title: Immunogenicity and protective efficacy of recombinant Bacille Calmette-Guerin strains expressing mycobacterium antigens Ag85A, CFP10, ESAT-6, GM-CSF and IL-12p70.
abstract: This study aimed to evaluate the immunogenicity and protective efficacy of recombinant bacille calmette-guerin (rBCG) strains expressing Ag85A (A), CFP10 (C), ESAT6 (E), IL-12p70 (I), and fusion protein GM-CSF (G).
  
  
  
  rBCGs were established by integrating of A, C, E, I, G, AE, CE, IE, GC, GE and GCE into Mycobacterium bovis BCG-1173 and BCG-SH. The macro-effects of rBCGs on mice were evaluated by phenotype and weight. The immunogenicity of rBCGs was analyzed by lgG, lgG1 and lgG2a antibody titers, and IFN-γ and IL-4 contents through Enzyme-linked immunosorbent assay (ELISA). Meanwhile, the proportions of CD4+and CD8+T splenic lymphocytes were determined using flow cytometry. The protective efficacy of rBCGs was evaluated by bacterial load in spleen and lung tissues from immunized mice.
  
  
  
  rBCGs exhibited no obvious side effects on mice. The antibody titers of lgG, lgG1 and lgG2a, proportion of CD4+and CD8+T cells, and concentrations of IFN-γ were found to be significantly higher in multiple-gene rBCGs than that in single-gene rBCGs (P < 0.05). Bacterial load in both spleen and lung tissues from mice infected with M. tuberculosis H37Rv were significantly reduced by rBCGs. A significantly lower bacterial load was revealed in rBCG-1173:A compared with multiple-gene rBCGs (P < 0.05).
  
  
  
  Immunogenicity was better on multiple-gene rBCGs than on single-gene rBCGs, while excellent protective efficacy was exhibited on rBCG-1173:A and BCG-1173.
authors.affiliation: a Laboratory of Infection and Immunity, West China Center of Medical Science , Sichuan University , Chengdu , China.|b Department of Pathogenic Biology , School of Basic Medical Science, Chengdu Medical College , Chengdu , China.
authors.forename: Fu-Ying
authors.initials: FY
authors.lastname: Dai
authors.affiliation: a Laboratory of Infection and Immunity, West China Center of Medical Science , Sichuan University , Chengdu , China.
authors.forename: Jun-Fang
authors.initials: JF
authors.lastname: Wang
authors.affiliation: a Laboratory of Infection and Immunity, West China Center of Medical Science , Sichuan University , Chengdu , China.
authors.forename: Xue-Li
authors.initials: XL
authors.lastname: Gong
authors.affiliation: a Laboratory of Infection and Immunity, West China Center of Medical Science , Sichuan University , Chengdu , China.
authors.forename: Lang
authors.initials: L
authors.lastname: Bao
mesh_terms: D000217:Acyltransferases / Q000235:genetics / Q000276:immunology*; D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D000942:Antigens, Bacterial / Q000235:genetics / Q000276:immunology*; D001500:BCG Vaccine / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D001426:Bacterial Proteins / Q000235:genetics / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology; D004195:Disease Models, Animal; D005260:Female; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics / Q000494:pharmacology*; D007371:Interferon-gamma / Q000378:metabolism; D018664:Interleukin-12 / Q000235:genetics / Q000494:pharmacology*; D015847:Interleukin-4 / Q000378:metabolism; D008168:Lung / Q000382:microbiology; D008807:Mice, Inbred BALB C; D009163:Mycobacterium bovis / Q000235:genetics / Q000276:immunology; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000276:immunology; D013154:Spleen / Q000382:microbiology; D014376:Tuberculosis / Q000517:prevention & control*; D014614:Vaccines, Synthetic / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001500:BCG Vaccine; D001426:Bacterial Proteins; C093276:ESAT-6 protein, Mycobacterium tuberculosis; D014614:Vaccines, Synthetic; D018664:Interleukin-12; D015847:Interleukin-4; D007371:Interferon-gamma; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D000217:Acyltransferases; C112580:antigen 85A, Mycobacterium tuberculosis
keywords: Immunogenicity; Protective efficacy; Recombinant bacille calmette-guerin; Tuberculosis