人β防御素-2联合HPV16 E7c免疫小鼠的免疫学效应实验研究  

Immunological Effects of Human Beta-defensin 2 Combined with HPV16 E7c Fragment in BALB/c Mice

作　　者：罗丹青[1] 苏虹[2] 曾文兴[2] 邓玲[3] 张志勇[4] 王美珍[5] 李东旭[1] 王福财[2] 刘玉琳[2] 施桥发[2] 

机构地区：[1]南昌大学医学院,2009级南昌330006 [2]南昌大学基础医学院免疫学教研室,南昌330006 [3]南昌大学抚州分院护理系,江西抚州344000 [4]江西省人民医院检验科,南昌330006 [5]基础医学院免疫学教研室,南昌330006

出　　处：《南昌大学学报（医学版）》2013年第1期8-12,共5页Journal of Nanchang University：Medical Sciences

基　　金：国家自然科学基金(30760280);国家大学生创新性实验计划项目(101040304);江西省科技厅社会发展重点项目(2007BS12802);江西省卫生厅科技计划项目(20072016);江西省教育厅科技计划项目(GJJ08126)

摘　　要：目的探讨以人β防御素-2为佐剂的HPV16E7c免疫功能及细胞免疫应答。方法将30只小鼠按随机数字表法分为3组:PBS组、pcDNA3.1(+)/E7c组、pEGFP-C1/HBD2-E7c组,每组10只。PBS组采用PBS100μg·只-1行股四头肌肌内注射,进行初次免疫;pcDNA3.1(+)/E7c组采用pcDNA3.1(+)/E7c质粒100μg·只-1行股四头肌肌内注射,进行初次免疫;pEGFP-C1/HBD2-E7c组采用pEGFP-C1/HBD2-E7c联合质粒100μg·只-1行股四头肌肌内注射,进行初次免疫。3组初次免疫后,再间隔2周,以上述方法和剂量加强免疫2次。共免疫3次。3组小鼠免疫1周后,断颈处死小鼠,取小鼠脾脏称重,计算脾脏脏器指数及计数脾淋巴细胞数量,流式细胞仪检测淋巴细胞亚群CD4+、CD8+细胞数量及比值;采用摘眼球采血法收集小鼠血清,采用ELISA法检测小鼠血清E7抗体水平。结果 pcDNA3.1(+)/E7c组、pEGFP-C1/HBD2-E7c组小鼠脾脏脏器指数均明显低于PBS组(均P<0.01)。pEGFP-C1/HBD2-E7c组、pcDNA3.1(+)/E7c组小鼠CD4+、CD8+阳性细胞比例、绝对数均明显高于PBS组(均P<0.05),pEGFP-C1/HBD2-E7c组、pcDNA3.1(+)/E7c组小鼠CD4+/CD8+比值均明显低于PBS组(均P<0.05),pEGFP-C1/HBD2-E7c组、pcDNA3.1(+)/E7c组小鼠血清E7抗体水平均明显高于PBS组(均P<0.01),pEGFP-C1/HBD2-E7c组小鼠血清E7抗体水平明显高于pcDNA3.1(+)/E7c组(P<0.05)。结论人β防御素-2可明显改善HPV16E7c基因的免疫原性,提高HPV16E7特异性抗体水平,升高免疫小鼠脾CD4+和CD8+T淋巴细胞数量。with human Objective To papillomavirus Methods Thirty BALB/c explore the immunological effects of human beta-defensin 2 combined type 16 E7c and the cellular immune responses in BALB/c mice. mice were randomly divided into three groups, with 10 mice in eachgroup. Mice in PBS group,pcDNA3.1（＋）/E7c group and pEGFP-C1/HBD2-E7c group were primarily immunized into quadriceps muscle by intramuscular injection of PBS 100 μg, pcDNA3. 1（＋）/E7c plasmid 100μg and pEGFP-C1/HBD2-E7c plasmid 100/ag, respectively. Booster immunization was performed two times 2 weeks after primary immunization. Mice were sacrificed bycervical dislocation at 7 days after the last immunization and spleens were weighed to calculate the spleen index. The number of spleen lymphocytes was measured, and number of subsets of CD4＋and CD8＋ cells and CD4＋/CD8＋ ratio were determined by flow cytometry. Serum samples were collected by eyeball removal and E7-specific antibodies were measured by ELISA. Results Cornpared with PBS group,the spleen index and CD4＋/CD8＋ ratio significantly decreased but the pro- portion and number of CD4＋ and CD8＋ cells and serum levels of ET-specific antibodies obviouslyincreased in both pEGFP-C1/HBD2-E7c group and pcDNA3.1 （＋）/E7c group（P〈0.05 or P〈0.01）. Compared with pcDNA3.1 （＋）/E7e group, serum levels of E7-speeific antibodies remark-ably increased in pEGFP-C1/HBD2-E7c group（P〈0. 05）. Conclusion Human-beta defensin 2 can improve the immunogenicity of HPV16 ETc gene,enhance the levels of E7-specific antibodiesand increase the number of CD4＋ and CD8＋ T lymphocytes.

关 键 词：人乳头瘤病毒16型 人Β防御素-2 E7抗体 血清 细胞免疫 动物 实验 小鼠 

分 类 号：R391[医药卫生—基础医学] R-332