ABSTRACT
<p><b>OBJECTIVE</b>To construct the Escherichia coli (E. coli) prokaryotic expression system pET9aHPV11L2E7, purify the fusion protein L2E7 and study the immunnogenicity of the protein.</p><p><b>METHODS</b>The HPV11 L2, E7 coding region was amplified from condyloma acuminata tissue specimen by PCR. The recombinant plazmid pET9aHPV11L2E7 was established and sequenced. Fusion protein L2E7 (553 amino acids) was expressed in host strain BL21 (DE3plus) by IPTG inducing and identified by using SDS-PAGE and Western blotting. Then L2E7 protein purified with CM column was inoculated to Balb/c mice and its cell-mediated and humoral immunnogenicity was assessed by IFN-gamma enzyme-linked immunospot (ELISPOT) and enzyme-linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>The E. coli prokaryotic expression system pET9aHPV11L2E7 was established and the purified fusion protein L2E7 was obtained successfully. The mice in vivo experiment indicated that the purified protein L2E7 could induce HPV11E7 specific cell-mediated immune responses and high level HPV L2E7 antibody was detected in serum.</p><p><b>CONCLUSION</b>The purified fusion protein L2E7 could induce specific cell-mediated and humoral immune responses. It can be used as a candidate of genital wart immune therapeutic vaccine.</p>
Subject(s)
Animals , Female , Humans , Mice , Antibodies, Viral , Blood , Capsid Proteins , Genetics , Allergy and Immunology , Escherichia coli , Genetics , Metabolism , Gene Expression , Human papillomavirus 11 , Genetics , Allergy and Immunology , Mice, Inbred BALB C , Oncogene Proteins, Viral , Genetics , Allergy and Immunology , Papillomavirus Infections , Allergy and Immunology , Virology , Papillomavirus Vaccines , Genetics , Allergy and Immunology , Recombinant Fusion Proteins , Genetics , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To generate a human papillomavirus (HPV16) prophylactic and therapeutic vaccine candidate for cervical cancer.</p><p><b>METHODS</b>HPV16 major capsid protein L1 gene/minor capsid protein L2 gene and HPV16 early E6/E7 genes were inserted into a vaccinia virus expression vector. A strain of non-recombinant vaccinia virus containing the sequences was obtained through a homologous recombination and identified.</p><p><b>RESULTS</b>DNA hybridization confirmed that the HPV16L1/L2/E6/E7 genes were integrated into vaccinia virus DNA. Western Blot result showed that full-length L1/L2/E6/E7 proteins were co-expressed in CEF cells infected with the recombinant virus.</p><p><b>CONCLUSION</b>NTVJE6E7CKL1L2 could be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precancerous transformations.</p>