Lovett A E, McCarthy M, Wolinsky J S
University of Texas Health Science Center, Department of Neurology, Houston 77030.
J Gen Virol. 1993 Mar;74 ( Pt 3):445-52. doi: 10.1099/0022-1317-74-3-445.
Although it is known that rubella-immune individuals have T cells that proliferate in vitro in response to rubella virus (RV), the determinants that evoke this response have not been identified. This study utilized recombinant proteins that express overlapping sequences of the RV structural open reading frame to identify domains of the structural proteins that contain cell-mediated immunodominant sequences. Lysates enriched with RV fusion proteins (RecA-RV-LacZ) were prepared from Escherichia coli transformed with plasmids which contained specific RV cDNA inserts. Approximately 62% of RV-immune individuals gave RV-specific responses to one or more of the RV fusion proteins. Over 10% of immune individuals recognized the capsid sequence C1-C29. Lymphoproliferation data from studies using six overlapping synthetic peptides representing this sequence suggested that as much as 70% of the immune population may recognize this domain. An E1 sequence, E1(202)-E1(283), was recognized by 15% of the RV-immune individuals with the fusion proteins. Five synthetic peptides representing this sequence had an overall response rate of 50%. The sequence C64-C97 failed to evoke any RV-specific responses with the fusion proteins and synthetic peptides representing this sequence were used to verify that the RV fusion proteins and the criteria used to identify RV-specific responses were adequate. These peptides gave a response rate of only 6%. In general, significant responses to specific fusion proteins correlated with high responses (stimulation index > or = 4.0) to representative synthetic peptides. This study suggests that the recombinant proteins were beneficial in identifying cell-mediated immunodominant domains of the RV structural proteins which could be further characterized with synthetic peptides.
尽管已知风疹免疫个体具有可在体外对风疹病毒（RV）产生增殖反应的T细胞，但引发这种反应的决定因素尚未确定。本研究利用表达RV结构开放阅读框重叠序列的重组蛋白，来鉴定结构蛋白中包含细胞介导免疫显性序列的结构域。从用含有特定RV cDNA插入片段的质粒转化的大肠杆菌中制备富含RV融合蛋白（RecA-RV-LacZ）的裂解物。约62%的风疹免疫个体对一种或多种RV融合蛋白产生风疹特异性反应。超过10%的免疫个体识别衣壳序列C1-C29。使用代表该序列的六种重叠合成肽进行研究所得的淋巴细胞增殖数据表明，多达70%的免疫人群可能识别该结构域。15%的风疹免疫个体在用融合蛋白检测时识别E1序列E1(202)-E1(283)。代表该序列的五种合成肽的总体反应率为50%。序列C64-C97在用融合蛋白检测时未引发任何风疹特异性反应，并且使用代表该序列的合成肽来验证RV融合蛋白及用于鉴定风疹特异性反应的标准是合适的。这些肽的反应率仅为6%。一般来说，对特定融合蛋白的显著反应与对代表性合成肽的高反应（刺激指数≥4.0）相关。本研究表明，重组蛋白有助于鉴定RV结构蛋白的细胞介导免疫显性结构域，这些结构域可用合成肽作进一步表征。
