[Construction of the eukaryotic expression vector with IL-2 gene and VP2 gene of PPV and research on immunogenicity].
To construct gene vaccine of PPV and to investigate the effects of interleukin 2 (IL-2) as an adjuvant on immune responses in mouse, the recombinant expression plasmid of pCIneo-IL2-VP2 was constructed and transfected into PK-15 cells by lipofectamine, the expressed product was detected by immunofluore assay. To study the immune effects of DNA vaccine in vitro and in vivo, mice were used as the animal model. The recombinant plasmid pCIneo-IL2-VP2, the control plasmid pCI-neo and the PPV live vaccine were immunized by intramuscular injection. Anti-PPV antibodies were measured by ELISA, lymphocyte proliferation activity was detected using MTT method, and the specific killing activities of CTL were assayed too. The results show that the immunized mice produced PPV antibody after one week, and reached to highest after four weeks. Compared with the control group, the pCIneo-IL2-VP2 immunized group produced significant differences in the antibody titers, the lymphocyte proliferation activity and the specific killing activities of CTL. The pCIneo-IL2-VP2 induced humoral and cellular immunity responses similarly to that the live vaccine induced. These results manifested that the PPV DNA vaccine successfully induced humoral and cellular immunity response in mice with the IL-2 gene as an adjuvant.
Cui
Bao-An
BA
Henan Key Laboratory for Animal Food Safety, Zhengzhou, Henan 450002, China. baoancui@henau.edu.cn
Wei
Zhan-Yong
ZY
Wang
Xue-Bin
XB
Huang
Ke-He
KH
Jin
Xi-Xin
XX
Dong
Zhen-Jie
ZJ
Zheng
Lan-Lan
LL
D000276:Adjuvants, Immunologic / Q000235:genetics; D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D000956:Antigens, Viral / Q000235:genetics / Q000276:immunology*; D036022:Capsid Proteins / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors; D007114:Immunization; D007376:Interleukin-2 / Q000235:genetics / Q000276:immunology*; D051379:Mice; D028323:Parvovirus, Porcine / Q000235:genetics / Q000276:immunology; D011897:Random Allocation; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000276:immunology*; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection; D019444:Vaccines, DNA / Q000276:immunology*; D014765:Viral Vaccines / Q000276:immunology*
D000276:Adjuvants, Immunologic; D000914:Antibodies, Viral; D000956:Antigens, Viral; D036022:Capsid Proteins; D007376:Interleukin-2; D011993:Recombinant Fusion Proteins; C528774:VP2 protein, porcine parvovirus; D019444:Vaccines, DNA; D014765:Viral Vaccines