Rapid selection and identification of functional CD8+ T cell epitopes from large peptide-coding libraries
Rapid selection and identification of functional CD8+ T cell epitopes from large peptide-coding libraries
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DOI：
10.1038/s41467-019-12444-7
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发表时间：
2019-10
影响因子：
16.6
通讯作者：
Govinda Sharma;C. Rive;R. Holt
中科院分区：
文献类型：
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作者：
Govinda Sharma;C. Rive;R. Holt
细胞毒性CD 8 +T细胞识别并消除在其表面上呈递源自细胞内加工抗原的肽表位的感染细胞或恶性细胞。然而，全面剖析特定的主要组织相容性复合体（MHC）结合的肽表位，自然加工，并能够引发功能性T细胞反应一直具有挑战性。在这里，我们报告了一种用于深度和无偏见的T细胞表位分析的方法，使用CD 8 +T细胞与用高复杂性、编码表位的小基因文库转导的靶细胞一起体外共培养。在细胞凋亡之前，通过荧光激活的细胞分选分离在共培养物中经受来自T细胞的细胞毒性攻击的靶细胞，并通过对编码的小基因进行测序来表征。然后，我们使用已知的鼠T细胞受体/肽-MHC对和不同的小基因编码的表位文库来验证这种高度并行化的方法。因此，我们的数据表明，这种表位分析方法允许明确和敏感的识别自然加工和MHC呈递的肽表位。
Cytotoxic CD8+T cells recognize and eliminate infected or malignant cells that present peptide epitopes derived from intracellularly processed antigens on their surface. However, comprehensive profiling of specific major histocompatibility complex (MHC)-bound peptide epitopes that are naturally processed and capable of eliciting a functional T cell response has been challenging. Here, we report a method for deep and unbiased T cell epitope profiling, using in vitro co-culture of CD8+T cells together with target cells transduced with high-complexity, epitope-encoding minigene libraries. Target cells that are subject to cytotoxic attack from T cells in co-culture are isolated prior to apoptosis by fluorescence-activated cell sorting, and characterized by sequencing the encoded minigenes. We then validate this highly parallelized method using known murine T cell receptor/peptide-MHC pairs and diverse minigene-encoded epitope libraries. Our data thus suggest that this epitope profiling method allows unambiguous and sensitive identification of naturally processed and MHC-presented peptide epitopes.