HPV-16 E6E7抗原表位嵌合体DNA疫苗黏膜免疫抑制宫颈癌的效果研究

Study on the effect of mucosal immunosuppression of cervical cancer by HPV-16 E6E7 antigenic epitope chimeric DNA vaccine

目的 评价泛素及热休克蛋白70(HSP 70)C融合人乳头瘤病毒(HPV)16型E6E7表位嵌合体DNA疫苗鼻内黏膜免疫对宫颈癌移植瘤的预防和治疗效果。方法 建立TC-1小鼠肿瘤模型,以壳聚糖为发送载体,通过滴鼻免疫给药免疫C57BL/6小鼠。MTT法和淋巴毒性T细胞(CTL)反应检测小鼠脾脏T淋巴细胞增殖、流式细胞术检测细胞因子表达水平、肿瘤生长曲线和荷瘤小鼠存活时间评价壳聚糖包裹的HPV-16 E6E7嵌合体DNA疫苗鼻内黏膜免疫后对HPV-16型相关宫颈癌移植瘤的预防和治疗效果。结果 与对照组pcD-UH相比,实验组pcD-UE和pcD-UEH均具有显著的CTL反应,延缓HPV-16型相关宫颈癌移植瘤生长,但对已生成肿瘤无影响。鼻内黏膜途径发送壳聚糖包裹HPV-16 E6E7抗原表位嵌合体DNA疫苗仅能产生较弱的细胞免疫应答。结论 HPV-16 E6E7抗原表位嵌合体DNA疫苗通过鼻腔免疫,具有一定的肿瘤治疗和显著的肿瘤预防效果,为新型HPV疫苗的研制奠定了基础。

Objective To evaluate the preventive and therapeutic efficacy of intranasal mucosal immunization with ubiquitin and heat shock protein 70(HSP 70) C fusion human papillomavirus(HPV) type 16 E6E7 epitope chimeric DNA vaccine against cervical cancer graft tumors.Methods TC-1 mouse tumor model was established,and chitosan was used as the delivery vehicle to immunize C57BL/6 mice by nasal drop immunization administration.MTT assay and cytotoxic T lymphocyte(CTL) reaction were used to detect the proliferation of mouse spleen T-lymphocytes,cytokine expression levels were detected by flow cytometry,and the tumor growth curves and the survival time of the loaded mice were evaluated to determine the preventive and therapeutic effects of chitosan-encapsulated HPV-16 E6E7 chimeric DNA vaccine after intranasal preventive and therapeutic effects on HPV-16-associated cervical cancer graft tumors after mucosal immunization.Results Compared with control pcD-UH,both pcD-UE and pcD-UEH in the experimental group had a significant CTL response and delayed the growth of transplanted tumors in HPV-16-associated cervical cancer,but had no effect on generated tumors.The intranasal mucosal route sending chitosan-coated HPV-16 E6E7 antigenic epitope chimeric DNA vaccine produced only a weak cellular immune response.Conclusion The HPV-16 E6E7 antigen epitope chimeric DNA vaccine has certain tumor treatment and significant tumor prevention effects through nasal immunity,laying the foundation for the development of a new HPV vaccine.