Characterization of Human Lymphocyte Surface Receptors for Mitogenic and Non‐mitogenic Substances

Publisher Website
Google Scholar
Abstract
To compare the receptor patterns for mitogenic and non-mitogenic substances, surface glycoproteins of human lymphocytes were labeled with the lactoperioxidase-catalyzed iodination technique and with a galactose oxidase-tritiated sodium borohydride technique. Labeled cells were detergent-solubilized; the lysates were allowed to react with insolubilized purified mitogenic lectins, phytohemagglutinin, leukoagglutinin and an insolubilized non-mitogenic lectin, oxidized leucoagglutinin. Lectin-reactive proteins were eluted with sodium dodecyl sulphate (SDS) buffer. Cell membrane components reactive with anti-lymphocyte globulin (ALG) were retrieved by indirect immunoprecipitation with protein-A-bearing Staphylococcus Cowan I strain (SaCI). Lectin- and ALG-reactive proteins were analyzed by SDS polyacrylamide gel electrophoresis. Iodinated glycoproteins regularly showed 4 major components with MW of 120,000, 70,000, 60,000 and 43,000 daltons, respectively, on 7% gels. An additional broad peak in the MW range 20,000-35,000 daltons was on 10% gels. Tritiated glycoproteins showed 4 major components with MW 120,000, 70,000, 60,000 and 42,000, respectively, which reacted with lectin and ALG. In addition, ALG reacted with some glycoproteins with MW between 150,000 and 230,000 daltons. On 10% gels additional lectin- and ALG-binding glycoproteins with MW around 30,000 daltons were found. The similarity in structures bound by mitogenic and non-mitogenic substances indicates that lymphocyte activation may depend on some property conferred by the mitogen.