The Preliminary Study Of Cytokines Secretion Of Human's Different Immune Cells Induced By A New Peptide Mimic Of HCV HVR1

There has no effective treatment and prophylaxis way for chronic HCV infection now.HCV therapeutics vaccines, which can induce cellular immune response,can re-established immune response to HCV in chronic HCV infection patients and utilize self immune function to eliminate virus, so therapeutics vaccines may reduce virus relapse and become one of the important treatment in the future. The research was established on the study of HCV strains(lb/2a) envolpe protein E2 hypervariable regions 1(HVR1) sequence variation and synthesis a series of HCV HVR1 mimic according to the conservation site. The mechanism of inducing immue reaction by one of the mimics which was comprised with 7 amino acids (7P) had been demonstrated. On the baes of that, we wanted to investigate the action of 7P to the cytokine secretions of different immune cells of people. The study was devided to 3 sections.(1) the study of cytokines secretion of health people's different immune cells induced by 7PIn this section; we cultured the peripheral blood mononuclear cells (PBMCs) with 7P and then demonstrated the immune reaction by analyzing the cytokines secretion with intracellular cytokine staining. The cytokines included interferon-γ(IFN-γ), interleukin-10(IL-10), interleukin-4(IL-4) and tumor necrosis factor-α(TNF-α). We found that the immune reaction to the 7P which was represented by the cytokines secretion of health controls was homogeneous; in these actions there was some difference between the CD4~+CD8~-, CD4~-CD8~+ T cells and NK, NKT cells, and the change of cytokines secretion might contribute to the anti-virus action and lessen of inflammatory reaction. (2) the preliminary study of mechanism of 7P to the immune cellsIn this section, we first cultured the peripheral blood mononuclear cells (PBMCs) with 7P and then analysed the mature phenotypic and cytokines secretion of antigen-presenting cells, the results showed tha CD83 which was the mature phenotypic of DC lowered in mDC and had no significant changed in pDC, and did not found the secretion of IL-12 in mDC. Then we magnetically isolated antigen-presenting cells (APCs) from PBMCs with the Beads of MultiSort release and stop reagents. The remains contained T cells and NK, NKT cells were cultured with 7P to detect the cytokines secretion with intracellular cytokine staining again to determine the path of 7P action.. The results showed that when we magnetically isolated antigen-presenting cells (APCs) from PBMCs and cultured remains with 7P, we got the same immune reaction with that before isolation. Together, our findings suggested the action of 7P was maybe to the effector cell directly, and not through the antigen-presenting action of APCs. Meanwhile 7P maybe had some effect on the APCs, and this effect could influence the cytokines secretion of the effect cells directly or indirectly.(3) the study of cytokine secretion of HCV infected patients' different immune cells induced by 7PIn this section,we cultured the peripheral blood mononuclear cells (PBMCs)of HCV infected people with 7P and then demonstrated the immune reaction by analyzing the cytokines secretion with intracellular cytokine staining again, the result was partly same with and partly near to that of health controls which suggested 7P could change immune reaction through modulating the secretion of cytokines in HCV infected peoples too, and the change of cytokines secretion might contribute to the anti-virus action and lessen of inflammatory reaction.