Harder T C, Harder M, de Swart R L, Osterhaus A D, Liess B
Department of Virology, Erasmus University, Rotterdam, The Netherlands.
Vet Q. 1998 Apr;20(2):50-5. doi: 10.1080/01652176.1998.9694838.
The immunogenic proteins of cells infected with the alpha- or the gamma-herpesvirus of seals, phocid herpesvirus-1 and -2 (PhHV-1, -2), were examined in radioimmunoprecipitation assays as a further step towards the development of a PhHV-1 vaccine. With sera obtained from convalescent seals of different species or murine monoclonal antibodies (Mabs), at least seven virus-induced glycoproteins were detected in lysates of PhHV-1-infected CrFK cells. A presumably disulphide-linked complex composed of glycoproteins of 59, 67 and 113/120 kDa, expressed on the surface of infected cells, was characterized as a major immunogenic infected cell protein of PhHV-1. This glycoprotein complex has previously been identified as the proteolytically cleavable glycoprotein B homologue of PhHV-1 (14). At least three distinct neutralization-relevant epitopes were operationally mapped, by using Mabs, on the glycoprotein B of PhHV-1. Among the infected cell proteins of the antigenically closely related feline and canine herpesvirus, the glycoprotein B equivalent proved to be the most highly conserved glycoprotein. Sera obtained from different seal species from Arctic, Antarctic, and European habitats did not precipitate uniform patterns of infected cell proteins from PhHV-1-infected cell lysates although similar titres of neutralizing antibodies were displayed. Thus, antigenic differences among the alphaherpesvirus species prevalent in the different pinniped populations cannot be excluded. PhHV-2 displayed a different pattern of infected cell proteins and only limited cross-reactivity to PhHV-1 at the protein level was detected, which is in line with its previous classification as a distinct species, based on nucleotide sequence analysis, of the gammaherpesvirus linenge. A Mab raised against PhHV-2 and specific for a major glycoprotein of 117 kDa, cross reacted with the glycoprotein B of PhHV-1. The 117-kDa glycoprotein could represent the uncleaved PhHV-2 glycoprotein B homologue.
为了进一步研发海豹1型疱疹病毒疫苗，我们通过放射免疫沉淀试验检测了感染海豹α或γ疱疹病毒（海豹疱疹病毒1型和2型，即PhHV-1和PhHV-2）的细胞的免疫原性蛋白。使用从不同物种的康复海豹获得的血清或鼠单克隆抗体（Mab），在感染PhHV-1的CrFK细胞裂解物中检测到至少七种病毒诱导的糖蛋白。一种可能由59、67和113/120 kDa糖蛋白组成的二硫键连接复合物，在感染细胞表面表达，被鉴定为PhHV-1的主要免疫原性感染细胞蛋白。这种糖蛋白复合物先前已被鉴定为PhHV-1的可被蛋白酶切割的糖蛋白B同源物（14）。通过使用Mab，在PhHV-1的糖蛋白B上至少定位了三个不同的与中和相关的表位。在抗原性密切相关的猫和犬疱疹病毒的感染细胞蛋白中，糖蛋白B等效物被证明是最保守的糖蛋白。尽管显示出相似滴度的中和抗体，但从北极、南极和欧洲栖息地的不同海豹物种获得的血清并不能沉淀出PhHV-1感染细胞裂解物中感染细胞蛋白的统一模式。因此，不能排除不同鳍足类动物群体中普遍存在的α疱疹病毒物种之间的抗原差异。PhHV-2显示出不同的感染细胞蛋白模式，并且在蛋白质水平上仅检测到与PhHV-1的有限交叉反应，这与其先前基于核苷酸序列分析被分类为γ疱疹病毒谱系中的一个独特物种一致。一种针对PhHV-2产生的、对117 kDa主要糖蛋白具有特异性的Mab与PhHV-1的糖蛋白B发生交叉反应。117 kDa糖蛋白可能代表未切割的PhHV-2糖蛋白B同源物。
