Rutgers Robert Wood Johnson Medical School, New Brunswick, NJ.
Oncology Informatics and Genomics, Philips North America, Cambridge, MA.
J Immunol. 2023 Dec 1;211(11):1630-1642. doi: 10.4049/jimmunol.2300213.
Peptide loading of MHC class II (MHCII) molecules is facilitated by HLA-DM (DM), which catalyzes CLIP release, stabilizes empty MHCII, and edits the MHCII-bound peptide repertoire. HLA-DO (DO) binds to DM and modulates its activity, resulting in an altered set of peptides presented at the cell surface. MHCII-peptide presentation in individuals with type 1 diabetes (T1D) is abnormal, leading to a breakdown in tolerance; however, no direct measurement of the MHCII pathway activity in T1D patients has been performed. In this study, we measured MHCII Ag-processing pathway activity in humans by determining MHCII, MHCII-CLIP, DM, and DO levels by flow cytometry for peripheral blood B cells, dendritic cells, and monocytes from 99 T1D patients and 97 controls. Results showed that MHCII levels were similar for all three APC subsets. In contrast, MHCII-CLIP levels, independent of sex, age at blood draw, disease duration, and diagnosis age, were significantly increased for all three APCs, with B cells showing the largest increase (3.4-fold). DM and DO levels, which usually directly correlate with MHCII-CLIP levels, were unexpectedly identical in T1D patients and controls. Gene expression profiling on PBMC RNA showed that DMB mRNA was significantly elevated in T1D patients with residual C-peptide. This resulted in higher levels of DM protein in B cells and dendritic cells. DO levels were also increased, suggesting that the MHCII pathway maybe differentially regulated in individuals with residual C-peptide. Collectively, these studies show a dysregulation of the MHCII Ag-processing pathway in patients with T1D.
MHC II 类分子(MHCII)的肽负载由 HLA-DM(DM)促进,DM 催化 CLIP 释放、稳定空 MHCII,并编辑 MHCII 结合的肽库。HLA-DO(DO)与 DM 结合并调节其活性,导致细胞表面呈现的肽集合发生改变。1 型糖尿病(T1D)患者的 MHCII-肽呈递异常,导致耐受失败;然而,尚未对 T1D 患者的 MHCII 途径活性进行直接测量。在这项研究中,我们通过流式细胞术测定外周血 B 细胞、树突状细胞和单核细胞中的 MHCII、MHCII-CLIP、DM 和 DO 水平,来测量人类 MHCII 抗原加工途径的活性,共纳入了 99 名 T1D 患者和 97 名对照者。结果表明,所有三个 APC 亚群的 MHCII 水平相似。相比之下,MHCII-CLIP 水平不受性别、采血时的年龄、疾病持续时间和诊断年龄的影响,在所有三种 APC 中均显著增加,其中 B 细胞增加幅度最大(3.4 倍)。DM 和 DO 水平通常与 MHCII-CLIP 水平直接相关,但出人意料的是,T1D 患者和对照组之间的 DM 和 DO 水平相同。PBMC RNA 的基因表达谱分析显示,具有残余 C 肽的 T1D 患者的 DMB mRNA 显著升高。这导致 B 细胞和树突状细胞中的 DM 蛋白水平升高。DO 水平也升高,表明具有残余 C 肽的个体中 MHCII 途径可能受到不同的调节。总之,这些研究表明 T1D 患者的 MHCII 抗原加工途径失调。