治疗性HPV16ZHsp65E6E7无佐剂重组蛋白疫苗的实验研究.doc

治疗性HPV16Z-Hsp65-E6/E7无佐剂重组蛋白疫苗的实验研究 [摘要] 目的：研究治疗性HPV16 Z-Hsp65-E6/E7重组蛋白疫苗的抗肿瘤相关生物活性。方法：通过淋巴细胞增殖实验和细胞毒性杀伤实验研究该疫苗激发的细胞免疫反应及反应强度；研究该疫苗对小鼠TC-1肿瘤细胞移植瘤的体内治疗作用和对小鼠生存期的影响。结果：经检测证实该重组蛋白疫苗免疫小鼠后小鼠脾淋巴细胞增殖明显，并可特异性地在体外杀伤TC-1细胞，能诱导特异性的细胞免疫反应；动物体内抑瘤试验研究显示该疫苗对HPV16病毒转化的TC-1细胞小鼠移植瘤有显著的治疗作用。结论：体外实验证实该疫苗有较强的免疫原性，能激发特异性细胞免疫反应；体内显著抑制HPV16转化的TC-1肿瘤细胞生长。治疗性HPV16 Z-Hsp65-E6/E7重组蛋白疫苗的初步研究证实该疫苗的进一步研究将对HPV16病毒感染相关疾病的治疗具有实际应用价值。[关键词] 人乳头瘤病毒16型；治疗性疫苗；宫颈癌 The experiment study of Therapeutic Adjuvant-free protein Vaccine—HPV16Z-Hsp65-E6/E7 [abstract] Objective: To study the biological effects of the HPV16Z-Hsp65-E6/E7 fusion protein vaccine on the tumor associated with HPV16 infection. Methods: We tested the cellular immune responsive intensity to the vaccine by the lymphocyte proliferation and CTL response and studied the therapeutic effect of the vaccine on mouse TC-1 cell transplanted cancer in vivo, and the influence on mouse lifetime . Results: The spleen lymphocytes from the C57BL/6 mouse immunized by the Hsp65-E6/E7 vaccine could be proliferated obviously in the presence of the protein and TC-1 tumor cell could be lysed specifically by immune activated lymphocytes In vitro. The animal therapeutic experiment in vivo showed that the vaccine could suppress the growth of TC-1 cell transplanted tumor remarkably. Conclusions: The recombined vaccine could induce specific cellular immune response in vitro and suppress HPV16 positive TC-1 tumor cell growth obviously in vivo. The preliminary study of the adjuvant-free vaccine confirmed that the further research and development of this vaccine may have the practical application value to treating the HPV16 correlative disease. [key words]: Human Papillomavirus 16 Therapeutic vaccine Cervical cancer ??? HPV与多种人类上皮来源的恶性肿瘤和不典型增生的发生和发展关系密切，高危HPV16型的感染是女性宫颈癌形成的主要原因[1]。在我国妇女宫颈癌组织中HPV感染检出率大于99%，其中HPV16占60%以上[2]。世界范围内每年有40多万新发宫颈癌病例，约20万人死于该病，位居女性肿瘤死亡人数的第二位[3]。目前宫颈癌的治疗主要是手术和放疗，其治疗5年生存率仅50%左右，另外对宫颈的早期病变和HPV感染相关的疾病的治疗尚缺乏理想的方法。因此研制新的药物用于治疗HPV感染尤其是HPV16型引起的相关疾