van Rijn P A, van Gennip H G, Moormann R J
Department of Mammalian Virology, Institute for Animal Science and Health, Lelystad, The Netherlands.
Vaccine. 1999 Feb 5;17(5):433-40. doi: 10.1016/s0264-410x(98)00215-1.
Envelope glycoprotein E2 is the most immunogenic protein of classical swine fever virus (CSFV). In a proposed model of the antigenic structure of E2, the N-terminal half of E2 forms two independent structural antigenic units, A and BC. E2 without transmembrane region (E2-TMR) is expressed and secreted into the medium of insect cells by use of the baculovirus expression system. The immune response induced by E2 protects pigs against CSFV. Recently, we showed that the protective immune response to a homologous CSFV challenge can be induced by a single unit, A or BC, of E2. An indirect blocking ELISA, or complex trapping blocking assay (CTB) based on both units is routinely used worldwide for serological diagnosis of CSFV infections. Here we show that E2-TMR is secreted into the medium as a homodimer. This E2 homodimer was used to develop a CTB detecting antibodies directed against one immunogenic unit of E2. Thus, the protective immune response induced by E2 containing one unit was not detected with a modified CTB based on the other unit, whereas immune responses induced by a variety of low virulent CSFV strains were detected with such a modified CTB. These results indicate that a deletion E2 protein in combination with a modified CTB are feasible as CSF marker vaccine and accompanying differentiating diagnostic test.
包膜糖蛋白E2是经典猪瘟病毒(CSFV)最具免疫原性的蛋白。在一个提出的E2抗原结构模型中,E2的N端半段形成两个独立的结构抗原单位,A和BC。通过杆状病毒表达系统表达无跨膜区的E2(E2-TMR)并分泌到昆虫细胞培养基中。E2诱导的免疫反应可保护猪抵抗CSFV。最近,我们发现E2的单个单位A或BC可诱导针对同源CSFV攻击的保护性免疫反应。基于这两个单位的间接阻断ELISA或复合捕获阻断试验(CTB)在全球范围内常规用于CSFV感染的血清学诊断。在此我们表明E2-TMR以同二聚体形式分泌到培养基中。这种E2同二聚体用于开发一种CTB,以检测针对E2一个免疫原性单位的抗体。因此,基于另一个单位的改良CTB未检测到由含一个单位的E2诱导的保护性免疫反应,而用这种改良CTB检测到了由多种低毒力CSFV毒株诱导的免疫反应。这些结果表明,缺失E2蛋白与改良CTB相结合作为CSF标记疫苗和配套鉴别诊断试验是可行的。