The new approach to produce monoclonal antibody toward differntiated antigens by the aid of immunotolerance.

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Recent studies have shown that the oligosacchrides in glycoproteins and glycolipids on the cell surface have inportant biological roles, such as cell adhesion and developmental events. Inview of the potential importance of ologosaccharide chain, i have tried to produce monoclonal antibodies which can discriminate differentiated antigens from commn antigens. By the aid of immunotolerance the most of the lymphoid cells which produce antibodies against the common antigen are eliminated from the immunized animals before injection of differentiated cells. It make us possible to produce specfic miniclonal antibodies toward differentiated antigens.Pheochromocytoma cells(PC12) and PC12D cells, a new subline derived from PC12, were used as a model antigen for this experiment. I have succeeded in the induction of immunotolerance by the elimination of helper T-cells with co-injection of cyclophosphamide, but failed to produce monoclonal antibodies recognized differntiated antigens. However, during the courceof this investigation I found that the amount of membrane glycoproteins deffered significantly between PC12 and PC12D cells. In the NGF induced differntiation of PC12 cells also led to significant decrease in the proportion of membrane glycoproteins which have polylactosamine chains on its molecules. These results suggest that there may be some correlation between the decrease in content of poly(N-acetyllactosaminyl) residues in goycoproteins and the changes in cell morphology.