Use and functional properties of peripheral blood lymphocytes in mice

Peripheral blood lymphocytes (PBL) were isolated and examined for their ability to respond in vitro to H-2 and H-Y antigens and also to hapten conjugated syngeneic cells. In addition, these lymphocytes were tested as antigen and target cells in in vitro mixed lymphocyte reaction and in chromium release cell mediated lympholysis respectively, as well as serving as targets for the H-2 typing of chimeric mice. Two methods were used to isolate the lymphocytes: a density gradient separation and a double water lysis technique. PBL, prepared by either method compared favourably with splenic lymphocytes in all aspects of anti-H-2 cytotoxicity but could not be used as responder for anti-H-Y cytotoxicity. In addition a method has been developed using PBL as targets for H-2 typing of both allophenic and irradiation chimeric mice. The small numbers of cells required for each of these procedures allows for the preselection of suitable mice and/or multiple experimental determinations on individual mice.