concanavalin-a and Liver-Diseases--Alcoholic

Altered immunity is commonly associated with alcoholism. However, few studies have contrasted alcoholism per se with effects of the medical sequelae or comorbidities of alcoholism on the immune system. We previously found few differences in lymphocyte subsets, mitogen response, granulocytic phagocytosis, or natural killer cell activity when we compared healthy urban alcohol-dependent individuals with community control subjects. To begin to explore the role of medical factors, 11 alcohol-dependent persons derived from the same clinical population but showing mild medical abnormalities (AMMAs), primarily abnormal liver function test results, were compared with the previously described 44 alcohol-dependent persons without medical dysfunctions and 34 nonabusing community persons. The AMMAs had lower numbers of CD45RA + inducer-suppressor/naive cells (P <.02) and HLA-DR+-activated T cells (P <.04) compared with findings for nonabusers and higher percentages of circulating CD56 + natural killer cells (P <.03). Mitogen responses to concanavalin A, phytohemagglutinin, and pokeweed mitogen; natural killer cell activity; and granulocyte functions did not differ across groups. The AMMAs reported higher alcohol consumption than that reported by the other groups. The findings seem to indicate that mild medical conditions, or conditions linked to abnormal liver function test results, are associated with some of the immune alterations reported in alcohol-dependent persons. Immune changes, even among chronically alcohol-dependent persons, may occur along a continuum associated with total alcohol exposure and intercurrent physiologic abnormalities. Clinical studies may need to control for such mild abnormalities when investigating alcohol-immune relationships, and clinical interventions may be especially important for alcohol-dependent individuals who show early signs of compromised health.

Topics: Adolescent; Adult; Aged; Alcoholism; CD56 Antigen; Concanavalin A; Female; Granulocytes; Health Status; HLA-DR Antigens; Humans; Immunity; Killer Cells, Natural; Leukocyte Common Antigens; Leukocyte Count; Liver Diseases, Alcoholic; Lymphocyte Count; Male; Middle Aged; Phytohemagglutinins; Pokeweed Mitogens; T-Lymphocytes; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

In previous reports we have demonstrated high plasma levels of sex hormone-binding globulin (SHBG) in asymptomatic alcoholic men. In the present work the physicochemical properties of SHBG from plasma of noncirrhotic alcoholic patients have been further compared with SHBG of control subjects. Steroid binding to SHBG was similar for the two groups: alcoholic men, K(d) of 0.62 +/- 0.07 nM and control individuals, K(d) of 0.70 +/- 0.10 nM. The structure of oligosaccharides attached to SHBG from controls and alcoholic men were determined by using serial chromatography. Our data indicated that 7% of SHBG of control individuals was not retarded by the Con-A column, whereas approximately 30% of SHBG of alcoholic men eluted in the void volume of Con A. Approximately 46% of SHBG of alcoholics applied to Con A, possessed biantennary complex oligosaccharides, as indicated by the fact that it could be eluted with methyl-alpha-D-glucopyranoside and by its retention on wheat germ agglutinin; in contrast, when SHBG from control men was analyzed, approximately 51% was eluted with methyl-alpha-D-glucopyranoside. Approximately 9% of the biantennary complex oligosaccharides on SHBG of control men and none of those on SHBG from alcoholic men were fucosylated on the chitobiose core, as determined by chromatography on Lenn culinaris lectin. Galactosylated oligosaccharides were also present on the SHBG fraction as indicated by its interaction with Ricinus communis-I. Approximately 24% of SHBG of alcoholic men and 39% of those on SHBG from control individuals applied to Con-A were retained and could be eluted with methyl-alpha-D-mannopyranoside. Evidence based on the binding on mannoside-eluted SHBG to Con-A, wheat germ agglutinin, and R. communis-I indicated that at least the SHBG in this fraction, from alcoholics or controls, contained two glycosylation sites and that the sites were differentially glycosylated.

Topics: Adult; Chemical Fractionation; Chromatography, Affinity; Chromatography, Agarose; Concanavalin A; Dihydrotestosterone; Dose-Response Relationship, Drug; Humans; Liver Diseases, Alcoholic; Male; Middle Aged; Oligosaccharides; Protein Binding; Sex Hormone-Binding Globulin; Tritium

Administration of concanavalin A (Con A) leads to acute hepatitis that involves T-cell activation and inflammatory mediator production in mice and rats. We examined the role of CH-100, a Chinese herbal medicine previously trialed in human hepatitis C, in the prevention of Con A-related, T-cell-mediated, acute liver injury in rats.. Female Wistar rats were fed 40% ethanol, 2% sucrose, or isocaloric sucrose for 8 weeks. At the same time, these animals were fed either the Chinese herbal medicine CH-100 (4 tablets/kg body weight/ day) or placebo in chow daily. Blood from the tail vein was collected for endotoxin (lipopolysaccharide) assay at 0, 4, and 8 weeks of ethanol consumption. Twenty-four hours after injection of Con A (20 mg/kg body weight) or phosphate-buffered saline, blood from the tail vein was collected for alanine aminotransferase and tumor necrosis factor (TNF)-alpha assays. Liver-associated CD4+ T cells were isolated from liver perfusates and then cultured with Con A (5 microg/ml) at 37 degrees C for 24 hr. Supernatants were harvested for TNF-alpha assay. The proportion of CD4+ T cells in blood and liver perfusates was measured. Liver samples were collected for histopathological analysis.. Lipopolysaccharide levels were significantly reduced in CH-100-treated ethanol-fed rats compared with placebo-treated rats. After Con A injection, alanine aminotransferase levels were lower at 12 and 24 hr in herb-treated rats compared with placebo-treated rats. Furthermore, serum TNF-alpha levels were lower in ethanol-fed rats on herbal treatment. A significant decrease in TNF-alpha production by liver-associated CD4+ T cells in culture was observed in CH-100-treated ethanol-fed rats. CH-100 treatment was associated with a decreased percentage of CD4+ cells in both blood and liver perfusate in all groups. Herb-treated rats displayed markedly less hepatic necrosis and a reduced CD4+ T-cell infiltrate in portal areas than did placebo-fed rats.. The results demonstrate that CH-100 modified the T-cell response to Con A injection. The effect was more marked in ethanol-fed rats, which suggests a possible role for CH-100 in treating alcoholic liver disease.

Topics: Alanine Transaminase; Animals; CD4-Positive T-Lymphocytes; Central Nervous System Depressants; Concanavalin A; Drugs, Chinese Herbal; Ethanol; Female; Hepatitis, Animal; Humans; Lipopolysaccharides; Liver Diseases, Alcoholic; Mice; Rats; Rats, Wistar; Tumor Necrosis Factor-alpha

It is well-known that microheterogeneity of human serum transferrin observed in alcoholics manifests as sialic acid-deficient transferrin isoforms, otherwise known as carbohydrate-deficient transferrin (CDT). A recent study demonstrated that serum CDT lacked one or both of the entire carbohydrate chains but the investigation required several troublesome procedures. The aim of the present study was to confirm the sugar chain structures of serum transferrin, and of serum CDT in particular, from patients with alcoholic liver disease (ALD) using conventional lectin affinity electrophoresis which might be useful in the clinical setting. The serum CDT obtained from ALD-patients was partially purified using an anion exchanger. Serum transferrin and the partially purified serum CDT were investigated by concanavalin A (Con A)- and Datura stramonium agglutinin (DSA)-affinity electrophoresis followed by antibody-affinity blotting and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with Western blotting. By Con A-affinity electrophoresis, serum CDT was separated into weakly reactive and nonreactive transferrins which showed slower electrophoretic mobilities than those from the healthy controls. Moreover, nearly all of the serum CDT was nonreactive with DSA. On SDS-PAGE, the molecular masses of serum CDT were estimated to be approximately 75 and 72 kDa, which corresponded to those of partially and completely deglycosylated transferrin obtained from the healthy controls (78 kDa), respectively. In conclusion, these results indicated that the sugar chain structures of serum CDT from patients with ALD show not merely a loss of terminal sialic acids, but also the absence of asparagine-N-linked oligosaccharides.

Topics: Antibody Affinity; Blotting, Western; Concanavalin A; Electrophoresis, Agar Gel; Humans; Lectins; Liver Diseases, Alcoholic; N-Acetylneuraminic Acid; Plant Lectins; Protein Conformation; Transferrin

In order to test whether abnormalities in hepatocytes affect the glycoprotein carbohydrate moiety, crossed immunoaffinoelectrophoresis (CIAE) with Concanavalin A (Con A) was used to study serum alpha 1-acid glycoprotein (alpha 1-AGP) and alpha 2-HS glycoprotein (alpha 2-HS) obtained from alcoholic patients with biopsy-proven liver disease. Cirrhotic patients, placed in groups C1, C2 or C3, according to Pugh's classification, were compared to healthy donors (N) and to steatosic non-cirrhotic patients (S). Con A CIAE patterns revealed in group N three subpopulations for alpha 2-HS and four for alpha 1-AGP. Two main results emerged from this study: (1) in the alcoholic groups, the proportions of Con A-unreactive subpopulations of both glycoproteins increased. Moreover, group N could be separated from group S and group S from all the cirrhotic groups. (2) There was a good correlation between the relative amounts in Con A-unreactive subpopulations of alpha 1-AGP and alpha 2-HS. The increases observed in Con A-unreactive subpopulations are probably a general phenomenon related to alterations in glycosylation processing during liver cell damage.

Topics: Adult; Aged; alpha-2-HS-Glycoprotein; Blood Proteins; Concanavalin A; Female; Humans; Immunoelectrophoresis, Two-Dimensional; Liver Diseases, Alcoholic; Male; Middle Aged; Orosomucoid

Concanavalin A-induced lymphocyte proliferation was studied in 25 patients with alcoholic hepatitis or compensated alcoholic cirrhosis. Nine alcoholics without evidence of liver disease were also evaluated. A nonlinear correlation equation, which was natural logarithmic, was applied to individual dose-response proliferation curves and permitted comparisons between patient groups and controls. The proliferative response in all patient groups was significantly lower when compared to healthy controls and was independent of the presence or absence of liver disease. This suggests that some changes in immune function observed in alcoholics may be linked to the direct effects of alcohol on the immune system rather than to the associated liver disease.

Topics: Alcoholism; Concanavalin A; Female; Hepatitis, Alcoholic; Humans; Liver Cirrhosis, Alcoholic; Liver Diseases, Alcoholic; Liver Function Tests; Lymphocyte Activation; Male

To evaluate the influence of chronic alcohol consumption on the cellular immune system in man, we investigated the immune response to seven intradermally applied common antigens and in vitro stimulation of peripheral lymphocytes by Phytohemagglutinin A and concanavalin A in chronic alcoholics with (n = 15) and without (n = 15) liver disease. The results suggest that the diminished cellular immune response in chronic alcoholics is not primarily a direct sequel to alcohol consumption; it is more likely that the impaired immune response is linked to severity of the liver damage itself and to malnutrition.

Topics: Adult; Aged; Alcoholism; Concanavalin A; Female; Humans; Immunity, Cellular; Intradermal Tests; Liver Diseases, Alcoholic; Lymphocyte Activation; Male; Middle Aged; Nutrition Disorders; Phytohemagglutinins

The immune regulatory T cell status of patients with severe alcoholic liver disease (ALD) was investigated. Using monoclonal antibodies to identify lymphocyte subsets in 22 patients, a significant decrease in the percentage of T suppressor/cytotoxic cells (P less than 0.01) and increase in the percentage T helper/inducer population (P less than 0.05) was observed when the results were compared with 20 normal controls. However, when absolute numbers of these lymphocyte subsets were calculated the patient group did not differ significantly from the controls. Further studies revealed T immunoregulatory cell function to be normal. Concanavalin A induced suppressor cells resulted in equivalent inhibition of autologous cell mitogen responsiveness in the patient and control groups. In addition, purified patient T lymphocytes were demonstrated to provide normal help to and manifest normal suppression of IgG, IgA and IgM synthesis by allogeneic B cells. When spontaneous immunoglobulin synthesis by circulating mononuclear cells was investigated, a significant increase in IgA synthesis was found in the ALD patients (P less than 0.05). These results suggest that T cell immunoregulation is normal in patients with ALD and a defect in this system is not responsible for the increased synthesis of immunoglobulin observed in ALD.

Topics: Adult; Aged; B-Lymphocytes; Cells, Cultured; Concanavalin A; Enzyme-Linked Immunosorbent Assay; Humans; Immunoglobulin A; Immunoglobulin G; Immunoglobulin M; Leukocyte Count; Liver Diseases, Alcoholic; Middle Aged; T-Lymphocytes, Cytotoxic; T-Lymphocytes, Helper-Inducer; T-Lymphocytes, Regulatory

To examine the possible contribution of cellular immunoregulatory mechanisms to the pathogenesis and progression of alcoholic liver diseases, suppressor T-cell function was evaluated in patients with severe active or inactive alcoholic liver disease and compared with normal control subjects. Suppressor T-cell activity after in vitro induction by concanavalin A or other T cell-mediated immune reactions was then assessed. T-cell interactions studied included the proliferative responses of both autologous or allogeneic responding peripheral blood mononuclear cells to T-cell mitogens and to allogeneic cells. No significant differences were found in the ability to induce suppressor T cells between controls and patients with inactive alcoholic liver disease (p < 0.05). In contrast, T cells from patients with severe active alcoholic liver disease failed to develop suppressor cell activity (p < 0.001) after concanavalin A stimulation; restoration of suppressor cell function was found after these patients improved clinically. Adherent cells (monocyte-macrophages) from patients with active alcoholic liver diseases exhibited normal support of concanavalin A-induced blastogenesis and suppressor T-cell generation. This immunoregulatory cell abnormality could be important in the pathogenesis and/or progression of active alcoholic liver disease.

Topics: Adult; Concanavalin A; Female; Humans; Immunity, Cellular; Liver Diseases, Alcoholic; Lymphocyte Activation; Male; Middle Aged; T-Lymphocytes, Regulatory

Elevated serum immunoglobulin levels are a frequent finding in patients with severe alcoholic liver disease. We measured in vitro peripheral blood lymphocyte IgA synthesis by radioimmunoassay in 13 patients with severe alcoholic liver disease and hypergammaglobulinaemia and in normal control subjects. We found an increase in spontaneous IgA synthesis compared to controls (P greater than 0.001). On the other hand, there was no difference between groups in percentage stimulation of IgG production by B cells exposed to pokeweed mitogen. We also searched for defects in concanavalin A (Con A) induced suppressor T cell activity. There was no difference between patients with alcoholic liver disease and controls in the capability of such suppressor T cells to inhibit the response of allogeneic cells to a T cell mitogen. Similarly, we examined the capability of Con A-induced suppressor T cells to inhibit pokeweed mitogen-stimulated IgG synthesis by both autologous and allogeneic responder cells and observed no difference between alcoholic patients and controls. Thus these measured suppressor T-T and T-B cell interactions appeared no different from those in control subjects. Our studies suggest, therefore, that B cell IgG synthesis in vitro is enhanced in alcoholic liver disease. Furthermore, the capability to induce suppressor T cells which affects both T and B cell function appears intact; a finding in contrast to our previous observation in chronic active hepatitis. The investigations suggest that enhanced spontaneous IgG synthesis in vitro may result from intense antigenic stimulation in vivo.

Topics: Concanavalin A; Humans; Immunoglobulin G; Liver Diseases, Alcoholic; Lymphocyte Activation; Lymphocytes; Pokeweed Mitogens; T-Lymphocytes, Regulatory

Mallory bodies (MBs) were obtained in purified form from human liver obtained at autopsy using a new procedure consisting of sedimentation through a Ficoll viscosity barrier. Preparations from six livers ranged in purity from 95 to 99 per cent. MB preparations were autofluorescent. MBs were strongly agglutinated by Concanavalin A. The presence of carbohydrate was also indicated by the fact that MBs bound fluorescently labeled Concanavalin A; no binding was observed in the presence of appropriate inhibitor monosaccharides. Direct analysis indicated that MBs contained variable amounts of neutral hexose (0.65 to 2.4 mumoles of glucose-equivalents per milligram of protein) but no sialic acid. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that purified MBs contain five major polypeptides possessing apparent molecular weights of 56,000, 48,500 to 45,000 (triplet), and 32,500. Periodic acid-Schiff-positive components were not detected. Scanning electron microscopy of isolated MBs revealed the presence of a rough, fibrous surface, whereas conventional transmission electron microscopy indicated the filamentous nature of MBs.

Topics: Concanavalin A; Electrophoresis, Polyacrylamide Gel; Fluorescent Antibody Technique; Hexoses; Humans; Hyalin; Inclusion Bodies; Liver; Liver Diseases, Alcoholic; Microscopy, Electron, Scanning; Peptides