中华皮肤科杂志 ›› 2001, Vol. 30 ›› Issue (1): 53-54.

• 论著 • 上一篇    下一篇

疱疹病毒gD基因多聚核苷酸疫苗的构建和表达

杨慧兰1, 廖元兴1, 杨静2, 王捷2, 杨太成2, 田野3, 刘荣卿4   

  1. 1. 广州军区总医院皮肤科, 广州 510010;
    2. 广州军区总医院医学实验科;
    3. 广州军区总医院病理科;
    4. 第三军医大学西南医院皮肤科
  • 收稿日期:2000-02-24 出版日期:2001-02-15 发布日期:2001-02-15
  • 基金资助:
    广东省科委自然科学基金资助(960671)

Construction and Expression of Herpes Simplex Virus Glycoprotein Polynucleotide Vaccine

YANG Huilan1, YANG Jing1, LIAO Yuanxing2   

  1. Guangzhou General Hospital, Guangzhou Military Command, Guangzhou 510010
  • Received:2000-02-24 Online:2001-02-15 Published:2001-02-15

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摘要： 目的 构建疱疹病毒gD2基因多聚核苷酸疫苗并在实验动物中得到表达.方法 将构建的pcDNA3-gD2的质粒DNA注射入豚鼠的舌肌中,每周1次,共3次,最后1次免疫48h后引颈处死豚鼠,并取其舌、肝、脾、肾、心脏及股四头肌,用免疫组化法(SP法)检测gD2抗原的表达.结果 重组质粒组gD2真核表达载体免疫接种后,均可在舌肌测到抗原表达,而对照组均为阴性.结论 重组质粒gD2真核表达载体可以在豚鼠舌肌中表达,对建立基因免疫防治疱疹病毒感染奠定一定基础.

关键词: 疱疹,生殖器, 糖蛋白D, 多聚核苷酸疫苗

Abstract: Objective To construct Herpes simplex virus-2 glycoprotein D polynucleotide vaccine,and to detect the glycoprotein D2(gD2)expression in guinea pig's tongue by intramuscular injection of gD2 polynucleotide vaccine. Methods The recombinant plasmids HSV-gD2 polynucleotide vaccine was constructed by molecular cloning technique. The guniea pigs were intramuscularly injected with 0.75% bupivacaine,after 7 days,they were divided into 3 groups(4 guinea pigs per group)and intramuscularly injected with 100μL 0.09% NaCl、pcDNA3、gD2-pcDNA3,three times at 7 day intervals respectively. All guinea pigs were sacrificed 2 days after the last injection,and the tongue,liver,kidney,spleen and quadricep muscle were removed for immunohistochemical assay (SP method). Results The recombinant pcDNA3-gD2 plasmid was constructed succuessfully. Immunohistochemical staining for gD2 was positive in the tongue injected with pcDNA3-gD2,but negative in the liver,kidney,spleen,heart and quadricep muscle. Conclusion Herpes simplex virus-2 glycoprotein D polynucleotide vaccine was constructed successfully,and gD2 antigen can be expressed effectively in the tongue after injection of the HSV-gD2 polynucleotide vaccine in it.

Key words: Herpes,genitalis, Glycoprotein D, Polynucleotide vaccines