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[Experimental study of immunization of mice with hepatitis C virus genetic vaccine constructs]
1Institute of Infectious Disease, The First Affiliated Hospital of Zhejiang University, Hangzhou 310003.
Objective:
To inquire into the immune responses to expression protein in mice immunized with genetic vaccine of hepatitis C virus (HCV) and lay a foundation for HCV genetic vaccine development in future.
Methods:
The gene fragments coding C and most E regions of HCV-II type were inserted into pCD-SRalpha(1) of eukaryotic expression vector and formed genetic vaccine constructs of pCD-HCV(1) and then was injected into the quadriceps muscles of Balb/c mice. The serum anti-HCV level of mice was tested by ELISA and peripheral blood mononuclear cell (PBMC) proliferative responses to HCV antigens were detected by (3)H-TdR incorporation method (cpm).
Results:
The serum antibody level reached to 0.71 +/- 0.08 - 0.77 +/- 0.06 (A value, the same below) after genetic vaccine pCD-HCV(1) (100 microg/mouse) were inoculated into the mice (n = 12) three or four times while blank vector pCD-SRalpha(1) could not induce the mice (n = 8) to generate antibody response in same way. After the antibody levels in mice (n = 8) immunized by pCD-HCV(1) had ascended to peak value (0.71), there was no trend of descending during the following 18 weeks of detection (0.68 +/- 0.06 - 0.75 +/- 0.07). Specific fragment of HCV cDNA identified by polymerase chain reaction (PCR) from DNA extracted from the muscles of the mice after pCD-HCV(1) had been inoculated three months. PBMC proliferative responses to HCV synthetic peptides CP(9) and gene recombinant antigens C, E(1) in the mice immunized with pCD-HCV(1) were detected and its stimulation indexes (SI) were 4.07 +/- 1.58, 3.88 +/- 0.70 and 3.69 +/- 1.13 respectively and there was a significant difference (P < 0.001) as compared with that of PBMC in mice immunized with pCD-SRalpha(1).
Conclusion:
These investigations demonstrated that genetic vaccine constructs made of HCV structural region can induce Balb/c mice to generate antibody and PBMC proliferative responses to HCV antigens via DNA immunization.
