Malkinson M, Davidson I, Becker Y
Department of Avian Diseases, Kimron Veterinary Institute, Bet Dagan, Israel.
Arch Virol. 1992;127(1-4):169-84. doi: 10.1007/BF01309582.
Antigen B of Marek's disease virus (MDV) vaccine strains CVI988 and SB1 (serotypes 1 and 2) and herpesvirus of turkeys (HVT) (serotype 3) is formed of oligomeric molecules that are detergent-stable and heat-labile. Immunoblots of native membranal extracts of HVT- and MDV-infected chick embryo fibroblasts (CEF) probed with avian monoserotypic antisera, murine monoclonal antibodies (mAb) to the three serotypes and mAb to antigen B showed two distinct patterns of high molecular weight oligomeric antigens. Serotypes 1 and 3 vaccine viruses formed one set and serotype 2, the other. Avian monotypic sera to serotypes 1 and 3 viruses detected two high molecular weight bands of 230 and > or = 300 kDa in MDV-1 and HVT-infected CEF but only a weak diffuse zone ranging from 130 to 230 kDa in extracts of SB1-infected CEF. No 300 kDa band was discernible in the SB1 extract when blotted with avian monotypic 1 and 3 antisera. MAbs to MDV serotypes 1 and 3 and to antigen B also detected the 230 and > or = 300 kDa antigens, while the mAb to SB1 detected a 50 kDa antigen in the SB1-infected extract only. Furthermore, the antigen B mAb did not reveal high mol. wt. oligomers in SB1-infected CEF extracts. Antigen B oligomers were rapidly destroyed by heating at 95 degrees C and the rate of denaturation of the 230 and > or = 300 kDa oligomers differed for each of the three vaccine viruses. We propose that antigen B of MDV1 and HVT has a complex conformation created by juxtaposition of dimers (230-250 kDa) and trimers (> or = 300 kDa), and is inserted in the infected cell membrane so that conformational, discontinuous epitopes are formed in addition to continuous epitopes. It appears that HVT protects chickens against oncogenic strains of MDV1 by virtue of the cross reactivity of the conformational determinants located on these oligomers. Serotype 2 vaccine shares some of its antigenic determinants with serotypes 1 and 3, while its unique immunogenic features form the basis of the protective synergism achieved when serotypes 2 and 3 vaccines are combined together.
马立克氏病病毒(MDV)疫苗株CVI988和SB1(血清型1和2)以及火鸡疱疹病毒(HVT)(血清型3)的抗原B由对去污剂稳定且对热不稳定的寡聚分子组成。用禽单血清型抗血清、针对这三种血清型的鼠单克隆抗体(mAb)以及针对抗原B的mAb对感染HVT和MDV的鸡胚成纤维细胞(CEF)的天然膜提取物进行免疫印迹,显示出两种不同模式的高分子量寡聚抗原。血清型1和3疫苗病毒形成一组,血清型2形成另一组。针对血清型1和3病毒的禽单型血清在感染MDV - 1和HVT的CEF中检测到两条高分子量条带,分别为230 kDa和≥300 kDa,但在感染SB1的CEF提取物中仅检测到一个从130 kDa到230 kDa的弱弥散区。用禽单型1和3抗血清印迹时,在SB1提取物中未观察到300 kDa的条带。针对MDV血清型1和3以及抗原B的mAb也检测到230 kDa和≥300 kDa的抗原,而针对SB1的mAb仅在感染SB1的提取物中检测到一个50 kDa的抗原。此外,抗原B的mAb在感染SB1的CEF提取物中未显示高分子量寡聚物。抗原B寡聚物在95℃加热时迅速被破坏,并且三种疫苗病毒各自的230 kDa和≥300 kDa寡聚物的变性速率不同。我们提出,MDV1和HVT的抗原B具有由二聚体(230 - 250 kDa)和三聚体(≥300 kDa)并列形成的复杂构象,并插入感染细胞膜中,从而除了连续表位之外还形成构象性、不连续表位。似乎HVT凭借这些寡聚物上构象决定簇的交叉反应性保护鸡免受MDV1致癌株的侵害。血清型2疫苗与血清型1和3共享一些抗原决定簇,而其独特的免疫原性特征构成了血清型2和3疫苗联合使用时实现保护协同作用的基础。