title: Comparison of immune responses induced in mice by vaccination with DNA vaccine constructs expressing mycobacterial antigen 85A and interleukin-21 and Bacillus Galmette-Guérin.
abstract: In this paper, we addressed the immune adjuvant effects of interleukin(IL)-21 on DNA vaccine constructs expressing mycobacterium tuberculosis (TB) Ag85A and compared immune responses induced in mice inoculated DNA vaccine constructs expressing Ag85A and IL-21 with mice inoculated DNA vaccine constructs expressing Ag85A alone or Bacillus Galmette-Guérin(BCG.). In this experiment, the gene of IL-21 was firstly amplified from plasmid pcDNA3.1-mIL21 by PCR and cloned into the plasmid pRSC, forming recombinant plasmid pRSC-IL21. Then, the gene of Ag85A was amplified from the plasmid pIRES-Ag85A by PCR and cloned into the recombinant pRSC-IL21 again, finally forming co-expression DNA vaccine constructs pRSC-IL21-Ag85A. It was identified by the analysis of endonuclease digestion, DNA sequencing, the IL-21 and Ag85A expression in SP2/0 cells. Mice were i.m. immunized with BCG, DNA vaccine constructs pRSC-Ag85A or pRSC-IL21-Ag85A respectively, and the immune responses induced in mice was compared with other vaccines. The results showed that the DNA vaccine constructs pRSC-IL21-Ag85A was successfully constructed since the Ag85A and IL-21 was correctly expressed in SP2/0 cells respectively, and it elicited stronger immune responses in Balb/c mice than that of mice immunized with pRSC-Ag85A and the efficiency was as BCG did. We concluded that the IL-21 was a promising immune adjunctive modality to enhance immunigenicity of DNA vaccine containing Ag85A and the study provided the possibility of further development of immune accessory effect of IL-21 on DNA vaccine against TB.
authors.affiliation: Department of Pathogenic Biology and Immunology, Southeast University School of Basic Medical Science, Nanjing, China. njdoujun@yahoo.com.cn
authors.forename: Jun
authors.initials: J
authors.lastname: Dou
authors.forename: Quan
authors.initials: Q
authors.lastname: Tang
authors.forename: Fengshu
authors.initials: F
authors.lastname: Zhao
authors.forename: Lili
authors.initials: L
authors.lastname: Chu
authors.forename: Junsong
authors.initials: J
authors.lastname: Chen
authors.forename: Minggang
authors.initials: M
authors.lastname: Cao
authors.forename: Chunsheng
authors.initials: C
authors.lastname: Liu
authors.forename: Yongfang
authors.initials: Y
authors.lastname: Wang
authors.forename: Yating
authors.initials: Y
authors.lastname: Li
authors.forename: Jin Long
authors.initials: JL
authors.lastname: Li
mesh_terms: D000276:Adjuvants, Immunologic / Q000235:genetics; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood / Q000276:immunology; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D007378:Interleukins / Q000235:genetics / Q000276:immunology*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D009163:Mycobacterium bovis / Q000276:immunology*; D010957:Plasmids / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D032581:Tuberculosis Vaccines / Q000096:biosynthesis / Q000276:immunology*; D019444:Vaccines, DNA / Q000096:biosynthesis / Q000276:immunology*; D000098546:Interleukin-21
chemical_list: D000276:Adjuvants, Immunologic; D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D007378:Interleukins; D032581:Tuberculosis Vaccines; D019444:Vaccines, DNA; D000098546:Interleukin-21