Pan Mingjie, Wang Xingsheng, Liao Jianmin, Yin Dengke, Li Suqin, Pan Ying, Wang Yao, Xie Guangyan, Zhang Shumin, Li Yuexi
Department of Biochemistry and Molecular Biology, School of Preclinical Medicine, Nanjing Medical University, Nanjing 210029, China.
Clin Dev Immunol. 2012;2012:205313. doi: 10.1155/2012/205313. Epub 2012 May 9.
Twenty B candidate epitopes of glycoproteins B (gB2), C (gC2), E (gE2), G (gG2), and I (gI2) of herpes simplex virus type 2 (HSV-2) were predicted using DNAstar, Biosun, and Antheprot methods combined with the polynomial method. Subsequently, the biological functions of the peptides were tested via experiments in vitro. Among the 20 epitope peptides, 17 could react with the antisera to the corresponding parent proteins in the EIA tests. In particular, five peptides, namely, gB2(466-473) (EQDRKPRN), gC2(216-223) (GRTDRPSA), gE2(483-491) (DPPERPDSP), gG2(572-579) (EPPDDDDS), and gI2(286-295) (CRRRYRRPRG) had strong reaction with the antisera. All conjugates of the five peptides with the carrier protein BSA could stimulate mice into producing antibodies. The antisera to these peptides reacted strongly with the corresponding parent glycoproteins during the Western Blot tests, and the peptides reacted strongly with the antibodies against the parent glycoproteins during the EIA tests. The antisera against the five peptides could neutralize HSV-2 infection in vitro, which has not been reported until now. These results suggest that the immunodominant epitopes screened using software algorithms may be used for virus diagnosis and vaccine design against HSV-2.
运用DNAstar、Biosun和Antheprot方法并结合多项式方法,预测了2型单纯疱疹病毒(HSV-2)糖蛋白B(gB2)、C(gC2)、E(gE2)、G(gG2)和I(gI2)的20个B类候选表位。随后,通过体外实验对这些肽段的生物学功能进行了检测。在这20个表位肽中,有17个在酶免疫分析(EIA)试验中能与相应亲本蛋白的抗血清发生反应。特别地,5个肽段,即gB2(466 - 473)(EQDRKPRN)、gC2(216 - 223)(GRTDRPSA)、gE2(483 - 491)(DPPERPDSP)、gG2(572 - 579)(EPPDDDDS)和gI2(286 - 295)(CRRRYRRPRG)与抗血清有强烈反应。这5个肽段与载体蛋白牛血清白蛋白(BSA)的所有偶联物均可刺激小鼠产生抗体。在蛋白质印迹(Western Blot)试验中,这些肽段的抗血清与相应的亲本糖蛋白发生强烈反应,而在EIA试验中,这些肽段与亲本糖蛋白的抗体也发生强烈反应。针对这5个肽段的抗血清在体外可中和HSV-2感染,这一结果此前未见报道。这些结果表明,利用软件算法筛选出的免疫显性表位可用于HSV-2的病毒诊断和疫苗设计。