Epitope analysis of cDNA sequences of Deinagkistrodon acutus snake venom metalloproteinases and observation of their immune protective effects / 第二军医大学学报

ABSTRACT

Objective: To analyze the epitopes of cDNA sequences of Deinagkistrodon acutus snake venom metalloproteinases using bioinformatical method, and to observe the immune protective effect of the new immunogen designed according to the identified epitopes. Methods: The cDNA sequences of Deinagkistrodon acutus snake venom metalloproteinases were amplified by PCR. The epitopes of the sequences were analyzed by Jameson-Wolf method and Clustal X software, then the sequences of the screened epitopes were artificially synthesized and linked to the vector pIRESneo. BALB/c mice were immunized by the resultant plasmid at 0, 2, and 4 weeks for three times, then the titers of the anti-serum were measured by ELISA. The immune protective effects of the anti-serum were tested by the neutralization of venom hemorrhagic activity and venom attacking test. Results: Bioinformatical analysis yielded 6 epitopes ( MPA-1-MPA-6). The ELISA results of anti-serum showed that these epitopes could induce immune reaction in mice, and the anti-serum was detectable even when it was diluted to 1100. The neutralization test and venom attacking test demonstrated that the anti-serum induced by the epitodes could neutralize the venom and protect the mice from haemorrhage. Conclusion: Six epitopes of Deinagkistrodon acutus snake venom metalloproteinases have been obtained successfully using bioinformatical method, and the new immunogen designed based on these epitopes shows a primary immune protective effect.