Shen Hui, Goodall Jane C, Hill Gaston J S
University of Cambridge, Department of Medicine, and Addenbrooke's Hospital, Cambridge, UK.
Arthritis Rheum. 2009 Jun;60(6):1647-56. doi: 10.1002/art.24568.
To analyze the frequency, surface phenotype, and cytokine secretion of CD4+ T cells in peripheral blood mononuclear cells (PBMCs) from patients with ankylosing spondylitis (AS) compared with both healthy control subjects and patients with rheumatoid arthritis (RA).
Eight-color flow cytometry was used to analyze the surface phenotype and cytokine production of PBMCs from 20 patients with AS, 12 patients with RA, and 16 healthy control subjects, following stimulation ex vivo with phorbol myristate acetate and ionomycin for 5 hours. Secretion of interleukin-17 (IL-17) by PBMCs was measured by enzyme-linked immunosorbent assay, following stimulation with anti-CD3/CD28 for 4 days.
The percentages of IL-17-positive CD4+ T cells and IL-22-positive CD4+ T cells were increased in the PBMCs of both patients with AS and patients with RA compared with healthy control subjects, whereas there were no differences in the percentages of interferon-gamma (IFNgamma)-positive or IL-10-positive CD4+ T cells. Likewise, concentrations of IL-17 in supernatants from patients with AS were significantly higher compared with those from healthy control subjects. In patients with RA, the concentrations of IL-17 were increased but not significantly. There was a correlation between the percentages of IL-17-positive CD4+ T cells detected in PBMCs and the amounts of IL-17 in culture supernatants (r=0.414, P=0.0034). All IL-17-producing cells were CD4+CD45RO+; most expressed both CCR6 and CCR4, but only 50% expressed the IL-23 receptor (IL-23R). Nevertheless, there was a positive relationship between the percentage of IL-23R-positive CD4+ T cells and the frequency of IL-17-positive CD4+ T cells or IL-22-positive CD4+ T cells (r=0.57, P<0.0001 and r=0.46, P=0.001, respectively). A significant proportion of cells that produced IL-17 also produced IL-22 and IFNgamma, but none produced IL-10.
The frequencies of IL-17-positive and IL-22-positive CD4+ T cells were increased in PBMCs from patients with AS and patients with RA, resulting in secretion of higher quantities of IL-17 by PBMCs following stimulation. These data support the hypothesis that Th17 cells, particularly when present in excess of IL-10-producing cells, are involved in the pathogenesis of inflammatory arthritis.
分析强直性脊柱炎（AS）患者外周血单个核细胞（PBMC）中CD4⁺T细胞的频率、表面表型及细胞因子分泌情况，并与健康对照者和类风湿关节炎（RA）患者进行比较。
采用八色流式细胞术分析20例AS患者、12例RA患者和16例健康对照者的PBMC经佛波酯肉豆蔻酸酯乙酸盐和离子霉素体外刺激5小时后的表面表型及细胞因子产生情况。用酶联免疫吸附测定法检测PBMC经抗CD3/CD28刺激4天后白细胞介素-17（IL-17）的分泌情况。
与健康对照者相比，AS患者和RA患者PBMC中IL-17阳性CD4⁺T细胞和IL-22阳性CD4⁺T细胞的百分比均升高，而干扰素-γ（IFNγ）阳性或IL-10阳性CD4⁺T细胞的百分比无差异。同样，AS患者上清液中IL-17的浓度显著高于健康对照者。RA患者中IL-17的浓度升高但不显著。PBMC中检测到的IL-17阳性CD4⁺T细胞百分比与培养上清液中IL-17的量之间存在相关性（r=0.414，P=0.0034）。所有产生IL-17的细胞均为CD4⁺CD45RO⁺；大多数细胞同时表达CCR6和CCR4，但只有50%表达白细胞介素-23受体（IL-23R）。然而，IL-23R阳性CD4⁺T细胞百分比与IL-17阳性CD4⁺T细胞或IL-22阳性CD4⁺T细胞频率之间存在正相关（分别为r=0.57，P<0.0001和r=0.46，P=0.001）。相当一部分产生IL-17的细胞也产生IL-22和IFNγ，但均不产生IL-10。
AS患者和RA患者PBMC中IL-17阳性和IL-22阳性CD4⁺T细胞的频率升高，导致PBMC刺激后分泌更多的IL-17。这些数据支持Th17细胞，特别是当其数量超过产生IL-10的细胞时，参与炎症性关节炎发病机制的假说。
