Dissociation of hepatitis A virus antigen-anti-HAV antibody complexes by 2-mercaptoethanol and dithiothreitol.
Intravenous inoculation of two marmosets and one chimpanzee with hepatitis A virus (HAV) resulted in the replication of virus in liver, excretion of HAV particles in stool, and the appearance of circulating antibodies specific for hepatitis A. The development of an early antibody response in the chimpanzee and in one of the two infected marmosets was shown to interfere with the serologic detection of HAV antigen (HAV Ag) in homogenates of acute phase liver tissue obtained from these animals. Treatment of HAV Ag-positive and IgM anti-HAV-positive liver homogenates with thiol reducing compounds was shown to release HAV Ag from in vitro formed immune complexes. The increased RIA response for HAV Ag in homogenates treated with 2-mercaptoethanol (2-ME) or dithiothreitol (DTT) was further shown not to be due to activation of HAV Ag itself or to a nonspecific effect on the RIA coating antibody, radiolabeled probe, or homogenized liver tissue. IgG and IgM double-antibody sandwich RIAs for HAV Ag were also compared for their ability to detect HAV Ag under reducing and nonreducing conditions. Application of the 2-ME or DTT treatment procedure to the serologic detection of other viral antigens or viruses whose presence in blood, stool, tissue macerate, or other milieu may be masked by specific antibody appears to be feasible.
D000818:Animals; D000914:Antibodies, Viral / Q000276:immunology*; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral / Q000276:immunology*; D002143:Callitrichinae; D002499:Centrifugation, Density Gradient; D004229:Dithiothreitol / Q000494:pharmacology; D006506:Hepatitis A / Q000276:immunology*; D035922:Hepatitis A Antibodies; D006507:Hepatovirus / Q000276:immunology*; D008623:Mercaptoethanol / Q000494:pharmacology; D002679:Pan troglodytes; D011863:Radioimmunoassay
D000914:Antibodies, Viral; D000936:Antigen-Antibody Complex; D000956:Antigens, Viral; D035922:Hepatitis A Antibodies; D008623:Mercaptoethanol; D004229:Dithiothreitol