Massachusetts General Hospital and, Harvard Medical School, Boston, Massachusetts.
Boston University School of Medicine, Boston, Massachusetts.
Arthritis Rheumatol. 2017 May;69(5):964-975. doi: 10.1002/art.40003. Epub 2017 Apr 7.
Prevotella copri, an intestinal microbe, may overexpand in stool samples from patients with new-onset rheumatoid arthritis (RA), but it is not yet clear whether the organism has immune relevance in RA pathogenesis.
HLA-DR-presented peptides (T cell epitopes) from P copri were sought directly in the patients' synovial tissue or peripheral blood mononuclear cell (PBMC) samples using tandem mass spectrometry. The antigenicity of peptides or their source proteins was examined in samples from the RA patients or comparison groups. T cell reactivity was determined by enzyme-linked immunospot assay; antibody responses were measured by enzyme-linked immunosorbent assay, and cytokine/chemokine determinations were made by bead-based assays. Serum and synovial fluid samples were examined for 16S ribosomal DNA for P copri using nested polymerase chain reaction analysis.
In PBMCs, we identified an HLA-DR-presented peptide from a 27-kd protein of P copri (Pc-p27), which stimulated Th1 responses in 42% of patients with new-onset RA. In both new-onset RA patients and chronic RA patients, 1 subgroup had IgA antibody responses to either Pc-p27 or the whole organism, which correlated with Th17 cytokine responses and frequent anti-citrullinated protein antibodies (ACPAs). The other subgroup had IgG P copri antibodies, which were associated with Prevotella DNA in synovial fluid, P copri-specific Th1 responses, and less frequent ACPAs. In contrast, P copri antibody responses were rarely found in patients with other rheumatic diseases or in healthy controls.
Subgroups of RA patients have differential IgG or IgA immune reactivity with P copri, which appears to be specific for this disease. These observations provide evidence that P copri is immune-relevant in RA pathogenesis.
肠道微生物Prevotella copri 可能在新发类风湿关节炎(RA)患者的粪便样本中过度扩张,但尚不清楚该生物在 RA 发病机制中是否具有免疫相关性。
使用串联质谱法直接在患者的滑膜组织或外周血单核细胞(PBMC)样本中寻找 P copri 的 HLA-DR 呈递肽(T 细胞表位)。在 RA 患者或对照组的样本中检查肽或其源蛋白的抗原性。通过酶联免疫斑点测定法测定 T 细胞反应性;通过酶联免疫吸附试验测量抗体反应,并通过基于珠的测定法测定细胞因子/趋化因子。使用巢式聚合酶链反应分析检查血清和滑液样本中是否存在 P copri 的 16S 核糖体 DNA。
在 PBMC 中,我们从 P copri 的 27kD 蛋白中鉴定出一个 HLA-DR 呈递肽(Pc-p27),该肽刺激 42%的新发 RA 患者产生 Th1 反应。在新发 RA 患者和慢性 RA 患者中,1 个亚组对 Pc-p27 或整个生物体具有 IgA 抗体反应,这与 Th17 细胞因子反应和频繁的抗瓜氨酸蛋白抗体(ACPA)相关。另一个亚组具有 IgG P copri 抗体,与滑液中的 Prevotella DNA、P copri 特异性 Th1 反应和较少的 ACPA 相关。相比之下,在其他风湿性疾病患者或健康对照中很少发现 P copri 抗体反应。
RA 患者亚组具有与 P copri 不同的 IgG 或 IgA 免疫反应性,这似乎是该疾病特有的。这些观察结果为 P copri 在 RA 发病机制中具有免疫相关性提供了证据。