Authors: Elida Cleyse Gomes da Mata, Kanzaki L.I.B
The immunological response to any antigen is identified and quantified “in vivo” but the tools to perform “in vitro” the same procedures in cell culture are cumbersome and inaccurate. Usually lymphocytes are isolated by centrifugation in sucrose density gradient and cultured in RPMI medium requiring stimulation with phytohemagglutinin and maintained by the addition of interleukin-2 in the medium. Besides, “in vitro” assays to evaluate any natural or synthetic compound could be masked by the mitogen and IL-2 activity. An easy and practical approach to culture human leucocytes and evaluate the response to any natural compound or the immunological response to any pathogen is described. Instead of just lymphocyte isolation, all leucocytes were collected after separation from total peripheral blood cells in sucrose density gradient. Leucocytes in culture, representing different cell phenotypes, mimic “in vitro” cell cooperation during host exposition to foreign antigens. Therefore, the procedure described utilizes the filtered supernatant of lymphoblastic cell lines, HUT-78, to activate primary human leucocytes to divide and differentiate, maintained indefinitely under culture. In this way “in vitro” stimulated leucocytes with any immunogenic compound respond producing the corresponding cytokines, chemokines and antibodies. Among other
applications, viral production could be achieved including viral isolation. A large array of potential diagnostic and therapeutic applications could be devised utilizing this simple and new method.