Characterization of Antibody Generated Against GFP Protein Subsequent to cDNA Injection into Rat Femoral Muscle

概要

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In vivo injection of complementary DNA (cDNA) encoding for a specific protein has recently been shown to induce immune response to the antigenic protein. The kinetics of antibody inducement with cDNA have yet to be fully determined. The present study was conducted so as to characterize this induction and comparison was made with conventional antibody formation using protein as antigen. Rats were injected with 50μg pEGFP-N1 plasmid DNA in 50μ1 PBS, eight times at one-week interval in the right femoral muscle or 50μg recombinant GFP protein in 100μ1 PBS, 2 times at 4-week interval intraperitoneally. Blood was collected from the tail artery 6 times at two-week interval up to 10 weeks under anesthesia. The induced antibodies were specified using Westrn blotting and quantitated with enzyme linked immunosorbent assay (ELISA). In the recombinant protein-injected rat groups, anti-GFP lgG 1, but no lgG 2b, was detected in the serum, while in the plasmid DNA-injected group, lgG 2b, but not lgG 1, was noted to be the major product induced. The extent of lgG 2a inducement was essentially the same for the cDNA and protein injected groups. Intramuscular DNA injection would thus appear quite likely to induce antibody formation by modulating Th1-type helper T cells in contrast to Th2 cells when antibody is produced through the action of protein as antigen.
北里大学の論文
2002-06-30