Effects of the endogenous CRF on the biological phenotypes of mouse mesenteric lymph nodes den-dritic cells

Abstract

Objective To investigate the expression of corticotropin-releasing factor (CRF) and its receptors including CRFR1 and CRFR2 on mouse mesenteric lymph nodes dendritic cells (MLNDC), and to analyze their effects on the biological phenotypes of intestinal dendritic cells. Methods The MLNDCs were isolated from C57BL/6 mice by using magnetic bead sorting. The purity of CD11c+ DCs was identified by flow cytometry. The double-labeling immunofluorescence and the reverse transcriptase-polymerase chain reaction (RT-PCR) were performed to detect the expression of CRF, CRFR1 and CRFR2 on MLNDCs. The MLNDCs were exposed to CRF with or without the interference of CRFR1 and CRFR2 antagonists. Flow cytometry was used to measure the changes of surface molecules (MHCⅠ and MHCⅡ) and co-stimulatory molecules (CD80 and CD86). Results The CD11c+ DCs accounted for (80.12±6.34)% of the isolate cells with a high cell viability of more than 90%. The expression of CRF, CRFR1 and CRFR2 at mRNA level were detected in MLNDCs by RT-PCR. Results of the immunofluorescent staining assay indicated that both CRFR1 and CRFR2 were expressed on the surface of MLNDCs. The expression of CD86 on MLNDCs was inhibited by the treatment of MLNDCs with CRFR1 antagonist, but enhanced by the treatment with CRFR2 antagonist. Conclusion Both CRF and CRFRs were detected in the MLNDCs isolated from the C57BL/6 mice. The CRF could alter the biological phenotypes of MLNDCs through binding to different CRFRs (CRFR1 and CRFR2), which affected the phenotypes of MLNDCs in opposite ways. Key words: Corticotropin-releasing factor; Mesenteric lymph nodes; Dendritic cell