Fig. 8. Effect of N-isobutyl-dodecatetraenamide on cytokine production in CD4+ T cells under Th2-skewing condition. Bone marrow cells were obtained from the femur and tibia of naïve mice. Dendritic cells (DC) were prepared for co-culture by performing CD11c MACS enrichment in bone marrow derived dendritic cells. CD4+ T cells were enriched by collecting and pooling spleens and peripheral lymph nodes from 6-week male C57BL/6N mice, followed by CD4 MACS enrichment. The enriched samples were identified and flow-sorted for naïve CD4+ gated as CD4+CD44lowCD25-CD62Lhi using FACSAria III cell sorter. For DC - CD4+ T cells co-culture assays, cells were co-cultured in complete media with soluble anti-CD3ε Ab (clone 145-2C11), anti-IFNγ (clone XMG1.2), IL-2, IL-4 in the presence of N-isobutyl-dodecatetraenamide for 3 days. Intracellular cytokines were analyzed in CD4+ T cells under Th2-skewing condition targeting (A) IL-4 and (B) IL-13. From the CD4+ T cells co-cultured with dendritic cells, (C) IL-13 production was analyzed. All the data are expressed as the mean ± SEM (n=5 per group). *, **, *** Significantly different (p<0.05, 0.01, 0.001) from Control group.
© Biomolecules & Therapeutics
