Karem K L, Bowen J, Kuklin N, Rouse B T
Department of Microbiology and Immunology, College of Veterinary Medicine, University of Tennessee at Knoxville, 37996, USA.
J Gen Virol. 1997 Feb;78 ( Pt 2):427-34. doi: 10.1099/0022-1317-78-2-427.
Salmonella typhimurium strains expressing foreign antigens of various pathogens are capable of eliciting antigen-specific humoral and cellular immune responses. Attenuated S. typhimurium strain chi4550 (delta(cya) delta(crp) delta(asd)) was used as an expression vector for herpes simplex virus (HSV) antigens. Genes encoding glycoprotein D (gD) and the immediate-early protein ICP27 of HSV-1 were cloned and expressed in plasmid pYA292 (asd+) and subsequently placed into chi4550. Following two oral immunizations, the protective efficacy of recombinant strains against zosteriform challenge with HSV-1 was measured in 3-4-week-old BALB/c mice. Levels of protection observed were 77% with the ICP27 construct but only 31% with the gD construct. Zosteriform protection correlates with a CD4+-mediated delayed-type hypersensitivity (DTH) reaction against HSV. Accordingly, significant DTH was observed only in mice immunized orally with the S. typhimurium ICP27 construct. ELISA analysis of antigen-specific humoral responses failed to detect serum antibody responses following oral administration although recombinant S. typhimurium were isolated from spleens of orally dosed mice up to day 30. Intravenous (i.v.) immunization with the gD-expressing construct did, however, induce detectable serum antibody responses. Some humoral IgA responses against gD in faecal samples were detected as early as 3 weeks post-oral immunization while those induced by the i.v. route were slightly lower. These data suggest that recombinant S. typhimurium HSV antigens are capable of inducing immunity against HSV, some aspects of which are protective against HSV challenge.
表达各种病原体外源抗原的鼠伤寒沙门氏菌菌株能够引发抗原特异性体液免疫和细胞免疫反应。减毒鼠伤寒沙门氏菌菌株chi4550（Δ（cya）Δ（crp）Δ（asd））被用作单纯疱疹病毒（HSV）抗原的表达载体。编码HSV-1糖蛋白D（gD）和立即早期蛋白ICP27的基因被克隆并在质粒pYA292（asd+）中表达，随后导入chi4550。经过两次口服免疫后，在3至4周龄的BALB/c小鼠中测定重组菌株对HSV-1带状疱疹样攻击的保护效力。观察到的保护水平，ICP27构建体为77%，而gD构建体仅为31%。带状疱疹样保护与针对HSV的CD4+介导的迟发型超敏反应（DTH）相关。因此，仅在口服鼠伤寒沙门氏菌ICP27构建体免疫的小鼠中观察到显著的DTH。抗原特异性体液反应的ELISA分析未能检测到口服给药后的血清抗体反应，尽管在口服给药小鼠的脾脏中直到第30天仍可分离到重组鼠伤寒沙门氏菌。然而，用表达gD的构建体进行静脉内（i.v.）免疫确实诱导了可检测的血清抗体反应。早在口服免疫后3周就检测到粪便样本中针对gD的一些体液IgA反应，而静脉内途径诱导的反应略低。这些数据表明，重组鼠伤寒沙门氏菌HSV抗原能够诱导针对HSV的免疫，其中一些方面对HSV攻击具有保护作用。
