Specific cellular immunity and antitumor responses in C57BL/6 mice induced by DNA vaccine encoding murine AFP.
BACKGROUND
The inoculation of plasmid DNA encoding tumor-associated antigens is a novel and powerful strategy for antitumor vaccination. This study was designed to construct the DNA vaccine of mouse AFP and to observe the specific cellular immunologic responses and the antitumor responses in mice induced by this vaccine.


METHODS
The murine AFP gene was amplified by RT-PCR from total RNA extracted from Hepa1-6 cells and cloned into the vector pcDNA3.1 to construct pmAFP. The DNA vaccine was identified by restriction enzyme analysis, sequencing and expression. EL-4 (mAFP) was developed by stable transfection of EL-4 cells with pmAFP. The frequency of cells producing IFN-gamma in splenocytes harvested from the mice immunized with the DNA vaccine by intramuscular injection was measured by enzyme linked immunospot (ELISPOT). The mice immunized with the DNA vaccine were inoculated with EL-4 (mAFP) cells in back to observe the inhibitory effect of the immunization on tumor. On the other hand, blood samples were collected from the immunized mice to check the functions of the liver and kidney.


RESULTS
The murine AFP gene was successfully cloned by RT-PCR. Results from restriction enzyme analysis, sequencing and expression showed that the DNA vaccine was successfully constructed. The expression of mAFP mRNA in EL-4 (mAFP) was confirmed by RT-PCR. The results of ELISPOT showed that the number of IFN-gamma- producing cells of the pmAFP vaccine group was significantly higher than that of other groups (P<0.01). The tumor volume in the pmAFP vaccine group (1042.42+/-123.71 mm(3)) was significantly smaller than that in other groups (P<0.01). The function of mouse liver and kidney in each group was unchanged.


CONCLUSIONS
The successfully constructed DNA vaccine of AFP can induce specific cellular immunologic responses and significant antitumor responses in mouse and has no impact on the function of mouse liver and kidney.
Tian
Geng
G
Department of Oncology, Shenzhen Second Hospital, Shenzhen 518035, China. geng_tian707@hotmail.com
Yi
Ji-Lin
JL
Xiong
Ping
P
D000818:Animals; D015153:Blotting, Western; D019496:Cancer Vaccines / Q000235:genetics / Q000276:immunology*; D006528:Carcinoma, Hepatocellular / Q000276:immunology / Q000628:therapy*; D005260:Female; D007111:Immunity, Cellular / Q000276:immunology*; D007124:Immunoenzyme Techniques; D007668:Kidney / Q000502:physiology; D008099:Liver / Q000502:physiology; D008113:Liver Neoplasms / Q000276:immunology / Q000628:therapy*; D051379:Mice; D008810:Mice, Inbred C57BL; D010957:Plasmids; D019444:Vaccines, DNA / Q000235:genetics / Q000276:immunology*; D000509:alpha-Fetoproteins / Q000235:genetics / Q000276:immunology*
D019496:Cancer Vaccines; D019444:Vaccines, DNA; D000509:alpha-Fetoproteins