Lymphocyte subsets of the peripheral blood in Sj?gren's syndrome and rheumatoid arthritis

Lymphocyte subsets were determined in the peripheral blood from twenty-three patients with primary Sj?gren's syndrome (SS) and sixteen patients with clinically active rheumatoid arthritis (RA) by two-color flowcytometry using various monoclonal antibodies. In both diseases, T-cells (CD3+), suppressor/cytotoxic cells (CD8+) and their cytotoxic subset (CD8+CD11-) were decreased, as compared with thirty-one healthy controls. B-cells (CD 21+ and CD 3-DR+) and activated T-cells (CD 3+DR+) were increased in SS patients. Helper T-cells (CD 4+Leu8-), suppressor-inducer T-cells (CD4+Leu8+), suppressor T-cells (CD8+Leu 15+) and three natural killer (NK) cell subsets determined by both CD16 and Leu7 antibodies did not differ between controls and SS or RA, although Leu7+NK cells were significantly increased in SS patients. In addition, we found that the treatment with low-dose prednisolone decreased B-cells and suppressor-inducer T cells, and increased suppressor T-cells, cytotoxic T-cells and Leu7+NK cells. The results indicate similar changes in the proportion of lymphocyte subsets and suggest immunologically activated and deficient conditions in both diseases. Immunomodulating effects of the treatment with low-dose prednisolone on some of the lymphocyte subsets in patients with these diseases were also supported by the study.     

Immunohistological features of synovitis in ankylosing spondylitis: a comparison with rheumatoid arthritis.

The immunohistological features of the synovial membrane in ankylosing spondylitis, HLA-B27 associated oligoarthritis, and rheumatoid arthritis wer examined with particular reference to T lymphocyte subsets. T helper (CD4+) cells clearly outnumbered T suppressor/cytotoxic (CD8+) cells in rheumatoid arthritis, whereas both cell types were present in equal numbers in ankylosing spondylitis. A reduction of CD4+/CD45R+ suppressor/inducer cells relative to CD4+/UCHL1+ helper/inducer cells was apparent in all diagnostic groups. The observations were suggestive of disease specific inflammatory responses within synovial membrane.     

Expansion of CMV-specific CD8+CD45RA+CD27- T cells in B-cell chronic lymphocytic leukemia

In patients with B-cell chronic lymphocytic leukemia (B-CLL), the absolute number of T cells is increased. Although it has been suggested that these T cells might be tumor specific, concrete evidence for this hypothesis is lacking. We performed a detailed immunophenotypic analysis of the T-cell compartment in the peripheral blood of 28 patients with B-CLL (Rai 0, n = 12; Rai I-II, n = 10; Rai III-IV, n = 6) and 12 healthy age-matched controls and measured the ability of these patients to mount specific immune responses. In all Rai stages a significant increase in the absolute numbers of CD3+ cells was observed. Whereas the number of CD4+ cells was not different from controls, patients with B-CLL showed significantly increased relative and absolute numbers of CD8+ cells, which exhibited a CD45RA+CD27- cytotoxic phenotype. Analysis of specific immune responses with tetrameric cytomegalovirus (CMV)-peptide complexes showed that patients with B-CLL had significantly increased numbers of tetramer-binding CMV-specific CD8+ T cells. The rise in the total number of CD8+ cytotoxic T cells was evident only in CMV-seropositive B-CLL patients. Thus, our data suggest that in patients with B-CLL the composition of T cells is shifted toward a CD8+ cytotoxic cell type in an effort to control infections with persistent viruses such as CMV. Moreover, they offer an explanation for the high incidence of CMV reactivation in CLL patients treated with T cell-depleting agents, such as the monoclonal antibody (mAb) alemtuzumab (Campath; alpha-CD52 mAb). Furthermore, because in CMV-seronegative patients no increase in cytotoxic CD8+ T cells is found, our studies do not support the hypothesis that tumor-specific T cells account for T-cell expansion in B-CLL.     

The reappearance of 10 differentiation antigens on peripheral blood lymphocytes after allogeneic bone marrow transplantation.

Ten monoclonal antibodies and flow cytometry were applied to characterize the recovery of lymphocyte subsets in peripheral blood after allogeneic bone marrow transplantation (BMT). Ten patients were first followed for 150 days (short-term survey) and then analysed 2 years after BMT on average (long-term analysis). Eight of the 10 recipients showed increased relative and absolute numbers of CD8+ cells and reduced numbers of CD4+ cells resulting in an inverse helper/suppressor ratio. In these eight patients the CD8+ cell predominance was long-lasting and still detectable in the long-term analysis. Two patients had a normal helper/suppressor ratio throughout the study but otherwise a similar reconstitution. Despite the slow recovery of CD4+ cells, CD4+ Leu8- and CD4+ CD45RA- helper subsets were in a normal range already on day 30 and their proportions stayed higher than those of CD4+ Leu8+ and CD4+ CD45RA+ helper cells for the whole short-term survey. The number of activated suppressor cells (CD8+ HLA-DR+) increased markedly after BMT. Similarly, in eight patients high numbers of cytotoxic CD8+ CD57+ cells were found from day 50 onwards. An early and sharp rise of NK cells (CD16+, CD56+) was observed in all recipients, and seven recipients also showed an early increase in CD20+ B cells. Later on, normal or slightly elevated numbers of these cells occurred.     

A low blood lymphocyte count is associated with an expansion of activated cytotoxic lymphocytes in patients with B-cell chronic lymphocytic leukaemia

Abstract: In order to determine the relationships between CD2+ lymphocyte subpopulations and tumour mass, the immunophenotype of natural killer (NK) cells and T lymphocyte subsets was studied in 56 B-chronic lymphocytic leukaemia (B-CLL) patients and 38 healthy subjects. The patients were classified according to their blood lymphocyte count (BLC). Forty patients had BLC<30×10⁹/l (low BLC, less tumour mass) and 16 patients had BLC>30×10⁹/l (high BLC, larger tumour mass). The percentage of CD3^– CD56+ cells, as well as of CD8+, CD8+CD45RO+ and CD3+CD57+ T subsets in low BLC patients, were higher than those found in high BLC patients. Conversely, the percentages of CD3+HLA DR+, CD4+ and CD4+CD45RO+ lymphocytes were higher in high BLC patients than in low BLC patients. The CD4/CD8 ratio was decreased in low BLC patients while it was increased in high BLC patients and a significant positive correlation was found between their CD4/CD8 ratio and their BLC. We conclude that in low BLC B-CLL patients there is a decreased percentage of activated helper lymphocytes and an increased percentage of NK cells and activated cytotoxic T lymphocytes. These results suggest a role for NK cells, and helper and cytotoxic T lymphocytes in the control of tumour burden in B-CLL patients.     

Altered T-cell subset repertoire affects treatment outcome of patients with myelofibrosis

Phenotypic characterization of T cells in myelofibrosis is intriguing because of increased inflammation, markedly elevated pro-inflammatory cytokines, and altered distribution of T-cell subsets. Constitutive activation of Janus kinase-2 (JAK2) in the majority of patients with myelofibrosis contributes to the expression of the programmed cell death protein-1 (PD1) and T-cell exhaustion. We wondered whether T-cell activation affects treatment outcome of patients with myelofibrosis and sought to determine whether the JAK1/2 inhibitor ruxolitinib affects the activation of T-cell subsets. T cells from 47 myelofibrosis patients were analyzed and the percentages of either helper (CD4+) or cytotoxic (CD8+) naïve, central memory, effector memory, or effector T cells; and fractions of PD1-expressing cells in each subset were assessed. Higher numbers of T cells co-expressing CD4/PD1 and CD8/PD1 were found in myelofibrosis patients than in healthy controls (n=28), and the T cells were significantly skewed toward an effector phenotype in both CD4+ and CD8+ subsets, consistent with a shift from a quiescent to an activated state. Over the course of ruxolitinib treatment, the distribution of aberrant T-cell subsets significantly reversed towards resting cell phenotypes. CD4+ and CD8+ subsets at baseline correlated with monocyte and platelet counts, and their PD1+ fractions correlated with leukocyte counts and spleen size. Low numbers of PD1+/CD4+ and PD1+/CD8+ cells were associated with complete resolution of palpable splenomegaly and improved survival rate, suggesting that low levels of exhausted T cells confer a favorable response to ruxolitinib treatment.     

消化系恶性肿瘤病人LAK细胞和NK细胞功能与表型的变化

通过观察20例正常人和24例消化系恶性肿瘤病人外周血自然杀伤细胞(NK)和淋巴因子激活的杀伤细胞(LAK)的活性变化,以及加用重组白细胞介素2(rIL-2)刺激前后T淋巴细胞表型变化。结果发现肿瘤病人的NK细胞活性明显下降,但经rIL-2激活后LAK细胞活性得到明显提高,其溶解率接近正常水平。肿瘤病人的总T淋巴细胞(CD_(3+))和辅助/诱导T淋巴细胞(CD_(4+))水平低于正常,但抑制/杀伤淋巴细胞(CD_(8+))水平正常。辅助/诱导淋巴细胞与抑制/杀伤淋巴细胞之比为1.18,低于正常水平(1.55)。经加入rIL-2培养后,CD_(3+)和CD_(8+)淋巴细胞的比率明显升高并达正常水平。而在正常人此变化不明显,且加用rIL-2培养与不加者无显著差异。IL-2受体的表达正常人与肿瘤病人无异。结果显示胃肠道恶性肿瘤病人的免疫机制受到抑制,但能被IL-2提高至正常水平。     

Activation of T cell subsets in the peripheral blood of patients with Sj?gren's syndrome. Multicolor flow cytometric analysis.

Using 3-color flow cytometry, we determined the proportions of activated T cells (DR+), including CD4+ cells, CD8+ cells, and their subsets, in the peripheral blood of 17 patients with Sj?gren's syndrome (SS). Activated T cells were significantly increased in CD4+ cells, CD8+ cells, and T suppressor inducer (CD4+, Leu-8+), T helper (CD4+, Leu-8-), and T cytotoxic (CD8+, CD11-) subsets, but not the T suppressor/natural killer subset (CD8+, CD11+), in patients with SS as compared with the controls. Furthermore, the proportions of activated T cells in the CD4+, Leu-8- subset, the CD8+ subset, or the CD8+, CD11- subset showed a positive correlation with serum gamma globulin levels in the SS patients. Our findings suggest that a certain immunoregulatory balance between T helper and T suppressor activities is maintained in SS patients, although this balance seems to occur at highly activated levels, and that quantitative changes of some lymphocyte subsets are important factors in maintaining this balance. We discuss the possibility that CD4+, Leu-8+ cells recirculate into peripheral lymph nodes, while CD4+, Leu-8- cells migrate into inflammatory tissues such as salivary glands, since the Leu-8 antigen is reported to be a homing receptor in peripheral lymph nodes. This process might be accelerated in SS, and each T cell subset may further participate in immunologic activation in the lymph nodes or target tissues.     

Expression of NK-lineage markers on peripheral blood lymphocytes with T- helper (Leu3+/T4+) phenotype in B cell chronic lymphocytic leukemia

Heterogeneity within lymphocyte subsets expressing T-helper (T4+/Leu3+) or T-suppressor (T8+/Leu2+) markers was analyzed in 38 patients with B cell chronic lymphocytic leukemia (B-CLL) and in 11 age-matched controls. Co-expression of NK-lineage markers (M1, Leu7) on Leu2+ or Leu3+ cells was investigated by two-color immunofluorescence, and the proportion of granular lymphocytes within each subset was determined by cytochemical staining for acid phosphatase. B-CLL patients and normal controls had similar absolute numbers of cells per microL with T- suppressor phenotype. However, the proportion of Leu2+ cells co- expressing the Leu7 antigen was higher in the B-CLL patients than in the control subjects (54 +/- 3% v 27 +/- 4%, P less than .0001). The absolute number per microL of cells with T-helper phenotype was somewhat decreased in B-CLL patients compared with normal subjects (649 +/- 104 v 799 +/- 33, P less than .02), with a consequent decrease of the helper/suppressor ratio. Furthermore, co-expression of the Leu7 and, more strikingly, of the M1 markers was increased significantly on Leu3+ cells from B-CLL patients compared with normal controls (11 +/- 2% v 2 +/- 0.7%, P less than .002 for Leu7 and 40 +/- 5% v 4 +/- 1%, P less than .00001 for M1). Cytochemical studies showed that a large proportion of Leu3+ cells from B-CLL patients were granular lymphocytes, as suggested by the co-expression of natural killer (NK) cell markers. The emergence of a population of Leu3+ granular lymphocytes with NK markers, which is barely detectable in normal subjects, may provide an explanation for the impairment of T cell functions repeatedly described in B-CLL.     

Decreased CD45RA T cells in B-cell chronic lymphatic leukemia patients: correlation with disease stage.

T-cell subsets CD4, CD8 and suppressor-inducers (CD45RA) were determined in 20 patients with B-cell chronic lymphatic leukemia (B-CLL). The proportion of CD4 and CD45RA was decreased when compared with T cells from normal subjects. CD8 was markedly increased. The activity of concanavalin A-induced suppressor cells was not significantly different from that of normal controls and was negatively correlated to the percentage of CD4 of B-CLL patients. The selective loss of CD45RA cells was more prominent in patients in advanced Rai stages of the disease (III to IV) than in early stages (0 to II). Six patients of the advanced stages group suffered from autoimmune hemolytic anemia, whereas no patient in the early stages of disease showed an autoimmune phenomenon. Our results may indicate a mechanism of autoimmunity in B-CLL similar to that of patients with autoimmune diseases.     

Phenotypic and functional activation of alveolar macrophages,T lymphocytes and NK cells in patients with systemic sclerosis and primary Sj?gren's syndrome.

OBJECTIVES--Attempts to differentiate between the pathogenesis of the severe pulmonary manifestations observed in systemic sclerosis (SSc) and the mild form in primary Sjögren's syndrome (pSS) were performed by studying cell populations recovered during bronchoalveolar lavage (BAL). METHODS AND RESULTS--Two-colour flow cytometric analysis of BAL fluid lymphocytes showed a similar degree of phenotypic activation (DR+) of CD4+ and CD8+ T lymphocyte subsets and CD16+ NK cells in patients with SSc (n = 13) and pSS (n = 11) groups and healthy controls (n = 11). Alveolar macrophages expressed the CD14 antigen at significantly increased densities in patients with SSc. Alveolar macrophage activation in SSc was also suggested by increased IL-6 concentrations in neat BAL fluid and increases in macrophage production of TNF alpha and EGF in vitro. SSc patients also had increased proportions of neutrophils and eosinophils in BAL fluid. No correlations were found between any cellular subsets or cytokine levels in BAL fluid and lung status at the time of lavage in SSc or pSS patients or the subsequent course of the pulmonary function in SSc patients. CONCLUSION--It is concluded that the phenotypical activation of alveolar helper/inducer (DR+CD4+) and suppressor/cytotoxic (DR+CD8+) T lymphocytes and NK (DR+CD16+) cells is not a prerequisite for the development of lung fibrosis in SSc or bronchial hyper-responsiveness in pSS. Alveolar macrophage activation may contribute to the development of lung fibrosis in SSc.     

Morphological variants of leukemic cells in B chronic lymphocytic leukemia are associated with different T cell and NK cell abnormalities

B chronic lymphocytic leukemia (B-CLL) is a heterogeneous disease. The different morphological variants of leukemic B cells appear to define different clinical groups of patients. Several abnormalities have been found in T lymphocytes and natural killer (NK) cells from B-CLL patients. We have investigated the phenotypic and functional characteristics of purified CD2+ cells from B-CLL patients at Binet's stage A and classified according to the neoplastic B lymphocyte morphology criteria: 32 patients with typical B-CLL and 12 patients with atypical B-CLL. Forty-three age and sex matched healthy controls were also studied. In fresh purified CD2+ cells from typical B-CLL patients, percentages of CD4+, CD4+CD45RA+, CD8+CD45RA+ T lymphocytes and CD3−CD56+ (NK) cells were significantly higher than those found in atypical B-CLL patients. However, in DC2+ cells from typical B-CLL patients, percentages of CD3+, CD3+DR+, CD8+, CD4+CD45RO+, and CD3+CD56+ cells were significantly lower than those found in atypical B-CLL patients. Increased percentage of NK cells was only found in typical B-CLL patients. The proliferative response and the production of interleukin (IL)-2 and IL-4 by phytohemagglutinin (PHA) stimulated CD2+ cells were significantly higher in typical B-CLL patients than in atypical B-CLL patients. We concluded that different patterns of phenotypic and functional alterations in the T lymphocytes and NK cells of B-CLL patients are found in patients with typical or atypical B-CLL defined according to the morphology of the leukemic cells. Am. J. Hematol. 55:175–182, 1997. © 1997 Wiley-Liss, Inc.     

Expression levels of CD38 in T cells predict course of disease in male patients with B-chronic lymphocytic leukemia

CD38 expression of tumor cells has been identified as an important prognostic factor in B-cell chronic lymphocytic leukemia (B-CLL). Although CD38 is involved in effector functions of T cells, the prognostic value of CD38+ T cells has not yet been addressed in B-CLL. In the present study, CD38-expression levels in B-CLL cells and T cells from 204 patients were analyzed by flow cytometry and correlated with clinical and molecular risk parameters. CD38 expression significantly differed in the neoplastic clone from patients with low versus advanced stage, irrespective of the sex of patients. In contrast, CD38 expression was generally higher in T cells from female compared with male patients but only increased in male patients in a stage-dependent manner. In male patients, combined analysis of CD38 in T cells and B-CLL cells identified 4 subgroups with significantly different treatment-free survival. Multivariate analysis including Rai stage and molecular risk parameters of the neoplastic clone identified CD38-expression levels in T cells as an independent prognostic factor in male patients. Combined analysis of CD38 in B-CLL and T cells is superior in predicting outcome of male B-CLL patients than either parameter alone. Further studies are needed to elucidate the underlying mechanisms of the sex-specific role of CD38+ T cells in B-CLL.     

The impact of age on T cell subset profiles in patients on chronic hemodialysis awaiting renal transplantation

Age has been considered a significant risk factor in renal transplation; increasing age is associated with a higher morbidity and mortality, primarily as a consequence of infectious and cardiovascular complications. The increased incidence of potentially lethal infections in this patient population may be due to the impact of immunosuppression on a naturally declining immune system. In search for possible characteristic T cell subsets profile in aging patients, subsets of such cells were analyzed in 270 patients on chronic hemodialysis awaiting renal transplantation. Using two-color flow cytometry and murine monoclonal antibodies recognizing specific T cell subset markers, the following cell subsets were analyzed in patients < 40 yr of age (n = 229) and in patients > 55 (n = 41) years of age: total T cells (CD3); activated T cells (CD3DR); B cells (CD3-DR+); T helper/inducer (h/i) cells (CD4); T suppressor/inducer (s/i) cells (CD4+CD45+), T cytotoxic/suppressor (CD8), the activated CD4 and CD8 subsets (CD4DR and CD8DR); T cytotoxic cells (Leu2+7?); cells of the Leu-7 phenotype (Leu7+ and Leu2+7+); interleukin 2 receptor (IL-2-R) expressing CD3 and CD8 subsets (CD3IL-2?R+ and CD81L2-R+); and transferrin receptor (TR+) expressing CD3 (CD3TR+). The ratio CD4: CD8 was derived from the respective subset data. Significantly higher levels of CD3 (P < 0.005), B (P < 0.05), CD8 (P < 0.001), CD4DR (P < 0.05) and Leu2+7? (P < 0.0045) and CD8-IL-2-R+ (P < 0.005) cell subset were observed in the younger (age < 40), compared to the older (age > 55) group. In contrast, the CD4 : CD8 ratio, the s/i (CD4+CD45+), the Leu7+2- and Leu2+7+ cell subsets were found to be present at a higher level in the older group (P < 0.001, P < 0.005, P < 0.05 and P < 0.01 respectively). These data demonstrate a significant effect of aging on certain T cell subsets levels in the peripheral blood. Although this study does not provide data regarding the functional status of the immune competent cells, it highlights the actual changes in the distribution of cell subsets in the older potential organ recipients. Such changes may be responsible for the increased risk of infectious complications seen in older renal allograft recipients.     

Increased prevalence of late stage T cell activation antigen (VLA-1) in active juvenile chronic arthritis.

The presence of activated T cells as judged from the reaction with monoclonal antibodies (MoAb) against (a) a late stage T cell activation antigen (VLA-1), (b) the interleukin 2 (IL2) receptor (CD25), and (c) four different HLA class II molecules (HLA-DR, DRw52, DQ, and DP) was studied in 15 patients with active juvenile chronic arthritis (JCA), 10 patients with JCA in remission, and 11 age matched, healthy controls. In addition, the distribution of T 'helper/inducer' (CD4+), T 'suppressor/inducer' (CD4+, Leu8+), T 'suppressor/cytotoxic' (CD8+), and 'natural killer' (NK) cells (CD16+) was studied. Twenty patients and six controls were investigated for the capability to stimulate alloreactivated primed lymphocytes. The prevalence of VLA-1 positive, large cells was significantly increased to 5% (median value) in active JCA as compared with JCA in remission (2%, p less than 0.05) and controls (1%, p less than 0.05), whereas no significant difference between JCA in remission and controls was observed. Except for two patients with active JCA, less than 1% IL2 receptor bearing cells were found in patients with JCA and controls. No significant difference in the prevalence and expression of the various HLA class II antigens was observed between the groups. Similarly, no significant differences in stimulatory capability in secondary mixed lymphocyte culture (MLC) were seen. The distribution of T helper/inducer (CD4+), T suppressor/cytotoxic (CD8+), and NK cells was similar in active JCA, JCA in remission, and controls. The prevalence of T suppressor/inducer (CD4+,Leu8+) cells was higher in remission JCA (17%) than in active JCA (11%) and controls (10%). This increase, however, did not reach statistical significance. In conclusion, late stage but not early stage T cell activation antigens were increased in patients with active JCA as compared with patients with JCA in remission and control, whereas some patients in remission had an increased prevalence of T suppressor/inducer cells.     

Two-year evaluation of clinical and laboratory variables of immune function in 117 hemophiliacs seropositive or seronegative for HIV-1

Fifty-nine HIV-1 antibody positive and 58 antibody negative hemophiliacs were evaluated over a 2 year study period to gain insight into the natural history and prognosis of HIV-1 disease in members of this risk group. Mean CD4 (Leu 3+) cell counts calculated at 6 month intervals decreased gradually in seropositive patients (from 403 to 311/microliters) whereas CD8 (Leu 2+) counts remained stable but above the normal range. CD4 cell counts correlated closely with advancing CDC clinical stage; CD8 numbers showed no such association, but were markedly lower in the six patients with overt AIDS. Serum P24 antigenemia was associated with low CD4 cell counts and with advanced clinical stage (58% of antigenemic and 14% of non-antigenemic seropositive patients were in stage IV). In addition to CD4 cell counts, significant reductions in Leu 11+ natural killer cell (NK) subsets and in Leu 3 + 8 - cells occurred in seropositive patients over the study period; Leu 2 + DR + cells increased significantly. When expressed as a percentage of lymphocytes, the reduction in Leu 19 + NK cells was also significant, as were the increases in Leu 4 + DR + cells and Leu 12 + 8 + B cells. In summary, declining CD4 cell numbers and percentages are valuable markers of progressive HIV-1 disease in hemophiliacs, but may not always accurately reflect the degree of disease activity. Progressive changes in additional variables such as serum P24 antigen, and numbers and percentages of NK cell subsets and (as AIDS supervenes) CD8 cell numbers, may allow more precise monitoring of HIV-1 disease. This will, in turn, facilitate the design of optimal individualized strategies for therapeutic intervention.     

Two-dimensional flow cytometric analysis of intrahepatic lymphocyte subsets from patients with chronic hepatitis

Peripheral and intrahepatic lymphocyte subsets were analyzed in 22 patients with chronic hepatitis by two-dimensional flow cytometry. Activated T cells in the liver significantly increased compared with those in the peripheral blood. Helper T cells increased, but the CD4+ cells decreased due to a marked decrease of suppressor inducer T cells. CD8+ cells increased due to a increase of both cytotoxic T and suppressor T cells. Fc-receptor-positive cells, which increased significantly, were not NK cells but Fc-receptor-bearing T cells. In comparison with immunohistochemical methods, flow cytometric analysis enables more objective quantitation and simpler two-color staining of intrahepatic lymphocytes. Our findings using this method suggest that activated T cells and helper T cells have important roles in hepatitis and that hepatocellular injury may be generated not only by cytotoxic T cells but also by Fc-receptor-bearing T cells.     

Learning from lesions: patterns of tissue inflammation in leprosy.

The clinical forms of leprosy constitute a spectrum that correlates closely with the degree of cell-mediated immunity. Patients with tuberculoid leprosy develop strong cell-mediated responses and have only a few, localized lesions, whereas patients with multibacillary lepromatous leprosy are specifically unresponsive to antigens of Myobacterium leprae. T cells of the CD4+ subset predominate in tuberculoid lesions, whereas CD8+ cells predominate in lepromatous lesions. Monoclonal antibodies that distinguish subpopulations of CD4+ and CD8+ cells were used to analyze the distribution of T cells infiltrating lesions across the disease spectrum. In lepromatous lesions, T cells of T-suppressor phenotype (9.3-) were the predominant CD8+ cells and suppressor/inducer cells (2H4+, Leu-8+) represented half of the CD4+ subset. In tuberculoid lesions, helper T cells (CD4+4B4+) outnumbered suppressor/inducer T cells by 14:1, compared with a ratio of 1.2:1 in peripheral blood. Analysis of the precursor frequency of antigen-reactive T cells permitted us to estimate that there was a 100-fold enrichment of T cells able to proliferate in response to M. leprae antigens in tuberculoid lesions (2/100), when compared with blood from the same patients. The methods used here to characterize the T-lymphocyte subsets and frequency of antigen-reactive T cells in leprosy may be useful in analyzing immunological reactions occurring in lesions of other inflammatory and autoimmune diseases.     

Reduction in the suppressor-inducer T cell subset and increase in the helper T cell subset in thyroid tissue from patients with Graves' disease

The expression of surface markers associated with activation and characterization was compared among T cells in thyroid glands and peripheral blood of 10 patients with Graves' hyperthyroidism receiving chronic antithyroid drug therapy, in peripheral blood of 15 patients with untreated hyperthyroid Graves' disease, and in peripheral blood of 21 normal subjects using two-color flow cytometry. In the chronically treated Graves' disease patients, the percentage of activated T cells (HLA-DR+ T cells) among total T cells was significantly higher in thyroid tissue than in peripheral blood, and the increase in percent activated T cells was also significant among both helper/inducer T cell (CD4+ cell) and suppressor/cytotoxic T cell (CD8+ cell) subsets. The percentage of activated T cells in peripheral blood was not significantly different between chronically treated hyperthyroid Graves' patients and normal subjects, whereas the percentage of activated T cells in the peripheral blood from untreated hyperthyroid Graves' disease patients was significantly higher than that in normal subjects or chronically treated hyperthyroid Graves' patients. The percentages of CD4+ cells and CD8+ cells among total T cells were not different between thyroid tissues and peripheral blood in patients with chronically treated hyperthyroid Graves' disease. When CD4+ were further divided into helper T cells (CD4+2H4- cells) and suppressor-inducer T cells (CD4+2H4+ cells) using two-color flow cytometry, the percentage of helper T cells among CD4+ cells was significantly higher in thyroid tissue than in peripheral blood, resulting in an increased ratio of CD4+2H4- cells to CD4+2H4+ cells. The percentage of CD4+2H4+ cells in peripheral blood, however, was not significantly different among untreated and chronically treated Graves' disease patients and normal subjects. From the findings of abnormalities in intrathyroidal T cell subsets, we suggest that the decrease in the function of suppressor T cells within the thyroids of Graves' disease patients may be due to a decrease in CD4+2H4+ cells within thyroid tissue.     

Increase in activated T cells and reduction in suppressor inducer T cells in systemic sclerosis.

Blood lymphocytes from 37 patients with systemic sclerosis were characterised using monoclonal antibodies in a two colour flow cytometric (fluorescence activated cell sorter (FACS)) analysis. The ratio of helper CD4+ to suppressor/cytotoxic CD8+ T cells was raised in patients compared with that in 30 healthy controls owing to decreased CD8+ cells. In the patients CD4+ and CD8+ cells displayed an increased expression of the activation marker HLA-DR. The relative number of CD11b+ CD8+ lymphocytes (suppressor T cells) was normal, but the calculated absolute counts of this cell type were slightly reduced. The proportions and absolute numbers of suppressor inducer T cells, defined as CD45R+ CD4+ cells, were on average only half the levels observed in controls. These findings were not related to the inflammatory activity as measured by acute phase plasma proteins or serum immunoglobulins. Activated T cells were seen at all stages of the sclerotic process and especially during the early stages of the disease and in patients who had suffered occupational exposure to silica dust. A high proportion of activated T cells was also linked with impaired small intestine function but not with the degree of skin or lung involvement. A loss of suppressor inducer T cells was more pronounced later in the disease and in patients with the CREST (calcinosis, Raynaud's phenomenon, oesophageal dysmotility, sclerodactyly, telangiectasia) syndrome. These data provide further evidence for an involvement of T cell mediated immunity in the perpetuation of systemic sclerosis.