TFPR1 acts as a novel adjuvant by activating dendritic cells and promoting its matu-ration / 军事医学

ABSTRACT

Objective To study the role of dendritic cells in the function of a recombinant protein TFPR 1 as an adjuvant .Methods Bone marrow cells were collected from four-to five-week-old male BALB/c mice under aseptic conditions, and cultured with complete RPMI 1640 containing rmGM-CSF and rmIL-4 for six days.TFPR1 was added on day 6, and cells were incubated for another 24 hours.LPS was used as positive control , while PBS as negative .The morphology of dendritic cells was observed under an optical microscope and laser confocal microscope , cell surface makers (CD40,CD80,CD86 and MHCⅡ)were detected with flow cytometry, and the cytokines in the supernatant were detected with ELISA.Results Compared with negative control ,dendritic cells incubated with TFPR1 for 24 hours were significantly different in morphology as was observed by optical and laser confocal microscopes , but were similar to positive control .Most of the dendritic cells treated with TFPR 1 showed less adherence and became round , whose podosomes became shorter , and even disappeared .Actin distribution changed from two poles of the cell to the membrane .CD40,CD80, CD86 and MHCⅡon the cell surface were up-regulated on stimulation by TFPR1,as was detected by FACS.These results showed that TFPR1 was capable of promoting dendritic cell maturation .ELISA showed dendritic cells treated with TFPR 1 secreted high levels of cytokines(IL-6, IL-8 and TNF-α).Conclusion TFPR1 is capable of promoting dendritic cell maturation , and activating cells to produce cytokines , indicating that dendritic cells can play an important role in the function of TFPR 1 as a novel ad-juvant .