Thiel A, Wu P, Lauster R, Braun J, Radbruch A, Sieper J
Benjamin Franklin University Hospital, and German Rheumatology Research Center, Berlin.
Arthritis Rheum. 2000 Dec;43(12):2834-42. doi: 10.1002/1529-0131(200012)43:12<2834::AID-ANR25>3.0.CO;2-7.
In reactive arthritis (ReA) a bacteria-specific T cell response to the triggering microbe is detected in synovial fluid (SF), and an impaired Th1 cytokine response has been described. The recent identification of immunodominant bacterial proteins/peptides and new technologies make a more detailed analysis of the immune response possible. The aim of the present study was to use these new techniques to determine the antigen-specific T cell frequency and the cytokine secretion pattern on stimulation with bacteria-derived recombinant proteins in the peripheral blood (PB) and SF from patients with ReA.
In 3 patients with Chlamydia-induced ReA and 2 patients with Yersinia-induced ReA, the SF T cell response was investigated after stimulation with the Chlamydia-derived proteins major outer membrane protein (MOMP) and heat-shock protein 60 (Hsp60) and the Yersinia-derived proteins 19-kd protein and Hsp60. In 3 of these patients, the PB T cell response was investigated in parallel. T cells were stimulated in whole blood or whole SF with antigen plus anti-CD28 for 6 hours, brefeldin A was added after 2 hours, and cells were fixed and stained with antibodies against the surface markers CD4 and CD69 and against the cytokines interferon-gamma (IFNgamma), tumor necrosis factor alpha, interleukin-10 (IL-10), and IL-4. Positive cells were quantified by flow cytometry.
In the 3 patients with Chlamydia-induced ReA, the antigen-specific T cell frequency (percentage of IFNgamma CD69 double-positive CD4+ T cells) in response to MOMP (mean +/- SD 1.2 +/- 1.38%) and to Hsp60 (1.21 +/- 1.45%) in SF was about the same. In the 2 patients with Yersinia-induced ReA, the mean +/- SD frequency was 0.66 +/- 0.36% in response to the Hsp60 and 03% +/- 0.22 in response to the 19-kd protein. In the 3 patients whose PB was evaluated, the corresponding T cell response was > or =10 times lower. In 2 patients with Chlamydia-induced ReA, antigen-specific IL-10-positive CD4+ T cells were detected in 0.10-0.23% of the CD4+ T cell subpopulation.
The frequency of antigen-specific T cells to Chlamydia- and Yersinia-derived antigens in the SF of ReA patients is between 1:200 and 1:50. Both the chlamydial Hsp60 and MOMP are dominant T cell antigens in Chlamydia-induced ReA. In patients with Chlamydia-induced ReA, we detected antigen-specific IL-10 secretion, which might mediate an inhibition of effective bacterial clearance.
在反应性关节炎（ReA）中，在滑液（SF）中检测到针对引发微生物的细菌特异性T细胞反应，并且已经描述了Th1细胞因子反应受损。免疫显性细菌蛋白/肽的最新鉴定和新技术使得对免疫反应进行更详细的分析成为可能。本研究的目的是使用这些新技术来确定ReA患者外周血（PB）和SF中细菌衍生重组蛋白刺激后的抗原特异性T细胞频率和细胞因子分泌模式。
在3例衣原体诱导的ReA患者和2例耶尔森菌诱导的ReA患者中，用衣原体衍生蛋白主要外膜蛋白（MOMP）和热休克蛋白60（Hsp60）以及耶尔森菌衍生蛋白19-kd蛋白和Hsp60刺激后，研究SF T细胞反应。在其中3例患者中，并行研究PB T细胞反应。用抗原加抗CD28在全血或全SF中刺激T细胞6小时，2小时后加入布雷菲德菌素A，然后固定细胞并用抗表面标志物CD4和CD69以及抗细胞因子干扰素-γ（IFNγ）、肿瘤坏死因子α、白细胞介素-10（IL-10）和IL-4的抗体染色。通过流式细胞术对阳性细胞进行定量。
在3例衣原体诱导的ReA患者中，SF中对MOMP（平均±标准差1.2±1.38%）和Hsp60（1.21±1.45%）反应的抗原特异性T细胞频率（IFNγ CD69双阳性CD4+T细胞百分比）大致相同。在2例耶尔森菌诱导的ReA患者中，对Hsp60反应的平均±标准差频率为0.66±0.36%，对19-kd蛋白反应的频率为±0.22%。在评估PB的3例患者中，相应的T细胞反应低10倍或更多。在2例衣原体诱导的ReA患者中，在0.10 - 0.23%的CD4+T细胞亚群中检测到抗原特异性IL-10阳性CD4+T细胞。
ReA患者SF中针对衣原体和耶尔森菌衍生抗原的抗原特异性T细胞频率在1:200至1:50之间。衣原体Hsp60和MOMP都是衣原体诱导的ReA中的主要T细胞抗原。在衣原体诱导的ReA患者中，我们检测到抗原特异性IL-10分泌，这可能介导对有效细菌清除的抑制。
