myeloma.     

Peripheral blood T lymphocyte subpopulations determined by monoclonal antibodies in active rheumatoid arthritis

The introduction of an automated flow cytofluorograph has facilitated the detection of T lymphocyte subsets because it enables a larger number of cells to be analyzed. Many authors have reported a decrease of cytotoxic/suppressor T lymphocytes in rheumatoid arthritis (RA), in contrast to the results of other workers. We believe that the discrepancy between the various studies is due to the fact that different methods and different criteria for patient selection were used. Our study comprised a larger number of patients which makes the results suitable for statistical inference. Disease activity is clearly defined and all drugs that could alter the results were excluded. The use of a flow cytometer enhances the reliability of T lymphocyte subset determination by monoclonal antibodies (OKT series). Our study confirms the reports, which suggested that the number of suppressor/cytotoxic T lymphocytes (OKT8+ cells/mm3) is decreased in patients with active RA, resulting in a high T helper/inducer lymphocyte/T suppressor/cytotoxic lymphocyte ratio (OKT4+:OKT8+). This immune balance represents an interesting feature of the disease, since several active antirheumatic drugs share a common immunomodulatory action, which normalizes the OKT4+:OKT8+ ratio. Finally, we found a good correlation between the OKT4+ cells and OKT8+ cells in the normal control population. This observation enabled us to isolate a subgroup of active patients with RA not responding to slow acting antirheumatic drugs and characterized by a low OKT4+:OKT8+ ratio.     

Altered balance of immunoregulatory T lymphocyte subsets in autoimmune thyroid diseases

Three monoclonal antibodies recognizing cell surface antigens of total peripheral (OKT3), helper/inducer (OKT4) and suppressor/cytotoxic (OKT8) T lymphocytes were used by an indirect immunofluorescence technique to enumerate peripheral T lymphocytes in 25 patients with Graves' disease (including 4 euthyroid Graves' patients), 16 patients with Hashimoto's thyroiditis and 22 normal controls. Total lymphocyte count and percentages of overall T and helper/inducer T cells among peripheral lymphocytes in these conditions showed no significant difference from those of the controls. Percentage of suppressor/cytotoxic T cells, however, was decreased in Graves' disease patients with or without hyperthyroidism. The ratio of helper/inducer T cells to suppressor/cytotoxic T cells was increased in Graves' disease population and slightly increased in hypothyroid Hashimoto's thyroiditis patients. The ratio correlated with the mitogenic response of peripheral mononuclear cells to phytohaemagglutinin, but not with the serum levels of thyroid hormones nor with the titres of thyroid autoantibodies. These findings are in accordance with the results of previous functional studies and indicate possible defects in suppressor T lymphocytes in autoimmune thyroid disease.     

Disturbances of T lymphocyte subsets preceding refractory anemia with excess blasts in transformation in a child

A 5-year-old girl with refractory anaemia with excess of blasts in transformation (RAEBT) terminating in acute nonlymphocytic leukemia is reported. The patient's serum had an inhibitory effect on normal hemopoiesis. T cell subsets defined by monoclonal antibodies revealed a decrease in helper/inducer T cell population (OKT4-positive cells) and inversion of the T4/T8 ratio. It is suggested that imbalance in immunoregulatory T cells, which has a crucial role in the control of hemopoiesis, may be a primary event in the pathogenesis of myelodysplastic syndrome which eventually develops into an overt leukemia.     

Subpopulations of T lymphocytes in Kenyan patients with visceral leishmaniasis

Populations of peripheral blood T lymphocytes from patients with Kenyan visceral leishmaniasis were studied using specifically defined antisera (monoclonal antibodies, Ortho-mune OKT3, OKT4, OKT6, and OKT8). The levels of total T lymphocytes and circulating thymocytes were within the same range as those of clinically normal individuals. However, the proportions of the helper/inducer T cells were lower in untreated patients than in the controls (18.9% vs. 39.7%) while the levels of suppressor/cytotoxic T cells were higher than in the controls (40.5% vs. 27.8%). After successful antileishmania treatment these levels showed a gradual return towards normal over a period of one year. It was concluded that immunosuppression observed is due to the levels of peripheral blood helper/inducer and suppressor/cytotoxic T lymphocytes.     

Analysis with OKT monoclonal antibodies of T-lymphocyte subsets present in blood and liver of patients with chronic active hepatitis

The relative distribution of T lymphocyte subsets, as defined by the monoclonal antibodies OKT, was determined by cytofluorimetric analysis in peripheral blood and in cells isolated from liver biopsies of 31 patients with chronic active hepatitis (CAH). The percentage of peripheral blood lymphocytes binding OKT8 (directed against cytotoxic/suppressor T cells) was found to be elevated in patients with HBsAg and HBeAg positive chronic active hepatitis. Patients with CAH who had seroconverted to anti-HBe, had an increased number of OKT3-positive cells in their blood, which was directed against a common T cell surface antigen, associated with a decreased number of OKT8 positive cells. Lymphocytes isolated from liver biopsies of patients with CAH presented a general increase of OKT8-positive cells associated with a decreased number of OKT4-positive (helper/inducer) T cells. It is likely that OKT8-positive cells found in liver biopsies represent cytotoxic T cells directed against either viral or liver cell determinants.     

Analysis of T cell subsets in cancer patients treated with interferon

T cell subsets were analyzed in 33 patients with advanced cancer who were treated with either of two interferon preparations: a partially purified human leukocyte interferon (HulFN-alpha (Le] and a highly purified recombinant interferon (lFLrA). Included in the lFLrA-treated group were eight patients with immunodeficiency and Kaposi's sarcoma. The OKT4+/OKT8+ ratio was used to define the balance between helper/inducer and suppressor/cytotoxic T cell subsets. With both interferon preparations, the mean OKT4+/OKT8+ ratio decreased 24 hours after the first interferon dose. Within the HulFN-alpha (Le) group, the decrease in ratio was related to an increase in OKT8+ cells; in the lFLrA group, it was accompanied by a small decrease in the proportion of OKT4+ cells that was greater than the decrease in OKT8+ cells. Patients treated with lFLrA were followed for the first three weeks of therapy. Most patients treated with lFLrA at all dose levels, ranging from 1 X 10(6) to 54 X 10(6) units per day, had a decrease in OKT4+/OKT8+ ratio on Day 1. No substantial change in the ratio was observed on Days 7, 14, and 22. Patients with immunodeficiency and Kaposi's sarcoma had responses similar to those of patients with other cancers treated with lFLrA. In conclusion, although both HulFN-alpha (Le) and lFLrA induce immediate decreases in the OKT4+/OKT8+ ratio, the T cell subset(s) primarily responsible for the decrease varies with the source of interferon.     

Cutaneous T cell lymphoma: characterization by monoclonal antibodies

Monoclonal antibodies to human T cells permit the characterization of the surface phenotype of cutaneous T cell lymphoma (CTCL). The majority of CTCL cells are reactive with OKT1 and OKT3 monoclonals, which identify peripheral T cells and mature thymocytes. The neoplastic cells also react with OKT4, which recognizes the inducer T cell subset; they are, however, unreactive with OKT5 monoclonal, which identifies cytotoxic/suppressor T cell subsets. These data are in agreement with previous functional studies demonstrating that CTCL is a neoplasm of inducer (helper) T cells.     

Dipeptidylaminopeptidase IV (DAP IV) activity in normal and malignant T-cell subsets as defined by monoclonal antibodies

The activity of dipeptidylaminopeptidase IV (DAP IV) was investigated by a cytochemical method in isolated fractions of normal peripheral blood mononuclear cells and malignant cells from 9 patients with chronic T-cell leukaemia. A positive DAP IV reaction was observed in 52% of normal T cells, a consistent negative reaction in normal B cells and monocytes. 2 distinct T-cell subsets, helper/inducer cells (T4+/T8-) and cytotoxic/suppressor cells (T4-/T8+), were negatively selected by complement-mediated cytolysis utilizing the monoclonal antibodies OKT4 and OKT8. On examination of these T-cell subsets a positive DAP IV reaction was restricted to the majority of normal T4+/T8- cells. The malignant cells from 3 patients with T-chronic lymphocytic leukaemia, expressing the cytotoxic/suppressor phenotype (T4-/T8+) lacked DAP IV activities. In contrast, almost all leukaemic cells from 3 other cases, expressing the helper/inducer phenotype (T4+/T8-), were strongly positive. Despite their T4+/T8- phenotype, the malignant cells from 3 patients with Sézary syndrome were completely DAP IV negative. These findings suggest that the DAP IV reaction may be helpful in the further characterization of normal and malignant T-cell subsets.     

Absence of effects of cyclosporine on myocardial lymphocyte subsets in Coxsackievirus B3 myocarditis in the aviremic stage

To test the therapeutic efficacy of immunosuppression with cyclosporine upon the aviremic stage of coxsackievirus B3 (CB3) myocarditis, 2-week-old BALB/c mice were inoculated with 3 x 10(2) plaque-forming units of CB3, and the effects of cyclosporine on peripheral, splenic, and myocardial lymphocyte subsets were investigated. Cyclosporine, 25 mg/kg/day, was administered subcutaneously daily on days 10-31 (experiment 1) and days 30-51 (experiment 2). Treated groups were compared with infected controls for each experiment. In experiment 1, the survival rate of the cyclosporine-treated group was low (17/25 vs. 24/25, p less than 0.05). The severity of myocardial lesions and the distribution of lymphocyte subsets in myocardium and spleen on days 15-18 did not differ between treated and control groups; on the other hand, the percentages of peripheral Thy 1.2+ (pan T) and L3T4+ (activated helper T) cells on days 15-18 were decreased in the treated group, and those of B, Lyt 1+ (helper/inducer T), and Lyt 2+ (suppressor/cytotoxic T) subsets did not differ significantly. Notably, myocardial interleukin-2 receptor (IL-2R) positive cells, through which cyclosporine is considered to act, were scarce in both groups. In experiment 2, survival rates of two groups did not differ (treated, 32/34; untreated, 34/34; p = NS). The severity of myocardial lesions and the distribution of splenic lymphocyte subsets on days 35-38 also were not different between two groups. The percentages of peripheral lymphocyte subsets (Thy 1.2+ and L3T4+) were decreased in the treated group; those of B, Lyt 1+, and Lyt 2+ subsets did not differ significantly. In experiments 1 and 2, the thymus/body weight ratio in the treated groups was smaller than in the untreated group, but the spleen/body weight ratio in the treated group did not differ from the untreated group; histologically, medullary cellular depletion was evident in the thymus, not in the spleen, of the treated groups. We conclude that cyclosporine failed to change the distribution of lymphocyte subsets in the spleen as well as in the myocardium in CB3 myocarditis although it had effects on the peripheral blood and thymus, which may account for the higher mortality in the treated groups. The absence of beneficial effects of cyclosporine upon the CB3-infected myocardium may be related to the paucity of cyclosporine-sensitive cells (IL-2R, L3T4, and Lyt 2 positive cells) in the myocardium.     

Effect of a thymic factor on T lymphocytes in B cell chronic lymphocytic leukemia: in vitro and in vivo studies

Abnormalities of T lymphocytes in B cell chronic lymphocytic leukemia (B-CLL) have been extensively documented by several immunologic investigations. Following recent studies pointing to the favorable effect of TP-1, a partially purified extract of calf thymus, on the T cell-mediated immunity of several diseases, including Hodgkin's disease, we have used monoclonal antibodies and the enriched T lymphocytes of 16 untreated B-CLL patients to evaluate the proportion of T cell subsets before and after the administration of TP-1. In addition, the proliferative response to phytohemagglutinin (PHA) and the helper function in a pokeweed mitogen (PWM) system were assessed. In ten cases, the effect of TP-1 was also studied in vitro by evaluating the same parameters before and after incubation of B-CLL T cells with the drug. The study demonstrated that in vivo administration of TP-1 increases significantly (P less than .001) the proportion of the defective helper/inducer T cell population (OKT4-positive cells) in B-CLL, leading to a near normal OKT4/OKT8 ratio. Furthermore, the improved phenotypic profile was accompanied by an increased proliferative response to PHA and, in particular, by a significant increase (P less than .01) of T helper capacity; this increase was, however, insufficient to enable the normalization of the serum immunoglobulin levels. The in vitro incubation of B-CLL T lymphocytes did not succeed in producing significant modifications in distribution and function.