Recombinant E. coli LLO/OVA induces murine BMDC maturation and chemokine and chemokine receptor expression
AIM: To investigate the effects of recombinant E. coli LLO/OVA on the maturation, the expression of chemokines and chemokine receptors of murine dendritic cells. METHODS: After bone marrow-derived dendritic cells (BMDCs) were pulsed by E. coli LLO/OVA and E. coli OVA respectively, the mRNA expressions of chemokines and chemokine receptors and NF-κB signaling pathway associated molecules were detected by supperar-ray hybridization and RT-PCR. The expressions of co-stimulatory molecules, MHC class Ⅱ and CXCR4 of these cells were determined by flow cytometry. RESULTS: The immature murine BMDCs expressed chemokine genes Ccl2, Ccl4, Ccl6, Ccl9, Ccl17, Ccl22, Cxcl2 and Cxcl4, and chemokine receptor genes Ccr2, Ccr5 and Ccr7. After stimulated by E. coli LLO/OVA for 4 -8 h, the BMDCs up-regulated the expression of genes Myd88, NF-κbl, Ccl3, Ccl5, Ccl7, Ccl22, Cxcl1, Cxcl9 and Ccr12, and expressed Ccr7 persistently, but down-regulated the expression of genes Ccr2 and Ccr5. After pulsed by E. coli LLO/OVA for 8 h, the BMDCs up-regulated the expression of genes Tlr2, Myd88, NF-Kb1 and Cxcr4. After pulsed by the vaccine for 12 h, the BMDCs down-regulated the expression of a series of chemokine genes, and only expressed Ccl5 and Cxcr4 persistently. The expressions of CD40, CD80, CD86, MHC class Ⅱ and CXCR4 were up-regulated after BMDCs were pulsed by E. coli LLO/OVA for 24 h. CONCLUSION: Recombinant E. coli LLO/OVA in- duces murine BMDCs maturation and up-regulates a series of cke-mokines and chemokine receptors genes, especially chemokine receptor CXCR4, probably by Myd88/NF-κB signaling pathway.
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