[Construction of human metapneumovirus DNA vaccine and study on its immune response in mice].
OBJECTIVE
To construct human metapneumovirus (hMPV) DNA vaccines and evaluate the cellular and humoral immune response in mice.


METHODS
Fusion protein FdeltaTM (without transmembrane domain) gene and M gene of hMPV were amplified from cDNA by PCR, then DNA vaccines pcDNA3.1His-FdeltaTM and pcDNA3.1His-M were constructed to verify the expression of F and M protein by Western blotting and indirect immunofluorescent assay (IFA) respectively. Serum IgG and spleen cell CTL were detected with ELISA and ELISPOT assay after the BALB/c mice were immunized intramuscularly with the vaccines.


RESULTS
The candidate DNA vaccines could express FdeltaTM and M protein as detected with Western blotting and IFA. The IgG antibody titers of mice was 1:44 when immunized with pcDNA3.1His-FdeltaTM, but could increase to 1:64 when co-immunized with pcDNA3.1His-M. ELISPOT assay demonstrated that IFN-gamma-secreting effector T cells reached 42 +/- 8.9 in co-immunization group, higher than single vaccine pcDNA3.1His-FdeltaTM group (32 +/- 7.4).


CONCLUSIONS
DNA vaccine pcDNA3.1His-FdeltaTM could induce specific cellular and humoral immune responses, and the immune response could increase when co-immunization with pcDNA3.1His-M was carried out.
Liu
Wen-pei
WP
State Key Laboratory for Molecular Virology and Genetic Engineering, National Institute for Viral Disease Control and Prevention, China CDC, Beijing 100052, China.
Zheng
Li-shu
LS
Duan
Zhao-jun
ZJ
Xie
Zhi-ping
ZP
Zhang
Qian
Q
Zhang
Wan-ju
WJ
Hou
Yun-de
YD
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D005260:Female; D006801:Humans; D007114:Immunization; D029121:Metapneumovirus / Q000235:genetics / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D018184:Paramyxoviridae Infections / Q000276:immunology* / Q000517:prevention & control / Q000821:virology; D019444:Vaccines, DNA / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology*; D014764:Viral Proteins / Q000008:administration & dosage / Q000235:genetics / Q000276:immunology; D014765:Viral Vaccines / Q000008:administration & dosage / Q000235:genetics* / Q000276:immunology*
D000914:Antibodies, Viral; D019444:Vaccines, DNA; D014764:Viral Proteins; D014765:Viral Vaccines