Separation of T and B lymphocytes by preparative cell electrophoresis

Free flow electrophoresis can separate mouse spleen and lymph node lymphocytes into two fractions: a high mobility (more negatively charged) and a low mobility (less negatively charged) fraction. The high mobility lymphocytes are sensitive to anti‐Θ serum and complement but resistant to an anti‐serum against mouse specific B lymphocyte antigen (MBLA). They produce rosettes with sheep red cells and induce a graft‐vs‐host reaction when injected into F 1 ‐hybrid recipients. The high mobility lymphocytes also respond to phytohemagglutinin in vitro and are cytotoxic to allogeneic target cells after appropriate immunization. The low mobility lymphocytes are sensitive to anti‐MBLA serum and complement but resistant to anti‐Θ serum, form rosettes and hemolytic plaques against sheep red cells after immunization, but neither respond to phytohemagglutinin in vitro nor induce a graft‐vs‐host reaction. In CBA mice the percentages of high and low mobility lymphocytes in the spleen and lymph nodes correspond to the known distributions of T and B cells in these organs. The separated cells are viable since they are able to perform biological functions. Thus free flow cell electrophoresis seems to produce viable populations of mouse T and B lymphocytes.