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.
出处
《南昌大学学报（医学版）》
CAS
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)
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
human papillomavirus type 16
human beta-defensin-2
ET-specific antibodies
serum
cellular immunity
animals,laboratory
mice
