Amsterdam Rheumatology & immunology Centre (ARC), Department of Clinical Immunology and Rheumatology, Academic Medical Centre, University of Amsterdam, Meibergdreef 9, Amsterdam, 1105, AZ, the Netherlands.
Department of Experimental Immunology, Academic Medical Centre, University of Amsterdam, Meibergdreef 9, Amsterdam, 1105, AZ, the Netherlands.
Arthritis Res Ther. 2018 Feb 26;20(1):35. doi: 10.1186/s13075-018-1529-8.
Systemic autoimmunity can be present years before clinical onset of rheumatoid arthritis (RA). Adaptive immunity is initiated in lymphoid tissue where lymph node stromal cells (LNSCs) regulate immune responses through their intimate connection with leucocytes. We postulate that malfunctioning of LNSCs creates a microenvironment in which normal immune responses are not properly controlled, possibly leading to autoimmune disease. In this study we established an experimental model for studying the functional capacities of human LNSCs during RA development.
Twenty-four patients with RA, 23 individuals positive for autoantibodies but without clinical disease (RA risk group) and 14 seronegative healthy control subjects underwent ultrasound-guided inguinal lymph node (LN) biopsy. Human LNSCs were isolated and expanded in vitro for functional analyses. In analogous co-cultures consisting of LNSCs and peripheral blood mononuclear cells, αCD3/αCD28-induced T-cell proliferation was measured using carboxyfluorescein diacetate succinimidyl ester dilution.
Fibroblast-like cells expanded from the LN biopsy comprised of fibroblastic reticular cells (gp38CD31) and double-negative (gp38CD31) cells. Cultured LNSCs stably expressed characteristic adhesion molecules and cytokines. Basal expression of C-X-C motif chemokine ligand 12 (CXCL12) was lower in LNSCs from RA risk individuals than in those from healthy control subjects. Key LN chemokines C-C motif chemokine ligand (CCL19), CCL21 and CXCL13 were induced in LNSCs upon stimulation with tumour necrosis factor-α and lymphotoxin αβ, but to a lesser extent in LNSCs from patients with RA. The effect of human LNSCs on T-cell proliferation was ratio-dependent and altered in RA LNSCs.
Overall, we developed an experimental model to facilitate research on the role of LNSCs during the earliest phases of RA. Using this innovative model, we show, for the first time to our knowledge, that the LN stromal environment is changed during the earliest phases of RA, probably contributing to deregulated immune responses early in disease pathogenesis.
系统性自身免疫可在类风湿关节炎 (RA) 临床发病前数年出现。适应性免疫在淋巴组织中启动，其中淋巴结基质细胞 (LNSC) 通过与白细胞的密切连接来调节免疫反应。我们假设 LNSC 功能障碍会产生一种微环境，其中正常的免疫反应无法得到适当控制，可能导致自身免疫性疾病。在这项研究中，我们建立了一个实验模型，用于研究 RA 发展过程中人类 LNSC 的功能能力。
24 例 RA 患者、23 例自身抗体阳性但无临床疾病（RA 风险组）和 14 例血清阴性健康对照者接受了腹股沟淋巴结 (LN) 超声引导活检。分离并体外扩增人 LNSC 进行功能分析。在由 LNSC 和外周血单核细胞组成的类似共培养物中，通过羧基荧光素二乙酸琥珀酰亚胺酯稀释测量 αCD3/αCD28 诱导的 T 细胞增殖。
从 LN 活检中扩增的成纤维细胞样细胞由纤维网状细胞 (gp38CD31) 和双阴性 (gp38CD31) 细胞组成。培养的 LNSC 稳定表达特征性黏附分子和细胞因子。RA 风险个体的 LNSC 中，趋化因子 C-X-C 基序配体 12 (CXCL12) 的基础表达低于健康对照者。在 TNF-α 和淋巴毒素 αβ 刺激下，关键 LN 趋化因子 C-C 基序趋化因子配体 (CCL19)、CCL21 和 CXCL13 在 LNSC 中被诱导，但在 RA 患者的 LNSC 中诱导程度较低。人 LNSC 对 T 细胞增殖的影响呈比例依赖性，且在 RA LNSC 中发生改变。
总的来说，我们开发了一种实验模型，以促进对 RA 早期阶段 LNSC 作用的研究。使用这种创新模型，我们首次表明，在 RA 的早期阶段，LN 基质环境发生了变化，这可能导致疾病发病机制早期的免疫反应失调。
