单纯疱疹病毒2gD基因的树突细胞疫苗免疫效应研究  

An experimental study on the immune response induced by a dendritic cell-based vaccine carrying the herpes simplex virus type 2 glycoprotein D gene in mice

作　　者：樊建勇[1] 王颖[1] 杨慧兰[1] 梁洁[1] 李翠华[1] 

机构地区：[1]广州军区广州总医院皮肤科,510010

出　　处：《中华皮肤科杂志》2014年第8期574-577,共4页Chinese Journal of Dermatology

基　　金：国家自然科学基金（81071401）

摘　　要：目的 观察单纯疱疹病毒2 gD（HSV-2 gD）基因的树突细胞（DC）疫苗在动物实验中诱发特异性免疫应答情况.方法 用腺病毒介导的、HSV-2 gD基因修饰的DC（pAdeno-HSV-2 gD-DC）疫苗免疫BALB/c小鼠3次,末次免疫后第10天检测小鼠血清中gD IgG水平;同时,分离小鼠脾淋巴细胞,用HSV-2gD蛋白刺激,MTT法检测小鼠脾淋巴细胞增殖情况、乳酸脱氢酶释放法测定细胞毒性T淋巴细胞（CTL）活性及ELISA法检测脾淋巴细胞培养上清中干扰素γ（IFN-γ）、白介素4（IL-4）水平.实验分为4组,pAdeno-DC组、pAdeno-HSV-2 gD-DC组、DC组、空白对照组,每组10只.结果 pAdeno-HSV-2 gD-DC组小鼠血清内可检测到高滴度针对HSV-2gD蛋白的抗体（A450值为0.313±0.034）,与pAdeno-DC组（0.034±0.009）、DC组（0.028±0.009）、空白对照组（0.026±0.010）比较,差异有统计学意义（P＜0.05）.pAdeno-HSV-2 gD-DC可以诱导小鼠脾淋巴细胞增殖（刺激指数为1.600±0.215）、有效杀伤靶细胞（CTL活性为37.1％）,与pAdeno-DC组（分别为1.063±0.070和16.0％）、DC组（1.056±0.063和14.9％）、空白对照组（1.020±0.051和15.7％）比较,差异均有统计学意义（P＜0.05）.pAdeno-HSV-2 gD-DC组小鼠分离的脾细胞培养上清可以检测到高水平IFN-γ（A450值为0.568±0.031）和IL-4（A450值为0.544±0.043）,与pAdeno-DC组（0.266±0.021和0.278±0.037）、DC组（0.271±0.023和0.275±0.044）、空白对照组（0.252±0.012和0.245±0.051）比较,差异均有统计学意义（P＜0.05）.结论 构建的pAdeno-HSV-2 gD-DC疫苗可以在小鼠中产生较强的特异性免疫应答.Objective To evaluate the specific immune response induced by a dendritic cell-based adenovirus-mediated vaccine carrying the herpes simplex virus type 2 glycoprotein D gene （pAdeno-HSV-2 gD-DC） in BALB/c mice.Methods Forty BALB/c mice were equally divided into four groups：blank control group receiving no treatment,pAdeno-DC group immunized with pAdeno-DC,pAdeno-HSV-2 gD-DC group immunized with the previously constructed vaccine pAdeno-HSV-2 gD-DC,DC group immunized with DCs only.Totally,three rounds of vaccination were conducted at a 7-day interval.Ten days after the last vaccination,serum samples were collected and spleen cells were isolated from these mice.Enzyme-linked immunosorbent assay （ELISA） was performed to measure the level of IgG antibody against HSV-2 gD in the serum samples.Some spleen cells were stimulated with HSV-2 gD protein （10 mg/L） for 72 hours; then,ELISA was carried out to determine the levels of interferon （IFN）-γand interleukin （IL）-4 in the supernatant,and 3-（4,5-dimethylthiazol-2-yl）-2,5 diphenyl tetrazolium bromide （MTT） assay to estimate the proliferative activity of these cells.The cytotoxicity of spleen cells was also evaluated based on the measurement of lactate dehydrogenase （LDH） release.Results The serum level of IgG antibody against HSV-2 gD （given in the absorbance value at 450 nm） was 0.313 ± 0.034 in the pAdeno-HSV-2 gD-DC group,significantly higher than that in the pAdeno-DC group,DC group and blank control group （0.034 ± 0.009,0.028 ± 0.009 and 0.026 ± 0.010 respectively,all P ＜ 0.05）.Increased proliferative activity and cytotoxicity were observed in spleen cells from the pAdeno-HSV-2 gD-DC group compared with those from the pAdeno-DC group,DC group and blank control group （cell stimulation index：1.600 ± 0.215 vs.1.063 ± 0.070,1.056 ± 0.063 and 1.020 ± 0.051,all P ＜ 0.05; percentage of cytotoxicity：37.1％ vs.16.0％,14.9％ and 15.7％,all P ＜ 0.05）.The levels of IFN-γ and IL-4 （both given in the absorban

关 键 词：疱疹病毒2型 人 树突细胞 单纯疱疹病毒疫苗 

分 类 号：R392[医药卫生—免疫学]