Discovery of CD8+ T cell epitopes in Chlamydia trachomatis infection through use of caged class I MHC tetramers
Discovery of CD8+ T cell epitopes in Chlamydia trachomatis infection through use of caged class I MHC tetramers
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DOI：
10.1073/pnas.0711504105
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发表时间：
2008-03-11
影响因子：
11.1
通讯作者：
Ploegh, Hidde L.
中科院分区：
文献类型：
--
作者：
Grotenbreg, Gijsbert M.;Roan, Nadia R.;Ploegh, Hidde L.
一类MHC四聚体允许CD8(+) T细胞群的直接表型鉴定，但它们的生产仍然很费力。利用装载条件配体（笼化MHC分子）的I类MHC产物的肽交换策略提供了一种快速直接的方法来获得不同的I类MHC四聚体阵列，并促进CD8(+) T细胞表位的发现。在这里，我们描述了FAPGNYPAL （SV9）表位的光可切割类似物的发展，这些类似物结合H-2K(b)和H-2D(b)，并完全保留其结构和功能完整性。我们对沙眼衣原体基因组中所有可能的H-2Kb八聚体和H-2D(b)非美聚体表位进行了排序，并用选择的肽替代SV9类似物，从大约2000个得分最高的肽中制备了MHC四聚体。由此产生的2000个成员的I类MHC四聚体阵列允许发现来自多态膜蛋白I （PmpI）的表位的两个变体，并评估相应CD8(+) T细胞的涌现动力学和效应功能。
Class I MHC tetramers allow direct phenotypic identification of CD8(+) T cell populations, but their production remains laborious. A peptide exchange strategy that employs class I MHC products loaded with conditional ligands (caged MHC molecules) provides a fast and straightforward method to obtain diverse arrays of class I MHC tetramers and facilitates CD8(+) T cell epitope discovery. Here, we describe the development of photocleavable analogs of the FAPGNYPAL (SV9) epitope that bind H-2K(b) and H-2D(b) with full retention of their structural and functional integrity. We ranked all possible H-2Kb octameric and H-2D(b) nonameric epitopes that span the genome of Chlamydia trachomatis and prepared MHC tetramers from approximate to 2,000 of the highest scoring peptides by replacement of the SV9 analog with the peptide of choice. The resulting 2,000-member class I MHC tetramer array allowed the discovery of two variants of an epitope derived from polymorphic membrane protein I (PmpI) and an assessment of the kinetics of emergence and the effector function of the corresponding CD8(+) T cells.