Division of Rheumatology, Inflammation, and Immunity, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02210.
Infection and Immunity Program and Department of Biochemistry and Molecular Biology, Biomedicine Discovery Institute, Monash University, Clayton, VIC 3800, Australia.
Proc Natl Acad Sci U S A. 2024 Aug 20;121(34):e2321686121. doi: 10.1073/pnas.2321686121. Epub 2024 Aug 14.
To broadly measure the spectrum of cellular self-antigens for natural killer T cells (NKT), we developed a sensitive lipidomics system to analyze lipids trapped between CD1d and NKT T cell receptors (TCRs). We captured diverse antigen complexes formed in cells from natural endogenous lipids, with or without inducing endoplasmic reticulum (ER) stress. After separating protein complexes with no, low, or high CD1d-TCR interaction, we eluted lipids to establish the spectrum of self-lipids that facilitate this interaction. Although this unbiased approach identified fifteen molecules, they clustered into only two related groups: previously known phospholipid antigens and unexpected neutral lipid antigens. Mass spectrometry studies identified the neutral lipids as ceramides, deoxyceramides, and diacylglycerols, which can be considered headless lipids because they lack polar headgroups that usually form the TCR epitope. The crystal structure of the TCR-ceramide-CD1d complex showed how the missing headgroup allowed the TCR to predominantly contact CD1d, supporting a model of CD1d autoreactivity. Ceramide and related headless antigens mediated physiological TCR binding affinity, weak NKT cell responses, and tetramer binding to polyclonal human and mouse NKT cells. Ceramide and sphingomyelin are oppositely regulated components of the "sphingomyelin cycle" that are altered during apoptosis, transformation, and ER stress. Thus, the unique molecular link of ceramide to NKT cell response, along with the recent identification of sphingomyelin blockers of NKT cell activation, provide two mutually reinforcing links for NKT cell response to sterile cellular stress conditions.
为了广泛测量自然杀伤 T 细胞(NKT)的细胞自身抗原谱,我们开发了一种灵敏的脂质组学系统来分析 CD1d 和 NKT T 细胞受体(TCR)之间捕获的脂质。我们从天然内源性脂质中捕获了在细胞中形成的各种抗原复合物,无论是否诱导内质网(ER)应激。在分离没有、低或高 CD1d-TCR 相互作用的蛋白复合物后,我们洗脱脂质以建立促进这种相互作用的自身脂质谱。尽管这种无偏方法鉴定了十五种分子,但它们仅聚集为两个相关的群体:先前已知的磷脂抗原和意想不到的中性脂质抗原。质谱研究将中性脂质鉴定为神经酰胺、脱氧神经酰胺和二酰基甘油,它们可以被认为是无头脂质,因为它们缺乏通常形成 TCR 表位的极性头基团。TCR-神经酰胺-CD1d 复合物的晶体结构显示了缺失的头基团如何允许 TCR 主要与 CD1d 接触,支持 CD1d 自身反应的模型。神经酰胺和相关的无头抗原介导生理 TCR 结合亲和力、弱 NKT 细胞反应以及四聚体与多克隆人和鼠 NKT 细胞的结合。神经酰胺和鞘磷脂是“鞘磷脂循环”的相反调节成分,在细胞凋亡、转化和 ER 应激期间发生改变。因此,神经酰胺与 NKT 细胞反应的独特分子联系,以及最近发现的鞘磷脂抑制剂阻断 NKT 细胞激活,为 NKT 细胞对无菌细胞应激条件的反应提供了两个相互增强的联系。