1.Impact of compound 48/80 activated mast cell on dendritic cell homing to lymph nodes
Shurong REN ; Qiubo WANG ; Yanli ZHANG ; Cuixiu LU ; Wenli LI ; Wenjuan HU
Chinese Journal of Immunology 2017;33(8):1146-1151
Objective:To investigate the effect of compound 48/80(C48/80) activated mast cell on dendritic cell homing to draining lymph nodes(DLN) in order to explore the potential mechanism of C48/80 promoting adaptive immune response.Methods: C57BL/6 mice bone marrow derived dendritic cells(BM-DC) and MC/9 mast cell line were propagated in vitro.Chemotaxis of BM-DC supplemented with supernatant of C48/80 treated MC/9 to CCL21 was measured by transwell chemotaxis assay.C48/80 was injected into murine scapular site,local skin mast cell degranulation was detected by toluidine blue staining method.The number of CD11c+ DC in draining lymph nodes(DLN) was detected by Flow cytometry.Exogenous green microbeads labeled BM-DC homing to DLN was detected by fluromicroscope.The mice pretreated with or without C48/80 were vaccinated by ovalbumin pulsed DC(OVA-DC),DLN lymphocyte proliferation was detected by WST-8 reagent.Results: A large amount of typical BM-DC were harvested from media supplemented with murine recombinant IL-4 and GM-CSF.Product of activated MC/9 enhanced chemotaxis of BM-DC to CCL21(P<0.01).Intradermal injection of C48/80 induced local mast cell obvious degranulation and local inflammation.Mice pretreated with C48/80 demonstrated high number of total cells and DC cells in DLN.The number of fluorescent positive BM-DC in DLN also increased in C48/80 pretreated mice.Antigen specific lymphocyte proliferation enhanced in OVA-DC inoculated C48/80 pretreated mice.Conclusion: Mast cell activation induced by C48/80 can enhance DC homing to DLN and increase specific lymphocyte proliferation,which may be one of a mechanism of C48/80 in promoting adaptive immune response.