HLA DaB1*15:01-DQB1*06:02-Restricted Human CD4+T Cells Are Selectively Activated With Amoxicillin-Peptide Adducts
HLA DaB1*15:01-DQB1*06:02-Restricted Human CD4+T Cells Are Selectively Activated With Amoxicillin-Peptide Adducts
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DOI：
10.1093/toxsci/kfaa128
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发表时间：
2020-11-01
影响因子：
3.8
通讯作者：
Naisbitt, Dean J.
中科院分区：
文献类型：
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作者：
Tailor, Arun;Meng, Xiaoli;Naisbitt, Dean J.
阿莫西林克拉维酸是特异质药物性肝损伤 (DILI) 的最常见原因。在 DILI 患者中检测到药物特异性 CD4+ T 细胞，提示免疫病因。此外，包括人类白细胞抗原 (HLA) DRB1*15:01-DQB1*06:02 单倍型在内的遗传关联也会影响易感性。阿莫西林形成蛋白质加合物，推测该加合物通过与赖氨酸残基结合来激活 T 细胞。然而，尚未描述此类加合物的作用。本研究旨在 (1) 研究阿莫西林修饰的 HLA-DRB1*15:01-DQB1*06:02 结合肽是否选择性激活 DILI 患者 T 细胞，以及 (2) 确定 T 细胞对抗原结构反应的性质。设计了在位置 (KP) 2-6 携带用于阿莫西林结合的赖氨酸残基的肽和用于 HLA-DRB1*15:01-DQB1*06:02 单倍型的锚。在与患者外周血单核细胞一起培养之前，通过质谱分析对阿莫西林修饰的肽进行表征。然后测试 T 细胞克隆对阿莫西林、未修饰和阿莫西林修饰肽以及结构变体的特异性。阿莫西林修饰的 KP-2 和 KP-3 肽特异性 CD4+ 克隆以剂量依赖性方式增殖并分泌干扰素 γ (IFN-gamma)、白细胞介素和/或 IL-17/IL-22，并且与阿莫西林、未修饰的肽或位置衍生物没有交叉反应性。 T 细胞反应受到 HLA II 类限制，阿莫西林修饰肽选择性结合 HLA-DRB1*15:01 和/或 DQB1*06:02。总之，我们证明阿莫西林修饰的肽与风险单倍型的两种成分结合，刺激 DILI 患者 T 细胞，并描述了 HLA 结合肽序列中亲核赖氨酸残基位置的重要性。
Amoxicillin-clavulanate is the most common cause of idiosyncratic drug-induced liver injury (DILI). Drug-specific CD4+ T cells have been detected in patients with DILI, suggestive of an immune etiology. Furthermore, genetic associations including the human leucocyte antigen (HLA) DRB1*15:01-DQB1*06:02 haplotype influence susceptibility. Amoxicillin forms protein adducts that are postulated to activate T cells, by conjugating with lysine residues. However, a role for such adducts has not been described. This study aimed to (1) investigate whether amoxicillin-modified HLA-DRB1*15:01-DQB1*06:02 binding peptides selectively activate DILI patient T cells and (2) define the nature of the T-cell response with respective to antigen structure. Peptides carrying lysine residues for amoxicillin binding in positions (KP) 2-6 and anchors for the HLA-DRB1*15:01-DQB1*06:02 haplotype were designed. The amoxicillin-modified peptides were characterized by mass spectrometry prior to culturing with patient peripheral blood mononuclear cell. T-cell clones were then tested for specificity with amoxicillin, unmodified- and amoxicillin-modified peptides, and structural variants. Amoxicillin-modified KP-2 and KP-3 peptide-specific CD4+ clones proliferated and secreted interferon gamma (IFN-gamma), interleukin perforM and/or IL-17/IL-22 in a dose-dependent manner and displayed no cross-reactivity with amoxicillin, unmodified peptide or with positional derivatives. The T cells response was HLA class II restricted and the amoxicillin-modified peptides bound selectively to HLA-DRB1*15:01 and/or DQB1*06:02. To conclude, we show that amoxicillin-modified peptides bind to both components of the risk haplotype to stimulate DILI patient T cells and describe the importance of the position of nucleophilic lysine residue in the HLA binding peptide sequence.