Reay P A, Jones I M, Brownlee G G
Sir William Dunn School of Pathology, University of Oxford, United Kingdom.
Virology. 1988 Nov;167(1):261-8. doi: 10.1016/0042-6822(88)90076-1.
We have used a recombinant vaccinia virus to investigate the recognition of the PB2 protein of influenza A/NT/60/68 (H3N2) by murine polyclonal CTL populations. PB2 is recognized as a major cross-reactive target antigen. Recognition of PB2 is under strict genetic control, since BALB/c (H-2d) but not CBA (H-2k) mice are responders. We also demonstrate, by use of cell lines transfected with individual genes encoding class I molecules of the H-2d haplotype, that recognition of PB2 occurs in conjunction with the H-2Dd but not the H-2Kd or H-2Ld molecules. In contrast, recognition of the nucleoprotein of A/PR/8/34 by BALB/c-derived polyclonal CTL is restricted via the H-2Kd molecule. By using three recombinant vaccinia viruses expressing deleted forms of the PB2 protein we show that at least one epitope of the PB2 protein resides within the amino-terminal 256 amino acids. This approach offers an effective method to map the regions of large proteins containing epitopes recognized by CTL.
我们利用重组痘苗病毒来研究甲型流感病毒A/NT/60/68（H3N2）的PB2蛋白被鼠多克隆CTL群体识别的情况。PB2被认为是一种主要的交叉反应靶抗原。PB2的识别受到严格的基因控制，因为BALB/c（H-2d）小鼠是反应者，而CBA（H-2k）小鼠则不是。我们还通过使用转染了编码H-2d单倍型I类分子的单个基因的细胞系证明，PB2的识别与H-2Dd分子相关，而与H-2Kd或H-2Ld分子无关。相比之下，BALB/c来源的多克隆CTL对A/PR/8/34核蛋白的识别是通过H-2Kd分子受限的。通过使用三种表达PB2蛋白缺失形式的重组痘苗病毒，我们表明PB2蛋白的至少一个表位位于氨基末端的256个氨基酸内。这种方法为绘制包含被CTL识别的表位的大蛋白区域提供了一种有效的方法。
