OBJECTIVE Autoantibodies targeting histidyl-transfer RNA synthetase (HisRS; anti-Jo-1) are common in the idiopathic inflammatory myopathies (IIMs) and antisynthetase syndrome. This study was undertaken to investigate immunity against HisRS in the blood and lungs of patients with IIM/antisynthetase syndrome. METHODS Bronchoalveolar lavage (BAL) fluid, BAL fluid cells, and peripheral blood mononuclear cells (PBMCs) from patients with IIM/antisynthetase syndrome (n = 24) were stimulated with full-length HisRS protein or a HisRS-derived peptide (HisRS11-23 ). BAL fluid and PBMCs from patients with sarcoidosis (n = 7) and healthy subjects (n = 12) were included as controls. The CD4+ T cell response was determined according to levels of CD40L up-regulation and cytokine expression using flow cytometry. Anti-Jo-1 autoantibody responses in the serum and BAL fluid were assessed by enzyme-linked immunosorbent assay. Lung biopsy samples from patients with IIM/antisynthetase syndrome (n = 14) were investigated by immunohistochemistry. RESULTS In BAL fluid, CD4+ T cells from 3 of 4 patients with IIM/antisynthetase syndrome responded to stimulation with HisRS protein, as measured by the median fold change in CD40L expresssion in stimulated cells compared to unstimulated cells (median fold change 3.6, interquartile range [IQR] 2.7-14.7), and 2 of 3 patients with IIM/antisynthetase syndrome had the highest responses to HisRS11-23 (median fold change 88, IQR 27-149). In PBMCs, CD4+ T cells from 14 of 18 patients with IIM/antisynthetase syndrome responded to HisRS protein (median fold change 7.38, IQR 2.69-31.86; P < 0.001), whereas a HisRS11-23 response was present in 11 of 14 patients with IIM/antisynthetase syndrome (median fold change 3.4, IQR 1.87-10.9; P < 0.001). In the control group, there was a HisRS11-23 response in 3 of 7 patients with sarcoidosis (median fold change 2.09, IQR 1.45-3.29) and in 5 of 12 healthy controls (median fold change 2, IQR 1.89-2.42). CD4+ T cells from patients with IIM/antisynthetase syndrome displayed a pronounced Th1 phenotype in the BAL fluid when compared to the PBMCs (P < 0.001), producing high amounts of interferon-γ and interleukin-2 following stimulation. Anti-Jo-1 autoantibodies were detected in BAL fluid and germinal center (GC)-like structures were seen in the lung biopsy samples from patients with IIM/antisynthetase syndrome. CONCLUSION The results of this study demonstrate a pronounced presence of HisRS-reactive CD4+ T cells in PBMCs and BAL fluid cells from patients with IIM/antisynthetase syndrome as compared to patients with sarcoidosis and healthy controls. These findings, combined with the presence of anti-Jo-1 autoantibodies in BAL fluid and GC-like structures in the lungs, suggest that immune activation against HisRS might take place within the lungs of patients with IIM/antisynthetase syndrome.

中文翻译：

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特发性炎性肌病患者血液和肺中促炎性的组氨酸转移RNA合成酶特异性CD4 + T细胞。

目的针对组氨酸转移RNA合成酶（HisRS；抗Jo-1）的自身抗体在特发性炎症性肌病（IIM）和抗合成酶综合征中很常见。这项研究旨在调查IIM /抗合成酶综合征患者血液和肺部对HisRS的免疫力。方法用全长HisRS蛋白或HisRS衍生肽（HisRS11-23）刺激IIM /抗合成酶综合征（n = 24）患者的支气管肺泡灌洗（BAL）液，BAL液细胞和外周血单个核细胞（PBMC）。 ）。结节病患者（n = 7）和健康受试者（n = 12）的BAL液和PBMCs作为对照。使用流式细胞术根据CD40L上调水平和细胞因子表达确定CD4 + T细胞应答。通过酶联免疫吸附测定法评估血清和BAL液中的抗Jo-1自身抗体反应。通过免疫组织化学研究了来自IIM /抗合成酶综合征（n = 14）患者的肺活检样本。结果在BAL液中，IIM /抗合成酶综合征的4例患者中有3例的CD4 + T细胞对HisRS蛋白的刺激产生了反应，用刺激细胞中CD40L表达的中位数倍数变化与未刺激细胞相比（中位数倍数3.6，四分位间距[IQR] 2.7-14.7），并且3名IIM /抗合成酶综合征患者中有2名对HisRS11-23的反应最高（中位数倍数变化88，IQR 27-149）。在PBMC中，来自18位IIM /抗合成酶综合征患者中的14位的CD4 + T细胞对HisRS蛋白有反应（中位数倍数变化7.38，IQR 2.69-31.86； P <0.001），而14位IIM /抗合成酶综合征患者中有11位存在HisRS11-23反应（中位倍数变化3.4，IQR 1.87-10.9； P <0.001）。对照组中，结节病患者7例中的3例（中位数倍数变化2.09，IQR 1.45-3.29），健康对照组12例中的5例（中位数倍数变化2，IQR 1.89-2.42）有HisRS11-23反应。与PBMC相比，IIM /抗合成酶综合症患者的CD4 + T细胞在BAL液中表现出明显的Th1表型（P <0.001），刺激后产生大量的干扰素-γ和白介素-2。在BAL液中检测到抗Jo-1自身抗体，在IIM /抗合成酶综合征患者的肺活检样本中发现了生发中心（GC）样结构。结论该研究结果表明，与结节病患者和健康对照组相比，IIM /抗合成酶综合征患者的PBMC和BAL液细胞中明显存在HisRS反应性CD4 + T细胞。这些发现，再加上肺中BAL液和GC样结构中存在抗Jo-1自身抗体，表明针对HisM的免疫激活可能发生在IIM /抗合成酶综合征患者的肺部。