Al (oh) 3 And Cpg Odn Immunoadjuvant Multi-epitope Antigen Pcx3 Immune Effects Of The Impact Of The Hepatitis C Virus

HCV is a positive RNA virus,which genome is approximately 9.6 kb and encodes a polyprotein of 3010 to 3030 aa residues.From N-to C-terminus of the polyprotein,the component proteins are core protein(C),envelope glycoproteins(E1, E2),p7,and non-structural proteins NS2,NS3,NS4A,NS4B,NS5A and NS5B.The hight rate of mutation makes virus easy to escape from the host defenses,which is one of the major obstacle in the vaccine development.Recent study indicated that the effective vaccine must elicit the high-level,broad(refer to squasis) and specific immune response.A multi-epitope antigen gene pox was assembled by five highly conserved B-and /or T-cell epitopes,which were selected from C(1～16 aa,132～141 aa),E1 (126～135 aa),NS3(139～147 aa) and NS5B(768～775 aa) protein,as well as a T-cell epitope of tetanus toxin(TT),which was added to enhance T-cell immunity. The pcx3 gene was constructed by triple tandem of pcx gene as designed and was further cloned into pET-28b(+).PCX3 protein was expressed in E.coli purified up to 98%by Ni²⁺-NTA-agarose.The antigen-HCV protein vaccine PCX3 with Al(OH)₃ adjuvant,CpG ODN adjuvant,and the combination of them were separately used to immunize six-week-old male BALB/c mice.Compared with other groups,the group which has PCX3 antigen and the combined-adjuvant of Al(OH)₃ and CpG ODN,was detected the highest specific IgG(1×10⁶) and IgM(1×10⁵) antibody titer and the longest lasting time by ELISA method;When detected the natural killing effect of NK cells (CTL) with a non-radioactive cytotoxicity assay kit,the CTL reponse(40.91%) significantly higher than other groups(P＜0.05);And when analyzed lymphocytes subsets of whole blood by flow cytometry technology,since the 2th week,the ratio of CD4⁺/CD8⁺ T lymphocyte was significantly higher than the group with PCX3 antigen alone(P＜0.05).Conclusion:the combined-adjuvants of Al(OH)₃ and CpG ODN can enhance the immune effect of HCV protein vaccine PCX3 in both humoral immunity and cellular immunity aspect,and has a synergistic effect.