贾岚;傅婷;陶薇;何卓晶;洪艳;

• 1:浙江省医学科学院生物工程研究所

摘要(Abstract)：

目的研究霍乱病毒B亚单位(cholera toxin subunit B,CTB)作为基因佐剂对单纯疱疹病毒2型(herpes simplex virus type 2,HSV-2)DNA疫苗pg D免疫效果的影响。方法将CTB片段克隆至真核表达质粒pc DNA3Kan(p Kan)中,构建基因佐剂pc DNA3Kan-CTB(p CTB)。将BALB/c小鼠随机分为p CTB无疫苗组、pg D/p CTB佐剂组、pg D/p Kan空载体组及PBS对照组,经后腿肌肉多点注射,共免疫3次,每次间隔2周。于末次免疫2周后,经小鼠眼球采血,分离血清,ELISA法检测HSV-2特异性Ig G水平及Ig G1、Ig G2a亚型抗体滴度;制备小鼠脾细胞,MTS法检测小鼠脾脏T细胞增殖能力;将病毒稀释后经腹腔接种小鼠,于免疫2周后,进行HSV-2致死剂量攻毒试验。结果p CTB经双酶切及测序鉴定证明构建正确;pg D/p CTB佐剂组小鼠血清中HSV-2特异性Ig G水平明显高于其他3组(P<0.001),Ig G1和Ig G2a滴度均明显高于pg D/p Kan空载体组(P<0.001),其中Ig G2a增幅更明显,Ig G2a/Ig G1比值与pg D/p Kan空载体组相比,由(1.176±0.11)增加至(1.316±0.07)(P<0.05);pg D/p CTB佐剂组小鼠脾细胞在HSV-2刺激下诱导的特异性T细胞增殖反应明显强于PBS对照组及pg D/p Kan空载体组(P<0.05);攻毒14 d内,pg D/p CTB佐剂组小鼠存活率高达100%。结论 p CTB作为基因佐剂能够显著提高pg D诱导小鼠产生抗原特异性Ig G抗体水平,增强HSV-2特异性T细胞增殖反应,并介导以细胞免疫为主的Th1/Th2混合型免疫反应。

关键词(KeyWords)： 霍乱病毒B亚单位;单纯疱疹病毒2型(HSV-2);DNA疫苗;基因佐剂

Abstract:

Keywords:

基金项目(Foundation): 浙江省自然科学基金(LY12H19009);; 浙江省科技厅计划项目(2011C23002);浙江省科技厅计划项目(2011F20015)

作者(Author): 贾岚;傅婷;陶薇;何卓晶;洪艳;

Email:

DOI: 10.13200/j.cnki.cjb.001316

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