[Expression of cytokines in murine BMDCs induced by recombinant E.coli LLO/OVA via TLR4 and NOD1 receptors].

Aim: To investigate the relationship between the activation of pattern recognition receptors and the cytokines expression of dendritic cells.

Methods: After bone marrow-derived dendritic cells (BMDCs) were pulsed by E.coli LLO/OVA, the mRNA expression of pattern recognition receptors and the downstream NF-kappaB signal pathway associated molecules were detected by microarray hybridization and RT-PCR; the expression of costimulatory molecules, MHC class II and cytokines were determined by flow cytometry and ELISA.

Results: After BMDCs were pulsed by E.coli LLO/OVA for 2 h, the levels of TLR4, Myd88, Rip2, Irak1, Irak2, Ikkalpha, NF-kappaB1 and NF-kappaB2 mRNA up-regulated; and the levels of Icam1, IL-1a, IL-1b, IL-6 and TNF-alpha mRNA up-regulated also; after BMDCs were pulsed by E.coli LLO/OVA for 4 h, the levels of Card4(Nod1), Rip2, Ikkbeta, NF-kappaB1 and NF-kappaB2 mRNA up-regulated, meanwhile, the levels of TNF-gamma, TNF-beta and CD40 mRNA up-regulated. The expressions of costimulatory molecules and MHC class II of the BMDCs up-regulated and the concentration of IL-12 and IFN-gamma increased in the supernatant of BMDCs pulsed by E.coli LLO/OVA at 24 h.

Conclusion: Recombinant E.coli LLO/OVA induces maturation and expression of cytokines, especially IL-12 and IFN-gamma, of murine BMDCs through activation of NF-kappaB signal pathway with TLR4 and NOD1 receptors.