PREPARATION OF MURINE TRANSPLANTATION ANTIGENS

Publisher Website
Google Scholar
Abstract
A simple method for the preparation of murine cell-free transplantation antigen utilizing homogenization of splenic tissue in buffered 0.25M sucrose solution is described. When the nuclear and cytoplasmic fractions of the splenic homogenate are separated, all of the antigenic activity, as assayed by accelerated skin graft rejection, resides in the cytoplasmic fraction. Ultracentrifuge analysis of the cytoplasmic fraction reveals that the antigen sediments over a wide range of gravitational field strengths. The major portion of the antigen in the cytoplasmic fraction is obtained in the supernatant remaining after the cytoplasmic fraction is centrifuged at 5000g (10 min) or in the sediment resulting from centrifuging this supernatant at 105,000g (60 min). The antigen so prepared is remarkably stable to freezing, lyophilization and storage at −20 C and to prolonged exposure to alkali (pH 9.5). The dose response of the antigen preparation as determined by the sensitizing capacity of decreasing amounts of antigen is approximately linear. Very small amounts of antigen cause significant sensitization. When compared with viable intact spleen cells, the antigen is as potent as the equivalent amount of cells from which it is extracted. Antigen in the form of the 5000g supernatant sensitizes equally as well by the i.v. as by the i.p. routes. Furthermore, the onset of sensitivity is rapid and its duration quite short by both routes. Repeated injections of sensitizing antigen by the i.v. or i.p. route prolong allograft survival, although prolongation is more easily achieved by the i.v. route.

This publication has 0 references indexed in Scilit: