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In vitro detection of lymphokines produced by in vivo activated lymphocytes
1Department of Immunology, Lilly Research Laboratories, Corporate Center, Indianapolis, IN 46285.
The purpose of these experiments was to develop a method to measure the production of lymphokines by cells which were activated by antigen in vivo. Previous protocols have been relatively unsuccessful since small, if any, amounts of lymphokines were available for measurement when in vivo antigen activated lymphocytes (AAL) were examined. These unsuccessful experiments usually employed supernatants derived from in vivo AAL which had been cultured in vitro for 4-24 h. As an alternative to assaying supernatants for the presence/absence of lymphokines, we have developed a co-culture system in which the indicator cells are directly added to the wells containing the in vivo AAL. Utilizing this system in conjunction with appropriate neutralizing monoclonal antibodies, we have demonstrated that interleukin-2, interleukin-3/colony stimulating factor and tumor necrosis factor-alpha can be readily detected from in vivo AAL.
The purpose of these experiments was to develop a method to measure the production of lymphokines by cells which were activated by antigen in vivo. Previous protocols have been relatively unsuccessful since small, if any, amounts of lymphokines were available for measurement when in vivo antigen activated lymphocytes (AAL) were examined. These unsuccessful experiments usually employed supernatants derived from in vivo AAL which had been cultured in vitro for 4-24 h. As an alternative to assaying supernatants for the presence/absence of lymphokines, we have developed a co-culture system in which the indicator cells are directly added to the wells containing the in vivo AAL. Utilizing this system in conjunction with appropriate neutralizing monoclonal antibodies, we have demonstrated that interleukin-2, interleukin-3/colony stimulating factor and tumor necrosis factor-alpha can be readily detected from in vivo AAL.