A simple and sensitive method to study effects mediated by soluble lymphokines as demonstrated by the interaction of CD4+ and CD8+ cell subsets during T cell activation

Abstract A method is described for the study of lymphokine-mediated cellular interactions using triple wells, which permits co-culture of cell subpopulations without direct physical contact. The triple wells are constructed by slitting the walls to half height between three adjacent wells of a 96-well microtiter plate. The cells under study are positioned in the outer two wells, whereas the middle well serves to separate the cells. The half slits permit the wells to be treated independently before filling the triple well with the culture medium and prevents cell leakage thereafter. The feasibility of the method was established by studying the interaction of isolated CD4 + and CD8 + T cell subsets during T cell proliferation induced by immobilized anti-CD3 and anti-CD28 monoclonal antibodies.