[Enhancement of mouse immune response by recombinant adenovirus co-expressing VP1-2A of foot-and-mouth disease virus and porcine IFN-α].
OBJECTIVE
To construct the recombinant adenovirus co-expressing foot-and-mouth disease virus (FMDV) viral structural protein1-2A (VP1-2A) and porcine interferon-α (poIFN-α), and study its impact on mouse adaptive immune responses.


METHODS
FMDV VP1-2A genes and porcine IFN-α (poIFN-α) genes were successively cloned into pAdeno Vator-CMV5-IRES-GFP, and then both pAdeno Vator-CMV5-IRES-GFP with VP1-2A-poIFN-α and pAdeno Vator ΔE1/E3 were co-transfected into E.coli BJ5183 competent cells. The homologous recombinant plasmid rAd5VP1-2A-poIFN-α was screened and linearized by Pac I to expose the encapsidation signal and then transfected into HEK293A cells by Lipofectamine(TM)2000. The positive recombinant viruses were named rAd5VP1-2A-poIFN-α. The BALB/c mice were immunized by the recombinant adenovirus rAd5VP1-2A-poIFN-α or rAd5VP1 (adenoviral vectors solely expressing FMDV VP1) or blank adenovirus as control (Ad virus). The FMDV VP1-specific IgG, IgG1, IgG2a and cytokines IL-4, IFN-γ were determined by the ELISA, and the index of peripheral blood lymphocyte proliferation was detected by MTT assay.


RESULTS
The recombinant adenovirus co-expressing FMDV VP1-2A and porcine IFN-α (rAd5VP1-2A-poIFN-α) was successfully constructed. ELISA and MTT assay showed that compared with rAd5VP1 group, rAd5VP1-2A-poIFN-α could enhance the secretion of FMDV VP1-specific IgG, IgG1 and IgG2a, promote the secretion of IL-4 and IFN-γ, and increase the index of peripheral blood lymphocyte proliferation.


CONCLUSIONS
The rAd5VP1-2A-poIFN-α could significantly improve the mouse adaptive immune responses.
Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, School of Medical Examination, Ningxia Medical University, Yinchuan 750004, China.
Chunxia
C
Su
Department of Clinical Immunology and Laboratory, School of Medical Examination, Ningxia Medical University, Yinchuan 750004, China.
Xiangguo
X
Duan
Department of Agriculture Key Laboratory of Animal Diseases Diagnosis & Immunology, Nanjing Agricultural University, Nanjing 210095, China.
Ruibing
R
Cao
Department of Pathogen Biology and Immunology, School of Basic Medical Sciences, School of Medical Examination, Ningxia Medical University, Yinchuan 750004, China.
Jie
J
Zheng
Department of Agriculture Key Laboratory of Animal Diseases Diagnosis & Immunology, Nanjing Agricultural University, Nanjing 210095, China.
Puyan
P
Chen
D056704:Adaptive Immunity; D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D036022:Capsid Proteins / Q000235:genetics* / Q000276:immunology; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D016898:Interferon-alpha / Q000235:genetics*; D007371:Interferon-gamma / Q000097:blood; D015847:Interleukin-4 / Q000097:blood; D051379:Mice; D008807:Mice, Inbred BALB C; D013552:Swine; D014614:Vaccines, Synthetic / Q000276:immunology; D014765:Viral Vaccines / Q000276:immunology*
D000914:Antibodies, Viral; D036022:Capsid Proteins; D016898:Interferon-alpha; C442501:VP1 protein, Foot-and-mouth disease virus; D014614:Vaccines, Synthetic; D014765:Viral Vaccines; D015847:Interleukin-4; D007371:Interferon-gamma