A SIMPLE AND SENSITIVE METHOD TO STUDY EFFECTS MEDIATED BY SOLUBLE LYMPHOKINES AS DEMONSTRATED BY THE INTERACTION OF CD4-CELL ACTIVATION( AND CD8+ CELL SUBSETS DURING T)

A method is described for the study of lymphokine-mediated cellular in teractions using triple wells, which permits co-culture of cell subpop ulations without direct physical contact. The triple wells are constru cted by slitting the walls to half height between three adjacent wells of a 96-well microtiter plate. The cells under study are positioned i n the outer two wells, whereas the middle well serves to separate the cells. The half slits permit the wells to be treated independently bef ore filling the triple well with the culture medium and prevents cell leakage thereafter. The feasibility of the method was established by s tudying the interaction of isolated CD4(+) and CD8(+) T cell subsets d uring T cell proliferation induced by immobilized anti-CD3 and anti-CD 28 monoclonal antibodies.