Mapping the T cell epitopes of the M-type transmembrane phospholipase A2 receptor in primary membranous nephropathy.
Mapping the T cell epitopes of the M-type transmembrane phospholipase A2 receptor in primary membranous nephropathy.
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DOI：
10.1016/j.kint.2022.11.021
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发表时间：
2022-12
影响因子：
19.6
通讯作者：
Xiao-dan Zhang;Cai-xia Lin;Z. Cui;Qiu-Hua Gu;Bi Yan;Lei Liu;W. Song;Yi Shi;H. Debiec
中科院分区：
文献类型：
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作者：
Xiao-dan Zhang;Cai-xia Lin;Z. Cui;Qiu-Hua Gu;Bi Yan;Lei Liu;W. Song;Yi Shi;H. Debiec
M型磷脂酶A2受体（PLA 2 R）是原发性膜性肾病（MN）的主要自身抗原。尽管有许多关于抗体识别的B细胞表位的研究，但对T细胞表位知之甚少。在此，我们合成了123个线性肽，每个由15-22个氨基酸组成，其中8-12个氨基酸重叠，跨越PLA 2 R的10个结构域。然后通过流式细胞术评估它们与风险性（DRB 1 β 1501，DRB 1 β 0301）和保护性（DRB 1 β 0901，DRB 1 β 0701）HLA分子的结合能力。在肽刺激后分析来自具有抗PLA 2 R阳性MN的患者的CD 4 + T细胞的增殖。通过流式细胞仪微珠阵列测量由活化的外周血单个核细胞产生的细胞因子。我们鉴定了17个PLA 2 R肽，其以高容量结合DRB 1 β 1501和DRB 1 β 0301分子。这些肽中的一些显示出与杂合DRB 1 β 1501/0901和DRB 1 β 0301/0701的结合降低。17种肽中的10种（CysR 1、CysR 10、CysR 12、FnII-3、CTLD 3 -9、CTLD 3 -10、CTLD 3 -11、CTLD 5 -2-1、CTLD 7 -1和CTLD 7 -2）诱导来自MN患者的CD 4 + T细胞比来自健康个体的细胞显著增殖。在被这些肽激活后，MN患者的外周血单核细胞产生更高水平的促炎细胞因子，主要是IL-6、TNF-α、IL-10、IL-9和IL-17。因此，我们绘制并鉴定了PLA 2 R的CysR、FnII、CTLD 3、CTLD 5和CTLD 7结构域中的十种肽作为MN的潜在T细胞表位。这些发现是开发肽特异性免疫疗法的第一步。
The M-type phospholipase A2 receptor (PLA2R) is the major autoantigen of primary membranous nephropathy (MN). Despite many studies on B-cell epitopes recognized by antibodies, little is known about T-cell epitopes. Herein, we synthesized 123 linear peptides, each consisting of 15-22 amino acids with 8-12 amino acid overlaps, across ten domains of PLA2R. Their binding capacity to risk (DRB1∗1501, DRB1∗0301) and protective (DRB1∗0901, DRB1∗0701) HLA molecules was then assessed by flow cytometry. Proliferation of CD4+ T cells from patients with anti-PLA2R positive MN was analyzed after peptide stimulation. Cytokines produced by activated peripheral blood mononuclear cells were measured by cytometric bead arrays. We identified 17 PLA2R peptides that bound to both DRB1∗1501 and DRB1∗0301 molecules with high capacity. Some of these peptides showed decreased binding to heterozygous DRB1∗1501/0901 and DRB1∗0301/0701. Ten of the 17 peptides (CysR1, CysR10, CysR12, FnII-3, CTLD3-9, CTLD3-10, CTLD3-11, CTLD5-2-1, CTLD7-1 and CTLD7-2) induced significant proliferation of CD4+ T cells from patients with MN than cells from healthy individuals. Upon activation by these peptides, peripheral blood mononuclear cells from patients with MN produced higher levels of pro-inflammatory cytokines, predominantly IL-6, TNF-α, IL-10, IL-9 and IL-17. Thus, we mapped and identified ten peptides in the CysR, FnII, CTLD3, CTLD5, and CTLD7 domains of PLA2R as potential T-cell epitopes of MN. These findings are a first step towards developing peptide-specific immunotherapies.