CAJ | 학술논문

目的利用重组HPV16 L1 DNA真核表达质粒免疫C57BL/6小鼠探讨DNA疫苗的免疫效果.方法实验动物C57BL/6随机分为实验组(pcDNAL1)和对照组(pcDNA3.1和PBS),用免疫原共免疫三次,间隔三周,末次免疫后10-14天行足垫肿胀试验检测DTH反应;之后,拉颈处死,尸检观察.行脾细胞抗原特异性增殖反应和CD4/CD8,IFN-γ+或IL10+细胞FACS分析.内脏用于组织学检查.结果质粒pcDNAL1组脾细胞增殖指数、足垫肿胀程度、CD8+IFN-γ+细胞均高于对照组.整个免疫过程中动物无行为异常,尸检未见异常病变.结论裸pcDNAL1 质粒免疫C57BL/6小鼠能诱发特异性细胞免疫反应,且安全性较高.这将为其进一步临床试验奠定牢固的理论和实践基础.
목적이용중조HPV16 L1 DNA진핵표체질립면역C57BL/6소서탐토DNA역묘적면역효과.방법실험동물C57BL/6수궤분위실험조(pcDNAL1)화대조조(pcDNA3.1화PBS),용면역원공면역삼차,간격삼주,말차면역후10-14천행족점종창시험검측DTH반응;지후,랍경처사,시검관찰.행비세포항원특이성증식반응화CD4/CD8,IFN-γ+혹IL10+세포FACS분석.내장용우조직학검사.결과질립pcDNAL1조비세포증식지수、족점종창정도、CD8+IFN-γ+세포균고우대조조.정개면역과정중동물무행위이상,시검미견이상병변.결론라pcDNAL1 질립면역C57BL/6소서능유발특이성세포면역반응,차안전성교고.저장위기진일보림상시험전정뢰고적이론화실천기출.
Objective To investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine. Methods C57BL/6 mice were randomly divided into 3 groups: experimental groupⅠ (treated with pcDNA L1), control group Ⅱ (treated with pcDNA3.1) and control group Ⅲ (treated with PBS buffer). The mice were immunized three times during a three-week interval. Ten to fourteen days after the third inoculation, a footpad swelling test was used to detect delayed-type hypersensitivity (DTH) responses. Antigen-specific splenocyte proliferation assay and quantitation of IFN-γ cells in splenocytes were performed by FACS assay. Results In the experimental group, splenocytes actively proliferated after stimulation with HPV16 VLP, and had developed a markedly larger amount of CD8+ IFN-γ+ cells, which is an index for special CTL. Also, the footpad was significantly thickened upon inoculation with HPV16 VLP. Conclusion Naked DNA vaccine of HPV16 L1 can induce specific cell-mediated immune responses in mice, which should be considered for evaluation of HPV16 DNA vaccine feasibility.