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首页> 美国卫生研究院文献>The Journal of Experimental Medicine >Processing of a Multiple Membrane Spanning Epstein-Barr Virus Protein for Cd8+T Cell Recognition Reveals a Proteasome-Dependent Transporter Associated with Antigen Processing–Independent Pathway
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Processing of a Multiple Membrane Spanning Epstein-Barr Virus Protein for Cd8+T Cell Recognition Reveals a Proteasome-Dependent Transporter Associated with Antigen Processing–Independent Pathway

机译：处理跨膜的爱泼斯坦-巴尔病毒蛋白的Cd8 + T细胞识别过程揭示了蛋白酶体依赖转运蛋白与抗原加工独立途径有关。

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摘要

Epstein-Barr virus (EBV) latent membrane protein (LMP)2 is a multiple membrane spanning molecule which lacks ectodomains projecting into the lumen of the endoplasmic reticulum (ER). Human CD8+ cytotoxic T lymphocytes (CTL)s recognize a number of epitopes within LMP2. Assays with epitope-specific CTLs in two different cell backgrounds lacking the transporter associated with antigen processing (TAP) consistently show that some, but not all, LMP2 epitopes are presented in a TAP-independent manner. However, unlike published examples of TAP-independent processing from endogenously expressed antigens, presentation of TAP-independent LMP2 epitopes was abrogated by inhibition of proteasomal activity. We found a clear correlation between hydrophobicity of the LMP2 epitope sequence and TAP independence, and experiments with vaccinia minigene constructs expressing cytosolic epitope peptides confirmed that these more hydrophobic peptides were selectively able to access the HLA class I pathway in TAP-negative cells. Furthermore, the TAP-independent phenotype of particular epitope sequences did not require membrane location of the source antigen since (i) TAP-independent LMP2 epitopes inserted into an EBV nuclear antigen and (ii) hydrophobic epitope sequences native to EBV nuclear antigens were both presented in TAP-negative cells. We infer that there is a proteasome-dependent, TAP-independent pathway of antigen presentation which hydrophobic epitopes can selectively access.
机译：爱泼斯坦-巴尔病毒（EBV）潜在膜蛋白（LMP）2是跨膜的多分子分子，缺少投射到内质网（ER）内腔的胞外域。人类CD8 + 细胞毒性T淋巴细胞（CTL）识别LMP2中的许多表位。在缺少抗原处理（TAP）相关转运蛋白的两种不同细胞背景中，针对抗原决定簇特异性CTL的测定始终显示出，但不是全部，LMP2抗原决定簇以不依赖TAP的方式出现。但是，与已发表的内源性表达抗原不依赖TAP的加工方法不同，不依赖TAP的LMP2表位的表达被蛋白酶体活性抑制所废除。我们发现LMP2表位序列的疏水性与TAP独立性之间存在明显的相关性，并且使用表达细胞溶质表位肽的牛痘小基因构建体进行的实验证实，这些更具疏水性的肽能够选择性地进入TAP阴性细胞中的HLA I类途径。此外，特定表位序列的不依赖TAP的表型不需要源抗原的膜位置，因为（i）插入了EBV核抗原的不依赖TAP的LMP2表位和（ii）EBV核抗原固有的疏水性表位序列都被提出在TAP阴性细胞中。我们推断，抗原呈递存在蛋白酶体依赖性，TAP依赖性途径，疏水性抗原决定簇可以选择性进入。

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