Immunogenicity of mutant and wild HPV16 DNA vaccines.
- Author:
Ya-gang ZUO
1
;
Jia-bi WANG
;
Hong-zhong JIN
;
Liu YUE-HUA
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Enzyme-Linked Immunosorbent Assay; Interferon-gamma; blood; Interleukin-2; blood; Male; Mice; Mice, Inbred C57BL; Mutation; Oncogene Proteins, Viral; genetics; immunology; Papillomaviridae; genetics; immunology; Papillomavirus E7 Proteins; Recombinant Fusion Proteins; immunology; T-Lymphocytes, Cytotoxic; immunology; Vaccination; Vaccines, DNA; immunology; Viral Proteins; biosynthesis; genetics; Viral Vaccines; immunology
- From: Acta Academiae Medicinae Sinicae 2004;26(5):554-557
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo evaluate the efficacy of mutation of human papillomavirus 16 (HPV16) E7 in two zinc-binding motifs on HPV16 E7 C terminus on antigen-specific immunity.
METHODSpcDNA3.1/16E7 and pcDNA3.1/ME7 were successfully constructed by inserting the E7 (ME7) into pcDNA3.1 BamH I, EcoR I cut sites. After intramuscular injection with pcDNA3.1, pcDNA3.1/16E7, and pcDNA3.1/16ME7 on C57BL/6 mice, splenocytes from vaccinated mice was isolated. After have been stimulated with E7-specific peptide, interferon gamma (IFN-gamma), interleukin 2 (IL-2), and cytotoxic T lymphocyte (CTL) were detected by ELISA, Eli-spot, and LDH assay respectively; splenocytes without E7 peptide stimulation were used as control group.
RESULTSSplenocytes from mice vaccinated with pcDNA/ME7, stimulated with E7 peptide, generated significantly larger number of E7-specific IL-2 compared with pcDNA3.1/16E7, pcDNA3.1, and control group. The E7-specific IL-2 generated in pcDNA-ME7 group was 5-fold of that of pcDNA3.1/16E7, and the difference was statistically significant (P < 0.05). Splenocytes from mice vaccinated with pcDNA/ME7 and stimulated with E7 peptide, generated significantly larger number of E7-specific IFN-gamma compared with other vaccines. pcDNA-ME7 generated a 2-fold increase in the number of E7-specific IFN-gamma compared with wild-type E7 and the difference was statistically significant (P < 0.05). The highest CTL activity in mice vaccinated with pcDNA/ME7 at an E:T ratio of 45:1 was achieved compared with mice vaccinated with other vaccines. The percents of specific lysis generated by pcDNA3.1/ME7, pcDNA3.1/E7, pcDNA3.1, and without vaccination were of (28.7+/-1.2) %, (55+/-2.2) %, (12.5+/-2.0) %, and (11.5+/-1.2) % respectively, and significant difference existed between the former and the latter two groups (P < 0.05). However, no significant difference was found among all the groups without specific E7 peptide stimulation (P > 0.05).
CONCLUSIONSThe mutation of zinc-binding motifs on HPV16 E7 C terminus may greatly enhance the immunogenicity.
