Preparation of monoclonal antibodies directed against the natural killer cell glycolipid asialo-gm1

Abstract

Natural killer (NK) cells are a subpopulation of large, granular lymphocytes that are able to lyse various types of tumor and virus infected cells without prior sensitization. Because of their tumor surveillance characteristics, NK cells have been subjected to many studies in humans and mouse systems. One problem with studying NK cells is that they have shared surface molecules with other cells which makes it difficult to identify them. Several years ago it was discovered that mouse NK cells express the cell surface molecule ganglio-N-tetrasylceramide (asialo-GM1). The molecule is pressed primarily on NK cells and is thought to play a role in their cytolytic activities. Currently, commercially available polyclonal antibodies against asialo-GM1 are used in vivo and in vitro studies to identify NK cells. Unfortunately these antibodies are nonspecific and thus cross react with other molecules. To bypass this problem with utilized the hybridoma technology to make monoclonal antibodies directed against asialo-GM1 which should be more specific and less cross reactive. In this study we immunized NZW mince with asialo-GM1 and fused its spleen cells with a tumor cell to make a hybrid cells. From these hybrids, five clones which are secreting antibody against asialo-GM1 were isolated. The products of these hybrid will be used as a research tool for further studies on natural killer cells.