[Immunity and EGFPC3 expression of recombinant attenuated salmonella typhimurium carrying plasmid pcDNA3s and EGFPC3 in mice].
OBJECTIVE
To study the immune characterization of HBsAg DNA recombinant attenuated salmonella typhimurium vaccine and the expression of enhanced green fluorescence protein gene pEGFPC3 in mice.


METHODS
Plasmid pcDNA3s and pEGFPC3 was transformed into attenuated salmonella typhimurium SL8786 by electricity instrument to construct recombinant vaccine SL8786/EGFPC3 and SL8786/pcDNA3s. Mice were immunized with the recombinant vaccine and the quantity of anti-HBs antibody in the sera was measured by time resolved fluorescence immunoassay(TRFIA), While the cytotoxocity of CTL was measured by LDH release assay.


RESULTS
It produced strong specificity CTL response after taking SL8786/pcDNA3s orally to mice and anti-HBs was highest in 11 weeks. EGFPC3 expression in two mice spleen cells was detected by flow cytometry(FCM). EGFP masculine ratio is 19.20% and 17.36% respectively. but SL8786/pcDNA3 in two mice spleens cell EGFP masculine ratio only is 1.95% and 1.63%. The mice was taken SL8786/EGFPC3 orally after three weeks, fluorescence expression in its liver, spleen, kidney, chest gland, duodenum, muscle, etc. were observed.


CONCLUSIONS
HBV DNA vaccine can induce specific cellular and humoral immune responses in vivo in mice by oral. Recombinant attenuated salmonella typhimurium SL8786/EGFPC3 we constructed. It has green fluorescence expression in mice's partial organization, and it offers a model for research gene engineering vaccine.
Fang
Xi-Xiu
XX
Food Nutrition and Safety Institute, Food Science and Technology College, Key Laboratory of Industry's Biotechnology of Education's Ministry, Southern Yangtze University 214036, China.
Wang
Dong-Mei
DM
Shi
Yong-Hui
YH
LE
Guo-Wei
GW
Bai
Xin-Peng
XP
Liu
Jian-Wen
JW
D000818:Animals; D000917:Antibody Formation / Q000235:genetics / Q000276:immunology; D018274:Electroporation; D005260:Female; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D049452:Green Fluorescent Proteins / Q000235:genetics* / Q000378:metabolism; D006514:Hepatitis B Surface Antigens / Q000235:genetics; D007111:Immunity, Cellular / Q000235:genetics / Q000276:immunology; D008297:Male; D051379:Mice; D010957:Plasmids / Q000235:genetics; D012486:Salmonella typhimurium / Q000235:genetics*; D013154:Spleen / Q000378:metabolism; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology; D014162:Transfection; D014613:Vaccines, Attenuated / Q000235:genetics / Q000276:immunology; D019444:Vaccines, DNA / Q000235:genetics* / Q000276:immunology*; D014614:Vaccines, Synthetic / Q000235:genetics / Q000276:immunology
D006514:Hepatitis B Surface Antigens; D014613:Vaccines, Attenuated; D019444:Vaccines, DNA; D014614:Vaccines, Synthetic; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins