Development of hepatitis virus C therapeutic vaccine with hepatitis virus B core antigen us the carrier
- VernacularTitle:乙型肝炎核心抗原为载体进行丙型肝炎混合性治疗疫苗的实验
- Author:
Jia-Yu CHEN
;
Fan LI
;
- Publication Type:Journal Article
- Keywords:
Hepatitis C;
Vaccines;
Epitopes,T-lymphocytes;
Hepatitis B core antigen
- From:
Chinese Journal of Infectious Diseases
2001;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To invesgate hepatitis B c antigen(HBeAg)as the carrier to construct mixed hepatitis C virus(HCV)therapeutic vaccine.Methods Fused the pTrc-core gene with two synthetic T-epitope antigen gene of HCV,expressed the plasmids pTrc-core-T1 and pTrc-core-T2, applied sucrose density gradient centrifugation to get the fusion protein HBcAg-T1 and HBcAg-T2, dialysised and concentrated the protein,mixed and immunized them in mice using the protein HBcAg (expressed by pTrc-core)as control.The tumor regression trial in mice was evaluated at appropriate time.After immunized four times,got the blood and spleen of mice.Interleukin(IL)-12 in serum were detected by enzyme-linked immunosorbent assay(ELISA),nonspecific T lymphocytes prolifera- tion response of splenic lymphocytes was respectively examined by thiazolyl blue(MTT)assay.T cell subset of splenic lymphoeytes,IL-5 in serum,IL-4 and interferon(IFN)-?in lymphocyte were evalu- ated by FACS.Results Tumor regression trial showed the experimental group formed only one tumor(diameter=0.1 cm),smaller than the T_1T_2 peptides group(diameter=0.9 cm)and blank group(diameter=1.3 cm).FACS indicated that CD8~+ T cell percentage of spleen cells from HBcAg- T_1T_2 group(20.21?2.01)% was higher than T_1T_2 peptides group(15.33?1.45)% and blank group(5.09?1.66)%,the percentage of IFN-?positive cells in these three groups were(1.58?0.05)%,(0.88?0.02)% and(0.53?0.03)%.The ELISA discovered that the level of IL-12 in the experimental group was the highest.Different from above,the IL-4 and IL-5 were lower in the exper- imental group.The detection of eytotoxic T lymphocyte(CTL)activity showed that the quantity of Hela cells infected by HCV in HBcAg-T1T2 stimulated group was different obviously from T1T2 peptides group. Conclusion The mixed protein HBcAg-T1 and HBcAg-T2 can induce stronger cellular immunity and it is able to serve as a therapeutic vaccines candidate specific for HCV.
