Immunology and Rheumatology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Vasco de Quiroga 15 Tlalpan, Mexico City, D.F. 14000, Mexico.
J Rheumatol. 2012 Apr;39(4):830-5. doi: 10.3899/jrheum.110862. Epub 2012 Feb 15.
Previous reports have shown an increase in peripheral blood mononuclear cells' (PBMC) Th17 cell subpopulation and tumor necrosis factor-α (TNF-α) secretion after in vitro stimulation with anti-CD3/CD28 or phorbol myristate acetate/ionomycin in ankylosing spondylitis (AS). The aim of our study was to determine whether there is a Th17 polarization not subjected to in vitro stimulation in patients with AS.
Nonstimulated PBMC were analyzed from 46 patients with AS, including 7 (15.2%) receiving tumor necrosis factor-α (TNF-α) inhibitors, 20 patients with rheumatoid arthritis, and 25 healthy controls. The surface phenotype of freshly isolated PBMC was determined by flow cytometry. Th1, Th2, Th17, and Treg subsets were defined as CD3+CD4+IFN-γ+, CD3+CD4+IL-4+, CD3+CD4+IL-17A+, and CD3+CD4+FoxP3+, respectively. Serum cytokines and interleukin 8 (IL-8) levels were quantified by Luminex technology.
The percentages of Th17 and Th1 cells in AS were higher than in healthy controls (7.4% ± 1.8% vs 0.7% ± 0.2% and 4.0% ± 1.3% vs 1.1% ± 0.3%, respectively; p < 0.0001). Th17 and Th1 cell subsets in patients taking TNF-α inhibitors were lower than in those naive to such therapeutics and similar to healthy controls. Serum levels of IL-6, IL-17A, TNF-α, and IL-8 were significantly higher in patients with AS compared to controls.
The percentages of Th17 and Th1 cells in PBMC without in vitro stimulation, as well as cytokine and IL-8 levels, were significantly increased in patients with AS compared with healthy controls. These T cell subsets and cytokine profiles of patients with AS taking TNF-α inhibitors were similar to those of healthy controls.
先前的报告表明，在体外用抗 CD3/CD28 或佛波醇肉豆蔻酸乙酯/离子霉素刺激后，强直性脊柱炎（AS）患者的外周血单个核细胞（PBMC）Th17 细胞亚群和肿瘤坏死因子-α（TNF-α）分泌增加。我们的研究目的是确定 AS 患者是否存在未经体外刺激的 Th17 极化。
分析了 46 例 AS 患者的非刺激 PBMC，其中包括 7 例（15.2%）接受肿瘤坏死因子-α（TNF-α）抑制剂治疗、20 例类风湿关节炎患者和 25 名健康对照者。通过流式细胞术测定新鲜分离的 PBMC 的表面表型。Th1、Th2、Th17 和 Treg 亚群分别定义为 CD3+CD4+IFN-γ+、CD3+CD4+IL-4+、CD3+CD4+IL-17A+和 CD3+CD4+FoxP3+。通过 Luminex 技术定量检测血清细胞因子和白细胞介素 8（IL-8）水平。
AS 患者的 Th17 和 Th1 细胞百分比高于健康对照组（7.4%±1.8%比 0.7%±0.2%和 4.0%±1.3%比 1.1%±0.3%；p<0.0001）。接受 TNF-α 抑制剂治疗的患者 Th17 和 Th1 细胞亚群低于未接受此类治疗的患者，与健康对照组相似。与对照组相比，AS 患者的血清 IL-6、IL-17A、TNF-α 和 IL-8 水平显著升高。
与健康对照组相比，AS 患者未经体外刺激的 PBMC 中 Th17 和 Th1 细胞的百分比以及细胞因子和 IL-8 水平显著增加。接受 TNF-α 抑制剂治疗的 AS 患者的这些 T 细胞亚群和细胞因子谱与健康对照组相似。
