An In Vitro Study On The Immunological Effects Of IFN-inducible T Cell Alpha Chemoattractantin Craft Rejection

Objective The present study aims to investigate the expression and immune functions of IFN-inducible T cell alpha chemoattractant (I-TAC), a member of the CXC chemokine family, and the inhibitory effect of mycophenolate acid (MPA) on I-TAC, thereby to observe the role of I-TAC in organ transplantation and the possible mechanisms of MPA in clinical therapy for allograft rejection.Methods The expression of I-TAC mRNA in human umbilical vein endothelial cell line ECV304 was analyzed by RT-PCR. The culture supernatants from mixed human peripheral blood mononuclear cells (PBMC) were quantitated for IFN-γand TNF-αproduction by ELISA. The PBMC migration attracted by I-TAC was determined by the use of chemotaxis chamber. The promotive capacity of I-TAC in PBMC adhesion to ECV304 cells was evaluated by adhesion assay. The effect of I-TAC on the proliferation of PBMC and ECV304 cells was detected by MTT assay. The expression of the I-TAC receptor, CXCR3, on PMBC and ECV304 cells was measured by Flow cytometry (FCM).Results Incubation of ECV304 cells with IFN-γfor 8 hours resulted in the expression of I-TAC mRNA. The addition of TNF-αincreased this inductive capacity of IFN-γin a dose-dependent manner. Significant production of IFN-γand TNF-αwas detected from the mixed culture of the PBMC from two individuals, which supernatants induced the expression of I-TAC mRNA in ECV304 cells. In the PBMC migration assay, the migration index markedly increased in I-TAC treated groups, and reached the peak value at the I-TAC concentration of 200ng/ml. I-TAC significantly enhanced the proliferation of PBMC and their adhesion to ECV304 cells, but inhibited the proliferation of ECV304 cells. I-TAC receptor CXCR3 molecules were detected on the surfaces of ECV304 cells and the mixed PBMC cells from two individuals.When MPA was applied to the incubation setups described above, IFN induced I-TAC...