[Transient expression of Echinococcus granulosus Eg95 DNA vaccine and induction of immune response in mice].
OBJECTIVE
To detect the in vitro expression of pcDNA3-Eg95 and to observe the immunological effect of the Eg95 DNA vaccine in mice.


METHODS
The eukaryotic recombinant plasmid pcDNA3-Eg95 was transfected into HeLa cells with liposome-mediated method. RT-PCR, ELISA and Western blotting were used to analyze the expression of Eg95 mRNA and Eg95 protein, respectively. The BALB/c mice were immunized by pcDNA3-Eg95. Anti-Eg95 IgG and IgG2a in murine serum were determined by ELISA. The proliferation activity of spleen T lymphocytes was tested using MTT assay.


RESULTS
Using RT-PCR method, the expression of Eg95 mRNA was confirmed in vitro. The results of ELISA and Western blotting showed that there was a specific Eg95 protein, which can be specifically recognized by anti-sera of Eg95-prokaryotic-expressing protein in pcDNA3-Eg95 transfected HeLa cell lysis. The specific IgG was induced during the 3rd week and continued to increase until week 10. IgG2a was detected after 2 weeks and maintained a higher level till week 10. There was a significant difference of IgG2a level between pcDNA3-Eg95 immunized group and pcDNA3 control (P<0.01). In spleen T cell proliferation response, the stimulation index (SI) in pcDNA3-Eg95 group was higher than that of vector control (P<0.01).


CONCLUSIONS
Eg95 DNA vaccine can induce significant cellular and humoral immune response in mice.
Lin
Ren-yong
RY
Xinjiang Hydatid Fundamental Medical Key Lab, First Teaching Hospital of Xinjiang Medical University, Urumqi 830054, China.
Ding
Jian-bing
JB
Lu
Xiao-mei
XM
Wang
Xiao-feng
XF
Arziguli
Wei
Xiao-li
XL
Wang
Yan
Y
Wen
Hao
H
D000818:Animals; D000909:Antibodies, Helminth / Q000097:blood; D000947:Antigens, Helminth / Q000096:biosynthesis / Q000235:genetics / Q000276:immunology*; D003001:Cloning, Molecular; D004443:Echinococcosis / Q000276:immunology / Q000517:prevention & control; D048209:Echinococcus granulosus / Q000276:immunology; D005260:Female; D006367:HeLa Cells; D015801:Helminth Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000276:immunology*; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D019444:Vaccines, DNA / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000276:immunology
D000909:Antibodies, Helminth; D000947:Antigens, Helminth; C446034:EG95 protein, Echinococcus granulosus; D015801:Helminth Proteins; D019444:Vaccines, DNA; D014614:Vaccines, Synthetic