Cell-mediated immunity in acute and chronic hepatitis.

Wands, Jack R.; Perrotto, Joseph L.; Alpert, Elliot; Isselbacher, Kurt J.

doi:10.1172/jci108021

Cited by 99 publications

(26 citation statements)

References 27 publications

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“…We were not able to demonstrate significant inhibitory activity by sera from patients with CAH or AH on cytotoxicity by autologous lymphocytes in contrast to the findings of other investigators (6,10,19,21). As mentioned previously, antibodies to hepatic antigens present in autologous sera (39) may have blocked SCMC and promoted ADCC.…”

Section: Discussioncontrasting

confidence: 99%

“…Unlike several studies of lymphocyte cytotoxicity in CAC (8,17), we were able to demonstrate that lymphocytes from patients with CAC still manifest in vitro cytotoxic capability. The decreased in vitro cytotoxicity shown by patients with CAH on steroid therapy supports the findings of others (6,7,16,19,20), suggesting further that corticosteroids inhibit the effector phase of lymphocyte-mediated cytotoxicity (35). Several mechanisms have been suggested to account for in vitro cell-mediated cytotoxicity in CAH and AH.…”

Section: Discussionsupporting

confidence: 76%

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Lymphocyte cytotoxicity in human liver disease using rat hepatocyte monolayer cultures

Mutchníck

Missirian

Johnson

1980

Clinical Immunology and Immunopathology

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Isolated rat hepatocytes were used to determine the cytotoxicity of peripheral blood mononuclear cells (PBM) from patients with alcoholic liver disease (ALD) and chronic active hepatitis (CAH). The specificity of the cytotoxic effect on liver cells was monitored using rat kidney cells. PBM from patients with CAH and ALD showed increased spontaneous cell-mediated cytolysis (SCMC) for hepatocytes. The SCMC against kidney cells was comparable for the patient groups and controls. Incubation of liver cells with antibody to human liver-specific protein significantly increased the cytotoxic activity of control PBM but did not block cytotoxicity by PBM of patients with CAH. Incubation of PBM from untreated CAH patients with thymosin fraction 5, a polypeptide extract of the thymus gland, significantly lowered cytotoxicity. Our findings suggest that rat liver cells provide a model for studies of cell-mediated immunity in human liver disease and that thymosin fraction 5 decreases the cytotoxic activity of sensitized lymphocytes in CAH.

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Section: Discussioncontrasting

confidence: 99%

Section: Discussionsupporting

confidence: 76%

Lymphocyte cytotoxicity in human liver disease using rat hepatocyte monolayer cultures

Mutchníck

Missirian

Johnson

1980

Clinical Immunology and Immunopathology

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“…The relatively greater sensitivity to the effects of irradiation of the net T cell helper signal (the balance between helper and suppressor signals) in the antigen-induced system, when compared with the PWM-induced system, is similar to the pattern reported in other in vitro antigen-induced antibody systems (15-17, 19, 36 The human immune response, including cellular immune reactivity to HBsAg, has been extensively evaluated by a variety of techniques, including migration inhibition, cytotoxicity assays, and blast transformation after infection with hepatitis B virus (reviewed, 9 and 10). In the convalescent phase after resolution of hepatitis B viral infection, blastogeneic responses to HBsAg have been reported by some (46-48) but not all authors (49). In the papers that report positive results, the stimulation index has been relatively low (usually .5).…”

Section: Resultsmentioning

confidence: 99%

In vitro antigen-induced antibody responses to hepatitis B surface antigen in man. Kinetic and cellular requirements.

Cupps¹,

Gerin²,

Purcell³

et al. 1984

J. Clin. Invest.

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“…The isolation of peripheral blood lymphocytes, the preparation and [51Cr]chromium labeling of Chang cells, lymphocyte blast transformation, and cytotoxicity studies were performed as previously described (19). RESULTS Characterization of m-Con A.…”

Section: Methodsmentioning

confidence: 99%

Mechanism of human lymphocyte stimulation by concanavalin A: role of valence and surface binding sites.

Wands¹,

Podolsky²,

Isselbacher³

1976

Proc. Natl. Acad. Sci. U.S.A.

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A monovalent form of concanavalin A (m-Con A) has been prepared to determine the importance of valence for human lymphocyte surface binding and subsequent lymphocyte stimulation as measured by blast transformation and cytotoxicity. Concanavalin A (Con A) was fragmented by a proteolytic process and the m-Con A derivative was isolated by elution with an ascending D-glucose gradient on a Sephadex CG200 column. The molecular weight of m-Con A was 18,000 by sodium dodecyl sulfate-polyacrylamide electrophoresis. Equilibrium dialysis with a-methyl Dglucoside and subsequent METHODSPreparation of m-Con A. Con A was fragmented by a modification of the method of Evans and Jones (14). For this procedure, Con A (150 mg) was dissolved in 5.0 ml 0.1 M HCI to which 2 mg of porcine pepsin (California Biomedical Research Co.) was added. The mixture was incubated for 120 min at 400, after which the reaction was stopped by adjusting the pH to neutrality with 0.1 M NaOH. Subsequently, 20.0 ml of phosphate-buffered saline containing 0.9 mM CaC12 and 1.05 mM MgCl2 was added, and the precipitate was removed by centrifugation at 20,000 X g for 20 min. Trypsin treated with L-1-tosylamide-2-phenylethyl-chloromethyl ketone (TPCK) (120 mg) and protease type 6 (40 mg; Sigma Chemical Co.) were then added and the mixture was incubated at 400 for 120 min. Finally, the reaction was stopped by cooling on ice and again the precipitate was removed by centrifugation.The cooled reaction mixture was introduced at a flow rate of 6.0 ml/hr to a column (35.0 cm X 1.5 cm) of Sephadex G-200 equilibrated with phosphate-buffered saline. Subsequently, the column was washed with buffer by means of a peristaltic pump at a flow rate of 5.0 ml/hr and 5.0 ml fractions were collected. Protein concentration in the eluant was determined by continuous monitoring at a wavelength of 280 nm. After 24 hr D.glucose was added to the elution buffer in an ascending linear gradient from 0 to 35 mM using a Pharmacia Ultragrad 2000 gradient maker. A flow rate of 6.0 ml/hr was maintained and 3.0 ml fractions were collected. Each fraction was subsequently dialyzed extensively against phosphate-buffered saline and the total protein concentration of the pooled fraction was measured by the method of Lowry et al. (15).After dialysis, each fraction eluted during the application of the linear glucose gradient was assayed for agglutinating ability. Fresh washed rabbit erythrocytes were prepared and agglutination studies were performed as previously described (16). Rabbit erythrocytes were regularly agglutinated at concentrations of 1 ug/ml of untreated Con A. Those fractions obtained after elution with D-glucose (and subsequent dialysis against phosphate-buffered saline) that failed to cause agglutination after 40 min at protein concentrations greater than 10 ,gg/ml were pooled and subjected to further characterization. This pooled fraction was also evaluated for its ability to agglutinate human lymphocytes in a similar manner to the assay for rabbit erythrocyte agglutination. ...

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Cell-mediated immunity in acute and chronic hepatitis.

Cited by 99 publications

References 27 publications

Lymphocyte cytotoxicity in human liver disease using rat hepatocyte monolayer cultures

Lymphocyte cytotoxicity in human liver disease using rat hepatocyte monolayer cultures

In vitro antigen-induced antibody responses to hepatitis B surface antigen in man. Kinetic and cellular requirements.

Mechanism of human lymphocyte stimulation by concanavalin A: role of valence and surface binding sites.

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