concanavalin-a and Liver-Cirrhosis--Biliary

The familial occurrences of biochemical and immunological abnormalities and histocompatibility antigens were studied in 18 healthy first-degree relatives of patients with primary biliary cirrhosis (PBC) in two families. In each of these two families, there were two members who suffered from PBC. All relatives had normal serum aspartate aminotransferase, alkaline phosphatase, bilirubin, total cholesterol, and immunoglobulins except the two, who had a mild elevation of alkaline phosphatase without cholestasis. Autoantibodies were present in some relatives; five (28%) for antithyroglobulin antibody and antithyroid microsomal antibody, one (6%) for antimitochondrial and antinuclear antibody, and one (6%) for rheumatoid factor. Abnormalities of T or B lymphocytes in peripheral blood were detected in two (11%) relatives. Impairment of concanavalin A-induced lymphocyte transformation determined by ethidium bromide fluoroassay was found in seven (39%) relatives, although an abnormal response for phytohemagglutinin was detected in none of the relatives. The HLA haplotypes were not necessarily associated with positive autoantibodies or impaired concanavalin A-induced lymphocyte transformation in these families. These findings suggest that impairment of concanavalin A-inducible lymphocytes (mainly suppressor T cells) is one of the contributing factors in the development of PBC.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Autoantibodies; B-Lymphocytes; Concanavalin A; Family; Female; HLA Antigens; Humans; Leukocyte Count; Liver Cirrhosis, Biliary; Lymphocyte Activation; Male; Middle Aged; Pedigree; Rosette Formation; T-Lymphocytes

Lymphocyte and monocyte function was investigated in eight patients with primary biliary cirrhosis and in age-matched and sex-matched controls. In three of the eight cirrhotic patients, two were in the late stage of the disease (stage III) and concanavalin A-induced suppressor cell function was markedly decreased; it returned to normal levels after 1 mo of treatment consisting of 5 mg per week of orally administered colchicine. The relative percentage of T cell subsets (CD3, CD4 and CD8) and the mean percentage of the helper and the suppressor phenotype were similar in cirrhotic patients and in controls. Basal production of interleukin-1 activity by a monocyte-enriched fraction of peripheral blood mononuclear cells of cirrhotic patients was three-fold above control values: after stimulation with endotoxin, interleukin-1 values increased tenfold above basal levels in monocyte-enriched fraction of peripheral blood mononuclear cells of both normal and cirrhotic patients. Colchicine normalized the basal production of interleukin-1 and inhibited its stimulation by endotoxin by 50%. Culture supernatants of endotoxin-stimulated monocyte-enriched fraction of peripheral blood mononuclear cells increased the proliferation of cultured human skin fibroblasts by 50%. In contrast, supernatants from stimulated monocyte-enriched fraction of peripheral blood mononuclear cells obtained from colchicine-treated patients significantly inhibited fibroblast proliferation. These findings suggest that stimulated chronic inflammatory cells may play an important role in liver fibrosis. Some of the actions of colchicine could be related to its effects on lymphocyte and monocyte function.

Topics: Adult; Aged; Cell Division; Colchicine; Concanavalin A; Fibroblasts; Humans; Interleukin-1; Liver Cirrhosis, Biliary; Lymphocytes; Middle Aged; Monocytes; T-Lymphocytes, Regulatory

Suppressor cell function was studied in 31 patients with primary biliary cirrhosis. Blood mononuclear cells were activated in vitro with suboptimal or optimal concentrations of concanavalin A and suppression of mitogen-induced proliferation or synthesis of anti-mitochondrial antibodies was measured. At suboptimal concentrations of concanavalin A, mean (+/- SE) suppression of proliferation by patients' cells was significantly less than 14 controls (18.7 +/- 5% vs 34.3 +/- 5%; P less than 0.005). The degree of suppressor activity correlated with the clinical disease, i.e., suppression was greater in patients with early than advanced disease. Spontaneous anti-mitochondrial antibody synthesis was not detected in cultures of mononuclear cells from normal subjects nor from patients with negative serum antibody, nor could it be induced by pokeweed mitogen in either group. In contrast, spontaneous anti-mitochondrial antibody synthesis was detected in cultures of 18/23 patients with positive serum antibody and was significantly augmented by pokeweed in 12/18. Anti-mitochondrial antibody synthesis was augmented by 48% (+/- 19) in response to pokeweed and concanavalin A-induced suppression higher (39.4 +/- 7%) in patients with low titer antibody compared to 1.6% (+/- 8) pokeweed augmentation and 5.5 +/- 3% concanavalin A-induced suppression in patients with high titer antibody. These results suggest that some patients with primary biliary cirrhosis have an immunoregulatory defect which expresses itself in part as an inability to regulate autoantibody synthesis.

Topics: Antibody Formation; Autoantibodies; Colchicine; Concanavalin A; Female; Humans; In Vitro Techniques; Liver Cirrhosis, Biliary; Lymphocyte Activation; Lymphocytes; Male; Mitochondria; T-Lymphocytes, Regulatory

Peripheral blood lymphocytes from patients with primary biliary cirrhosis previously have been reported to demonstrate reduced pokeweed mitogen-stimulated immunoglobulin synthesis and diminished function of suppressor T cells. To determine whether thymic hormone preparations reverse these immunologic defects in vitro, the effects of thymosin fraction 5 and thymosin alpha 1 on immunoglobulin synthesis and concanavalin A-induced suppression of immunoglobulin synthesis were investigated in normal subjects and patients with primary biliary cirrhosis. In normal subjects, no effects of thymosin were observed on unstimulated and pokeweed mitogen-stimulated immunoglobulin synthesis, nor on Con A-induced suppressor cell activity. Lymphocytes from patients with PBC synthesized less IgG and IgM than normals when stimulated by pokeweed mitogen, and this difference was enhanced by both thymosin fraction 5 and thymosin alpha 1. Con A suppression of immunoglobulin synthesis was abnormal in only one PBC subject so that thymosin effects on impaired suppressor T cell activity could not be tested.

Topics: Adult; Aged; Cells, Cultured; Concanavalin A; Female; Humans; Immunoglobulin G; Immunoglobulin M; Liver Cirrhosis, Biliary; Lymphocytes; Male; Middle Aged; Pokeweed Mitogens; T-Lymphocytes, Regulatory; Thymalfasin; Thymosin

The in vitro effect of Cyclosporin A on the regulation of immunoglobulin production was investigated in 16 patients with primary biliary cirrhosis. A significant improvement in concanavalin A induced suppression of IgG and IgM producing cells was observed after prior incubation of mononuclear cells with 300 ng/ml Cyclosporin A for 30 minutes. No effect was seen on spontaneous or pokeweed mitogen induced immunoglobulin production, nor on con A induced suppression if Cyclosporin A was added after 24 hours. Incubation of mononuclear cells with a variable dose of Cyclosporin A showed an effect only at 250-500 ng/ml. Higher and lower doses had no effect. This dose dependent effect of Cyclosporin A is likely to be related to a differential inhibitory effect on T helper and T suppressor cells and may underlie the clinical benefit being observed in current clinical trials.

Topics: Aged; Cells, Cultured; Concanavalin A; Cyclosporins; Female; Humans; Immunoglobulin G; Immunoglobulin M; Liver Cirrhosis, Biliary; Male; Middle Aged; T-Lymphocytes, Regulatory

Immunoglobulin production and its regulation have been investigated in 36 untreated patients with primary biliary cirrhosis and 14 healthy controls. Using a haemolytic plaque assay, the number of cells spontaneously producing IgG or IgM was normal, while in contrast, proliferation of both IgG and IgM producing cells in the presence of pokeweed mitogen was impaired (P less than 0.01). There was in addition a defect of Con A induced suppression of proliferation of IgG and IgM producing cells (P less than 0.01), which was more marked in those with early disease than in those with advanced disease. In vitro incubation of mononuclear cells with prednisolone, 5 X 10(-8) M, corrected the defect of Con A induced suppression, but had no effect on spontaneous or pokeweed mitogen stimulated immunoglobulin production. Prednisolone was only effective if added prior to exposure to Con A. There is anecdotal evidence that corticosteroids may be beneficial in primary biliary cirrhosis and the results of the present study study suggest that further investigation of their role is warranted.

Topics: Adult; Aged; Cells, Cultured; Concanavalin A; Female; Humans; Immune Tolerance; Immunoglobulin G; Immunoglobulin M; Immunoglobulins; Liver Cirrhosis, Biliary; Lymphocyte Activation; Male; Middle Aged; Pokeweed Mitogens; Prednisolone; T-Lymphocytes, Regulatory

To evaluate the mechanisms underlying the increase in serum IgM in primary biliary cirrhosis (PBC) studies were designed to examine IgM production in vitro and to assess the relative contribution of intrinsic B cell activity and immunoregulatory T cell balance to IgM synthesis. The number of peripheral blood lymphocytes (PBL) producing IgM (spontaneous and pokeweed mitogen (PWM) stimulated) at the end of a seven day culture period was similar in PBC patients and control subjects while the amount of IgM synthesized (spontaneous and PWM stimulated) during this period was significantly greater in the patient group, implying that the amount of IgM produced per B cell was increased in PBC. Co-culture of autologous and allogeneic T and B lymphocytes and irradiation of T lymphocytes from patients and normal subjects clearly implicated abnormal suppressor T cell function, rather than autonomous B cell hyperactivity, as the cause of the increased IgM synthesis. Direct studies of T cell function indicated that although concanavalin A (Con A) activated suppressor cells inhibited proliferation of IgM producing B cells in the majority of PBC patients, they were unable to inhibit IgM synthesis. The demonstration of a disparity between IgM synthesis and the proliferation of IgM-producing B cells, together with the observation that the abnormality of T cell function is largely confined to the control of IgM secretion, is consistent with the presence of at least two different suppressor subpopulations regulating IgM production. In PBC the main suppressor cell abnormality seems to affect regulation of IgM secretion rather than B cell proliferation.

Topics: B-Lymphocytes; Cells, Cultured; Concanavalin A; Humans; Immunoglobulin G; Immunoglobulin M; Liver Cirrhosis, Biliary; Mitosis; T-Lymphocytes; T-Lymphocytes, Regulatory

Patients with primary biliary cirrhosis (PBC) have been previously reported to have immunoregulatory abnormalities. We tested the effect of in vitro colchicine on PBC patients' suppressor cell function in order to determine whether colchicine can correct their suppressor cell deficiency. PBC patients' mononuclear cells were cultured for 44 h with concanavalin A (Con A) as well as with or without colchicine at a pharmacological concentration (10(-8)M) or at a suprapharmacological concentration (10(-5)M) and then tested for their ability to suppress proliferation of phytohaemagglutinin stimulated healthy volunteers' mononuclear cells. Eleven PBC patients had significantly (P less than 0.001) decreased suppressor cell function (12 +/- 15%, mean +/- s.d.) as compared to 37 healthy volunteers (43 +/- 12%). The suprapharmacological concentration of colchicine did not significantly affect the PBC patients' suppressor cell function (16 +/- 15%). In contrast, in the nine PBC patients tested with the pharmacological concentration of colchicine, their suppressor cell function was increased to 40 +/- 20% which was significantly different than without colchicine (P less than 0.01) or with the suprapharmacological concentration of colchicine (P = 0.02) but not significantly different than healthy volunteers. Thus, in vitro colchicine at a pharmacological concentration corrects PBC patients' deficiency of Con A-induced suppressor cell function raising the possibility that oral colchicine might be clinically useful as an immunomodulating drug in PBC.

Topics: Aged; Colchicine; Concanavalin A; Female; Humans; Liver Cirrhosis, Biliary; Middle Aged; T-Lymphocytes, Regulatory

Both concanavalin A (Con A)-induced suppressor cell activity and autologous mixed lymphocyte reaction (AMLR) were significantly decreased in patients with primary biliary cirrhosis (PBC) in comparison with those of healthy subjects (p less than 0.001) and p less than 0.001, respectively). Factors inhibiting lymphocyte transformation by Con A and PHA were demonstrated in sera from a majority of the patients with PBC. In approximately half of these patients, some serum factor(s) also induced a decrease of Con A-induced suppressor cell function of lymphocytes from healthy subjects. These results suggest that patients with PBC have impaired immunoregulatory lymphocyte function which may be related to some factor(s) present in serum.

Topics: Adult; Aged; Concanavalin A; Female; Humans; Liver Cirrhosis, Biliary; Lymphocyte Activation; Lymphocyte Culture Test, Mixed; Male; Middle Aged; Phytohemagglutinins; T-Lymphocytes; T-Lymphocytes, Regulatory

Patients with primary biliary cirrhosis have an abnormality of lymphocyte suppressor cell function that could play a role in the pathogenesis of their disease. To investigate this possibility, suppressor cell function in patients with primary biliary cirrhosis, in their healthy first-degree relatives, in unrelated household contacts, in patients with other types of cirrhosis, and in normal control subjects was studied. The method used is based on the finding that the in vitro addition of concanavalin A to pokeweed mitogen-stimulated lymphocytes activates suppressor cells, which in turn inhibit immunoglobulin synthesis. Thirteen of 16 patients with primary biliary cirrhosis and six of 23 healthy relatives had significant impairment of IgG suppression. All six of these relatives were female. No abnormal suppression was found in unrelated household contacts of patients with primary biliary cirrhosis, in patients with other types of cirrhosis, or in healthy control subjects. There was no correlation between results of the IgG suppressor cell assay and the disease activity in patients with primary biliary cirrhosis. These data suggest that the abnormal suppressor cell function is not a direct cause of primary biliary cirrhosis; however, it may be a genetic marker for susceptibility to this disease.

Topics: Adult; Concanavalin A; Female; Genetic Markers; Humans; Immunoglobulin G; In Vitro Techniques; Liver Cirrhosis, Biliary; Male; Middle Aged; Pokeweed Mitogens; T-Lymphocytes, Regulatory

To evaluate the interactions between circulating immune complexes (CIC) and lymphoid cells in primary biliary cirrhosis (PBC), we determined (1) whether antibodies to lymphocytes in PBC serum, independent of CIC, could account for binding in the Raji cell assay for CIC and (2) whether CIC or other humoral factors in PBC serum could interact with lymphoid cells to alter their function. We found that three quarters of CIC positive PBC sera bound specifically to Raji cells via complement receptors, while only one quarter had antibodies to lymphoid cells or Raji cells devoid of complement receptors. We also demonstrated factors which inhibited cell-mediated cytotoxicity and suppressor cell activity in PBC sera; however, we found no correlation between the level and presence of CIC or of lymphocyte antibodies and the level or presence of these serum inhibitory factors. Thus, although the detection of CIC in PBC is not artifactual, the contribution of CIC and other serum factors to the other immunological aberrations in PBC remains to be elucidated.

Topics: Antibodies, Neoplasm; Antigen-Antibody Complex; Antilymphocyte Serum; Burkitt Lymphoma; Concanavalin A; Cytotoxicity, Immunologic; Female; Humans; Liver Cirrhosis, Biliary; Lymphocytes; T-Lymphocytes, Regulatory

Recent evidence has suggested that peripheral blood suppressor cell populations can modulate immune responsiveness. Absence of suppressor cell activity has been noted in diseases of presumed autoimmune basis. We determined inducible suppressor cell activity in patients with primary biliary cirrhosis (PBC) as compared to age and sex-matched controls. There was a significant loss of suppressor cell activity for mitogen response in patients with PBC (-4.5 +/- 8.0%) versus controls (43.7 +/- 7.8%). No correlation of this loss to clinical parameters was observed. This study suggests PBC patients lack inducible suppressor cell activity provide partial explanation for the perpetuation of this disease.

Topics: Adult; Bilirubin; Concanavalin A; gamma-Globulins; Humans; Immunoglobulin M; Liver Cirrhosis, Biliary; Lymphocyte Activation; Middle Aged; Mitomycins; T-Lymphocytes, Regulatory

An additional cholestatic lipoprotein with a slower mobility than the usual alpha-lipoprotein on polyacrylamide-gel disc-electrophoresis was found in the serum of patients with cholestasis. This abnormal lipoprotein was referred to as Slwo-migrating HDL (HDL-S), because it was mostly recovered in the high density lipoprotein (HDL) fraction after preparative ultracentrifugation. HDL-S was precipitated by dextran sulfate and Mg++ but did not react with either concanavalin A or anti-beta-lipoprotein serum. The main apoprotein of HDL-S was Apo A-I, and a trace of Apo E was also present. HDL-S was relatively enriched in free cholesterol and triglycerides and had a density in the range of 1.063 to 1.083. The appearance of HDL-S in serum or plasma was closely associated with chronic mild intrahepatic cholestasis, particularly as in primary biliary cirrhosis and related conditions.

Topics: Cholestasis, Intrahepatic; Concanavalin A; Dextrans; Electrophoresis, Polyacrylamide Gel; Humans; Lipoproteins, HDL; Liver Cirrhosis, Biliary; Magnesium; Ultracentrifugation