Schleiss M R, Bourne N, Jensen N J, Bravo F, Bernstein D I
Division of Infectious Diseases, Children's Hospital Research Foundation, Cincinnati, Ohio 45229, USA.
Viral Immunol. 2000;13(2):155-67. doi: 10.1089/vim.2000.13.155.
Vaccines are needed for control of congenital human cytomegalovirus (HCMV) infection. Although the species-specificity of cytomegaloviruses precludes preclinical evaluation of HCMV vaccines in animal models, the guinea pig cytomegalovirus (GPCMV), which causes disease in utero, is a relevant model for the study of vaccines against congenital infection. We investigated whether DNA vaccines that target two GPCMV proteins, glycoprotein B (gB) and UL83 (pp65), are capable of eliciting immune responses in vivo. After cloning each gene into an expression vector, DNA was delivered by intramuscular inoculation and by pneumatic epidermal delivery. In Swiss-Webster mice, anti-gB titers were significantly higher after epidermal delivery. After epidermal inoculation in guinea pigs, all gB-immunized animals (n = 6) had antibody responses comparable to those induced by natural infection. Viral neutralization titers ranged from 1:64 to greater than 1:128. A GPCMV UL83 DNA vaccine also elicited an antibody response in all immunized guinea pigs (n = 6) after epidermal administration. Immunoprecipitation and Western blot assays confirmed that immune sera were immunoreactive with virion-associated UL83 and gB proteins. We conclude that DNA vaccines against GPCMV structural proteins are immunogenic, and warrant further investigation in the guinea pig model of congenital CMV infection.
控制先天性人类巨细胞病毒(HCMV)感染需要疫苗。尽管巨细胞病毒的种属特异性使得无法在动物模型中对HCMV疫苗进行临床前评估,但豚鼠巨细胞病毒(GPCMV)可在子宫内引发疾病,是研究先天性感染疫苗的相关模型。我们研究了针对两种GPCMV蛋白,即糖蛋白B(gB)和UL83(pp65)的DNA疫苗是否能够在体内引发免疫反应。将每个基因克隆到表达载体中后,通过肌肉注射和气力表皮给药方式递送DNA。在瑞士韦伯斯特小鼠中,表皮给药后抗gB滴度显著更高。在豚鼠中进行表皮接种后,所有接种gB的动物(n = 6)产生的抗体反应与自然感染诱导的反应相当。病毒中和滴度范围为1:64至大于1:128。在表皮给药后,GPCMV UL83 DNA疫苗也在所有接种的豚鼠(n = 6)中引发了抗体反应。免疫沉淀和蛋白质印迹分析证实,免疫血清与病毒体相关的UL83和gB蛋白具有免疫反应性。我们得出结论,针对GPCMV结构蛋白的DNA疫苗具有免疫原性,值得在先天性CMV感染的豚鼠模型中进一步研究。