In vitro responses to a 65-kilodalton mycobacterial protein by synovial T cells from inflammatory arthritis patients.

Bacterial Ag, especially those of mycobacteria, have been implicated in the pathogenesis of experimental inflammatory arthritis in rodents, while in man, reactive arthritis has a clear temporal relationship to infection with particular bacteria. To investigate the role of immune responses to bacterial Ag in inflammatory arthritis, we have examined the proliferative responses of paired synovial fluid and PBMC when stimulated with 1) suspensions of irradiated or heat-killed bacteria associated with reactive arthritis (ReA), 2) purified protein derivative, 3) a recombinant 65-kDa heat shock protein of Mycobacterium leprae. The 65-kDa Ag was stimulatory to synovial fluid mononuclear cells, but not PBMC, from patients with different arthropathies, including most of those with ReA, but also some with rheumatoid arthritis.

Synovial T lymphocyte recognition of organisms that trigger reactive arthritis.

Reactive arthritis (ReA) is believed to be "triggered' by infection with certain bacteria. When the proliferative responses of mononuclear cells (MC) obtained from the synovial fluid (SF) of ReA patients were examined, it was found that they responded maximally to the specific organism responsible for the preceding infection. The response was shown to be due to Class II MHC-restricted T cells by inhibition experiments using cyclosporin A and monoclonal antibodies.

Interleukin-2 production and response by helper T-cell subsets in man.

This study investigated the production of, and response to, the lymphokine interleukin-2 (IL-2) by functional subsets of human CD4+ T lymphocytes. Fresh peripheral blood mononuclear cells (PBMC) were sorted into CD4+2H4+/4B4- suppressor-inducer cells, and CD4+2H4-/4B4+ helper cells. The suppressor-inducer subset proliferated well in response to the T-cell mitogens concanavalin A (Con) A and phytohaemagglutinin (PHA), and produced IL-2.

Deficient interleukin 2 production in rheumatoid arthritis: association with active disease and systemic complications.

Interleukin 2 (IL-2) production and proliferative responses of peripheral blood mononuclear cells (PBMC) stimulated with three concentrations of PHA were measured in 75 patients with rheumatoid arthritis (RA) and 25 normal controls. All patients were on a standard therapeutic regime, and were assessed for disease activity by clinical and laboratory criteria. Rheumatoid cells showed significantly lower IL-2 production and proliferation than normal PBMC at all PHA doses.

Acute confusional state and hyponatraemia due to inappropriate antidiuretic hormone secretion in polyarteritis nodosa.

An acute confusional state, which developed in a patient with polyarteritis nodosa (PAN), proved to be secondary to inappropriate secretion of antidiuretic hormone and consequent hyponatraemia. This is a very unusual complication of PAN and may well reflect a direct stimulation of the supraoptic nuclei owing to cerebral vasculitis.

Dissection of the mechanisms of immune injury in rheumatoid arthritis, using total lymphoid irradiation.

Eleven patients with intractable rheumatoid arthritis were treated with total lymphoid irradiation. After radiotherapy, there was a marked decrease in the number and function of peripheral blood helper/inducer (Leu-3+) T lymphocytes, in the spontaneous secretion of interleukin-1 by synovial biopsy specimens, and in the activity of the joint disease. In contrast, levels of IgM, IgA, and IgG rheumatoid factors and C3 concentrations in blood and synovial fluid samples did not change significantly after therapy with total lymphoid irradiation.

Enhancement of human T-lymphocyte growth by human transferrin in the presence of fetal bovine serum.

All dividing cells require transferrin as a growth factor. During in vitro culture of human lymphocytes, transferrin is usually supplied in the form of serum, either synergic or xenogenic (usually fetal bovine serum (FBS)). In the present work the growth of certain human T-cell lines was examined; these lines were derived from the synovium of rheumatoid arthritis patients and maintained in 10% FBS and 1% synovial fluid.

The abnormal cytotoxic T cell response to Epstein-Barr virus in rheumatoid arthritis is correlated with disease activity and occurs in other arthropathies.

The cytotoxic T cell response to Epstein-Barr virus as measured by the regression assay was found to be impaired in a group of patients with active rheumatoid arthritis (RA). When these patients responded clinically to treatment with sulphasalazine there was a concomitant increase in the strength of this virus specific T cell response. The suggestion of a correlation between disease activity and impairment in this immune response was borne out in studies of other groups of patients with RA.

Disturbance of the Epstein-Barr virus-host balance in rheumatoid arthritis patients: a quantitative study.

Rheumatoid arthritis (RA), seronegative spondyloarthropathy (SA) and osteoarthritis (OA) patients receiving no steroid or disease-modifying therapy have been monitored, along with healthy controls, for their prevailing level of Epstein-Barr virus (EBV) infection using four independent indices of the EBV-host balance, levels of virus shedding in throat washings as measured by a cord-blood transformation assay of improved sensitivity, frequency of virus-infected B cells in the circulating blood as measured by the rate of 'spontaneous' transformation in limiting dilution cultures, antibody titres to viral antigens, and virus-specific cytotoxic T cell responsiveness as measured in the in vitro regression assay. All four parameters indicated significant disturbance of the virus-host balance accompanying RA, the range of values exhibited by RA patients as a group in each case extending beyond the normal control range in the direction of more active infection. However, observations with SA and OA patients suggested that such a disturbance may not be RA-specific.

The role of HLA antigens in the control of the cytotoxic T-cell response to Epstein-Barr virus: a family study.

Epstein-Barr (EB) virus-specific, HLA-restricted cytotoxic T-cell populations have been generated in vitro from each member of a family by cocultivating peripheral blood mononuclear cells with autologous EB virus-transformed B cells, the resulting effector cells being expanded as interleukin 2-dependent T-cell lines. The cytotoxicity of each of these effector populations was tested on a large panel of EB virus-transformed target cells of known HLA type, so that the particular HLA antigens which acted as restricting elements for each cytotoxic population could be identified. There was a consistent pattern within the family of preference for certain HLA class I antigens as restricting elements of the virus-specific T-cell response.

In vitro analysis of the Epstein-Barr virus: host balance in long-term renal allograft recipients.

Four indices of the EB virus carrier state, for which quantitative in vitro assays now exist, have been monitored in 55 renal allograft recipients under long-term immunosuppression, each patient being tested on a single occasion. By comparison with parallel data from healthy control donors, the results indicate the extent to which virus replication in the throat and virus-infected B cells in the blood are increased as a result of immunosuppression; the concordance between these two independent indices of the level of EB virus infection in vivo, first noted with healthy donors, was again observed within this large group of patients. Immunosuppression also leads to an impairment of virus-specific memory T-cell responsiveness and to an increase in anti-viral antibody titres, but the results show that the level of virus infection prevailing in any one individual cannot be inferred directly from these immunological indices of the virus:host balance.

Mutant HLA-A2 antigens as restricting elements for virus-specific cytotoxic T cells.

Mutants of the EB virus-transformed cell line T5-1 (HLA-A1, 2; B8, 27), bearing well-characterized alterations in HLA-A2 antigen expression and unable to bind the HLA-A2-specific monoclonal antibody BB7.2, have been tested for their susceptibility to EB virus-specific cytolysis using effector T-cell preparations functionally restricted through relevant HLA antigens. Initial experiments first confirmed that the parent line T5-1 was susceptible to cytolysis by both "common" A2-restricted and B27-restricted effector cells.

Cross-reactivity of self-HLA-restricted Epstein-Barr virus-specific cytotoxic T lymphocytes for allo-HLA determinants.

Epstein-Barr (EB) virus-specific cytotoxic T cells, prepared from virus-immune donors by reactivation in vitro and maintained thereafter as IL-2-dependent T cell lines, have been tested against large panels of EB virus-transformed lymphoblastoid cell lines of known HLA type. Whilst the pattern of lysis of the majority of targets was always consistent with HLA-A and HLA-B antigen restriction of effector function, in several cases it was noticed that certain HLA-mismatched targets were also reproducibly lysed. When this "anomalous" lysis was investigated in detail, it was found to be directed against allodeterminants on class I HLA antigens; thus, mitogen-stimulated as well as EB virus-transformed lymphoblasts from the relevant target cell donors were sensitive to the killing, and in each case the lysis could be specifically blocked by monoclonal antibodies to class I HLA antigens.

Epstein-Barr virus-specific cytotoxic T lymphocytes as probes of HLA polymorphism. Heterogeneity of T cell-restricting determinants associated with the serologically defined HLA-A2 antigen.

Epstein-Barr (EB) virus-specific effector T cell lines were established from nine virus-immune donors positive for the serologically defined HLA-A2 antigen; of these, four lines contained a demonstrable A2-restricted cytotoxic component. When these four effector populations were each tested on the same panel of EB virus-transformed lines from 20 HLA-A2-positive individuals, 16 of the target cell lines were consistently killed at levels above 25% of the relevant autologous cell lysis. Cytotoxicity appeared to be mediated through a restricting determinant associated with the 'common A2' antigen that these lines shared; indeed the lysis could be specifically blocked by high concentrations of an HLA-A2-specific monoclonal antibody.

Role of aspiration cytology in the diagnosis and management of mammary lesions in office practice.

Sixteen hundred and eighty breast aspiration specimens obtained from 1410 patients seen in office practice were reviewed. The cytologic diagnosis was unsatisfactory in 230 cases, benign in 1019 cases, atypical in 198 cases, suspicious for malignancy in 102 cases, and malignant in 131 cases. All cases diagnosed as cytologically malignant had a subsequent tissue diagnosis of malignant neoplasm.

Cytotoxic T cell recognition of Epstein-Barr virus-infected B cells. III. Establishment of HLA-restricted cytotoxic T cell lines using interleukin 2.

Epstein-Barr virus (EBV)-specific cytotoxic T cell precursors, present in the circulation of previously infected (seropositive) individuals, have been reactivated in vitro by challenging with autologous EBV-transformed cells, and the reactivated populations subsequently expanded as interleukin 2 (IL2)-dependent cell lines. These lines were dominated by T cells possessing the cytotoxic/suppressor cell surface phenotype and, when tested for effector function in chromium-release assays, demonstrated potent EBV-specific, HLA-A and -B antigen-restricted cytotoxicity even when derived from seropositive donors whose initial cytotoxic response to in vitro reactivation was relatively weak. With all the lines tested from 10 seropositive donors, strong killing of autologous EBV-transformed cells was observed in the absence of any significant lysis of autologous mitogen-stimulated lymphoblasts or of a panel of EBV genome-negative cell lines sensitive to natural killing.