摘要
背景与目的大量研究表明,宫颈癌的发生与高危型人乳头状瘤病毒(humanpapillomavirus,HPV)的感染密切相关,其中以HPV16为主。本研究以非复制型重组痘苗病毒为载体,构建共表达HPV16L1、L2、E67蛋白的重组非复制型痘苗病毒,并检测其免疫效果。方法以痘苗病毒为载体,利用同源重组技术构建共表达HPV16L1、L2、E67基因的重组痘苗病毒NTVJE67CKL1L2。用该病毒免疫C57BL/6小鼠后,检测其特异性抗体和特异性的细胞毒性T淋巴细胞(cytotoxicTlymphocyte,CTL);以TC-1肿瘤细胞攻击经免疫后的小鼠,观察其免疫保护效果。结果经斑点杂交结果显示重组病毒基因组中有L1、L2、E6、E7基因插入;经Westernblot检测,重组病毒能稳定表达HPV16L1、L2、E67蛋白。动物实验结果表明,NTVJE67CKL1L2在C57BL/6小鼠体内可诱发L1、L2、E67特异性抗体产生;可诱发产生针对TC-1细胞的特异性的CTL反应;加强免疫后的小鼠能耐受1×104个TC-1肿瘤细胞的攻击。结论非复制型重组痘苗病毒NTVJE67CKL1L2可以作为对HPV16相关肿瘤及其癌前病变进行免疫治疗的候选疫苗。
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 1×104 TC-1 cells. ONCLUSION: NTVJE67CKL1L2 can be taken as a candidate of prophylactic and therapeutic vaccine for HPV-associated tumors and their precursor lesions.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2005年第7期817-822,共6页
Chinese Journal of Cancer
