Self-priming cell culture system for monitoring genetically-controlled spontaneous cytokine-producing ability in mice

To monitor genetically-controlled cytokine-producing ability in mice in vit ro, we developed a high-density cell culture system, which is preferable fo r inducing CD4(+) T cell-dependent self-priming responses without any antig enic stimulation. When BALB/c spleen cells were cultured at high density (o ver 1.0 x 10(7) cells/well) in 12-well culture plate, they spontaneously pr oduced cytokines including IFN-gamma, IL-2, IL-3, IL-5 and IL-6. The sponta neous cytokine production in this self-priming cell culture (SPCC) system w as totally dependent on MHC class II-restricted CD4(+) T cells It was demon strated that Th2-type BALB/c background mice exhibited higher levels of spo ntaneous cytokine production in SPCC culture compared with Th1-type C57BL/6 mice. Moreover, using BALB/c x C57BL/6 F-1 mice and B10D2 congenic mice, i t was demonstrated that highly spontaneous cytokine-producing ability in BA LB/c background is genetically dominant and it is controlled by non-MHC gen es. Unexpectedly, BALB/c mice spontaneously produced higher levels of IL-2 and IFN-gamma than C57BL/6 mice. However, BALB/c mice revealed lower levels of CTL and NK cell-generation in SPCC system compared with C57BL/6 mice. T hese results suggested that genetically-controlled predisposition of BALB/c mice toward Th2 immunity appeared not to be derived from their poor IFN-ga mma-producing ability but rather derived from their poor responsiveness to IFN-gamma. (C) 1999 Elsevier Science B.V. All rights reserved.