ABSTRACT
<p><b>OBJECTIVE</b>To identify specific T lymphocyte epitope on E6 protein of human papillomavirus type 18 in mice.</p><p><b>METHODS</b>Infection with one recombinant vaccinia virus rVVJ18 E7, E6 respectively in C57 BL/6 and BALB/c mice, specific cellular immune responses were detected by ELISPOT or intracellular cytokine stainings by using a series of overlapping synthetic peptides covering full length of the amino acid sequence of E6 and E7 proteins or various truncated peptides.</p><p><b>RESULTS</b>The rVVJ18 E7, E6 generated significant E6 specific T-cell immune responses in vaccinated mice. Mapping of the epitope of E6 revealed that the peptides E6(67-75 ( KCIDFYSRI) and E6(60-68) (IPHAAGHKC) presented respectively by C57BL/6 and BALB/c mice were the optimal peptides to activate E6-specific CD8+ T lymphocytes. However no positive cellular immune responses stimulated with various E7 peptides were detected by ELISPOT in immunized mice.</p><p><b>CONCLUSIONS</b>Two specific T lymphocyte epitopes were identified on E6 protein in C57BL/6 and BALB/c mice, which will provide the basis to evaluate cellular immune response elicited by HPV18 E6 protein based vaccine.</p>
Subject(s)
Animals , Humans , Mice , CD8-Positive T-Lymphocytes , Allergy and Immunology , DNA-Binding Proteins , Allergy and Immunology , Epitopes, T-Lymphocyte , Allergy and Immunology , Human papillomavirus 18 , Allergy and Immunology , Mice, Inbred BALB C , Mice, Inbred C57BL , Oncogene Proteins, Viral , Allergy and Immunology , Papillomavirus Vaccines , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To express HPV6bL2deltaN360E7E6 fusion protein in E. coli and preliminarily evaluate its immune effect.</p><p><b>METHODS</b>Three HPV6b gene fragments, which were L2(1-360 bp), E7 and E6, were fused by overlapping PCR, then were inserted into a prokaryotic expression vector and expressed in E. coli. C57BL/6 mice were immunized with purified fusion protein plus Al (OH)3 and/or CpG adjuvants through intramuscular route, the cellular and humoral immune responses were detected by IFN-gamma ELISPOT and ELISA respectively.</p><p><b>RESULTS</b>Protein plus CpG adjuvant could induce the strongest cellular immune response to E7 and E6, high antibody titer against L2 could be detected in all immunized groups but there were no significant difference among these groups.</p><p><b>CONCLUSIONS</b>HPV6bL2deltaN360E7E6 gene was successfully cloned into pQE30 vector and expressed in E. coli, the fusion protein was also purified and proved that could induce strong cellular and humoral immune responses with appropriate adjuvant in C57BL/ 6 mice and could be used for future research.</p>