Specific cellular immunity and antitumor responses in C57BL/6 mice induced by DNA vaccine encoding murine AFP

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 th...

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Bibliographic Details
Published in:Hepatobiliary & pancreatic diseases international 2004-08, Vol.3 (3), p.440-443
Main Authors: Tian, Geng, Yi, Ji-Lin, Xiong, Ping
Format: Article
Language:English
Subjects:
alpha-Fetoproteins - genetics
alpha-Fetoproteins - immunology
Animals
Blotting, Western
Cancer Vaccines - genetics
Cancer Vaccines - immunology
Carcinoma, Hepatocellular - immunology
Carcinoma, Hepatocellular - therapy
DNA
Female
Immunity, Cellular - immunology
Immunoenzyme Techniques
immunotherapy
Kidney - physiology
Liver - physiology
Liver Neoplasms - immunology
Liver Neoplasms - therapy
Mice
Mice, Inbred C57BL
mouse
Plasmids
vaccine
Vaccines, DNA - genetics
Vaccines, DNA - immunology
α-fetoprotein
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Summary: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. 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. 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
ISSN:1499-3872