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Production of Precipitating Primate Antibodies to Human Membrane Antigen(s) with HL-A Activity by Immunization with Soluble HL-A Antigens¹

From the Departments of Microbiology and Immunology, Duke University Medical Center, Durham, North Carolina and Yerkes Primate Research Center, Emory University, Atlanta, Georgia

Abstract

A monkey immunized with papain-solubilized HL-A antigens which had been purified on a Sephadex G-150 column produced antibodies which gave a preciptation reaction with the immunizing material as well as with certain other papain soluble spleen antigen preparations. The precipitating antibodies could not be removed by absorption with human serum but could be removed by absorption with peripheral blood leukocytes from all human donors tested. Purified suspensions of blood lymphocytes, lymph node cells or thymocytes were unable to absorb the precipitating antibody. Supernatants of saline suspensions of buffy coat leukocytes "released" antigen which precipitated with the monkey antiserum and gave reactions of identity by agar precipitation with the immunizing papain-solubilized antigen. Peripheral blood leukocytes from a patient with a leukemoid reaction containing 98% neutrophils were able to release precipitating antigen whereas purified blood and lymph node lymphocytes and thymocytes were not. The papain-solubilized antigen preparations as well as the released antigen preparations contained specific HL-A alloantigenic activity as well as precipitating antigen. Precipitation of the papain soluble antigen preparations as well as released antigen by the monkey antiserum resulted in a loss of the HL-A activity from the supernatant which could be recovered in the immune precipitate. These findings suggest that the monkey precipitating antibodies may be reacting with the HL-A base molecule(s) since they do not appear to interfere or react with site(s) conferring HL-A alloantigenic specificity.

Footnotes

¹ This work was supported by National Institutes of Health Grants AM-08054 and CA-08975 to Duke University and FR-00165 to Yerkes Primate Research Center.

² Present address: Naval Medical Research Institute, Bethesda, Maryland 20014.