口服人乳头瘤病毒18 E7与次级淋巴组织趋化因子联合基因疫苗的抗肿瘤作用  

Anti-tumor effect of oral human papillomavirus 18 E7/secondary lymphoid chemokine combined gene vaccine

作　　者：曾凡慧[1,2] 张昌菊[2] 朱平[2] 雷小敏[3,4] 柳长柏[1] 

机构地区：[1]三峡大学分子生物学研究所,湖北宜昌443000 [2]三峡大学医学院,湖北宜昌443000 [3]三峡大学第二临床医学院 [4]三峡大学仁和医院,湖北宜昌443000

出　　处：《中国新药与临床杂志》2010年第1期40-44,共5页Chinese Journal of New Drugs and Clinical Remedies

基　　金：湖北省教育厅指导项目(13200613001)

摘　　要：目的观察口服人乳头瘤病毒(HPV)18型E7联合次级淋巴组织趋化因子(SLC)基因疫苗的抗肿瘤作用。方法Western blotting方法鉴定稳定表达E7的B16F1细胞株。小鼠腋下接种稳定表达E7的小鼠黑色素瘤B16F1细胞,荷瘤小鼠随机分为5组(n=12),灌胃途径给予各种伤寒沙门菌液免疫小鼠:生理盐水组(生理盐水0.2mL)、空载体组(携带空载体的菌液0.2mL)、E7组(携带E7的菌液0.2mL)、SLC组(携带SLC的菌液0.2mL)和E7联合SLC组(携带E7或SLC的菌液各0.1mL)。检测各组小鼠免疫后的瘤重及生存率、血清及阴道分泌物E7特异性抗体产生情况、脾细胞增殖指数和杀伤活性,并观察肿瘤组织局部淋巴细胞浸润情况。结果稳定表达E7的B16F1细胞株成功构建。E7联合SLC组小鼠瘤重显著降低,生存期显著延长(P<0.05),诱导产生血清E7特异性IgG抗体水平显著增加(P<0.01),淋巴细胞增殖指数和细胞毒T淋巴细胞杀伤率显著高于其他组(P<0.05或P<0.01)。SLC组及E7联合SLC组肿瘤组织浸润的淋巴细胞数均明显增多。结论口服HPV18型E7联合SLC基因疫苗可发挥抗肿瘤作用,其机制可能为增强体液和细胞免疫应答。AIM To observe the anti-tumor effect of oral human papillomavirus （HPV） 18 E7 /secondary lymphoid chemokine （SLC） combined gene vaccine.METHODS The successful construction of B16F1 cell lines expressed E7 was confirmed by Western Blotting.BALB /C mice were injected with B16F1 cells which expressed E7 in oxter,to establish mice tumor models.Then the mice were randomly divided into 5 groups （n=12 for each group）：normal saline group （normal saline 0.2 mL,ip）,empty vector group （Salmonella typhimurium SL3261 with empty vector 0.2 mL,ip）,E7 group （Salmonella typhimurium SL3261 with E7 0.2 mL,ip）,SLC group （Salmonella typhimurium SL3261 with SLC 0.2 mL,ip）,and E7 combined with SLC group （Salmonella typhimurium SL3261 with E7 0.1 mL ＋ Salmonella typhimurium SL3261 with SLC 0.1 mL,ip）.The tumor weight and survival time were recorded,the level of E7-specific antibody in serum and vaginal secretion were measured,the proliferation index of splenocytes and killing activities were calculated,and leukocytes infiltration were observed.RESULTS B16F1 cell line expressed E7 was successfully constructed.In the E7 combined with SLC group,the tumor weight decreased and survival time raised significantly （P 0.05）,the level of E7-specific IgG increased remarkably （P 0.01）,and the proliferation index of splenocytes and killing activity were notably higher than the other groups （P 0.05 or P 0.01）.High numbers of infiltrated leukocytes were observed in the E7 group as much as in the E7 combined with SLC group.CONCLUSION Oral HPV 18 E7 /SLC combined gene vaccine has the anti-tumor effect,which may be responsible for the enhancement of both humoral and cell-mediated immune response.

关 键 词：人乳头瘤病毒18 趋化因子 CC 疫苗 DNA 

分 类 号：R373.9[医药卫生—病原生物学]