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Use of Interferon-γ Enzyme-linked Immunospot Assay to Characterize Novel T-cell Epitopes of Human Papillomavirus
Published on: March 8, 2012
[Construction of recombinant vaccina virus of HPV16 and its antitumor effect]
Wei Huang1, Hou-Wen Tian, Jiao Ren
1Department of Gynecology Oncology, The Affiliated Tumor Hospital, Guangxi Medical University, Nannning, Guangxi, 530021, PR China. huangwei@csco.org.cn
Background & Objective:
Cervical cancer is tightly related with high-risk types of human papillomavirus (HPV), among which HPV16 is most common. This study was to construct HPV16 L1, L2, E67 coexpression non-replication vaccina virus, and explore its immunization effects.
Methods:
HPV16 major capsid protein L1/L2 genes and HPV16 early E6/E7 genes were inserted into a vaccina virus expression vector to construct recombinant vaccina virus NTVJE67CKL1L2 by homologous recombination; the recombinant virus was identified by DNA hybridization and Western blot. C57BL/6 mice were immunized by NTVJE67CKL1L2; specific antibodies and specific cytotoxic T lymphocytes (CTLs) were detected. Immune protective effects of NTVJE67CKL1L2 on the mice were evaluated by challenges of TC-1 tumor cells.
Results:
HPV16 L1/L2/E6/E7 genes were integrated into vaccina virus DNA; Western blot confirmed that full-length L1/L2/E67 proteins were co-expressed in chicken embryo fibroblast (CEF) cells infected with the recombinant virus. NTVJE67CKL1L2 elicited specific antibodies against HPV16 L1/L2/E6/E7 and generated TC-1 cell-specific CTLs in mice. Mice boosted with NTVJE67CKL1L2 could tolerate the challenge of 1x10(4) TC-1 cells.
Conclusion:
NTVJE67CKL1L2 can be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precursor lesions.
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