Kuca-Warnawin Ewa, Plebańczyk Magdalena, Bonek Krzysztof, Kontny Ewa
Department of Pathophysiology and Immunology, National Institute of Geriatrics, Rheumatology and Rehabilitation, Warsaw, Poland.
Clinic of Rheumatology, National Institute of Geriatrics, Rheumatology and Rehabilitation, Warsaw, Poland.
Reumatologia. 2021;59(1):12-22. doi: 10.5114/reum.2021.103940. Epub 2021 Feb 28.
T-cell-mediated adaptive immunity contributes to the development and persistence of ankylosing spondylitis (AS). Mesenchymal stromal/stem cells (MSCs) have immunomodulatory potential and are able to inhibit T-cell proliferation, but their functionality in AS patients is relatively unknown. The aim of the study was to assess the direct anti-proliferative effects of MSCs isolated from subcutaneous abdominal adipose tissue of AS patients (AS/ASCs) on allogeneic T lymphocytes, using commercially available ASC lines from healthy donors (HD/ASCs) as a control.
CD3+CD4+ T-cells were isolated from peripheral blood of healthy blood donors, activated with anti-CD3/CD28 beads, and co-cultured for 5 days with untreated and TNF+IFN-γ pre-stimulated HD/ASCs (5 cell lines) and AS/ASCs, obtained from 11 patients (6F/5M). The proliferative response of T-cells was analysed by flow cytometry, while the concentrations of kynurenines, prostaglandin E2 (PGE-2), interleukin 10 (IL-10), and interleukin 1 receptor antagonist (IL-1Ra) were measured spectrophotometrically or using a specific enzyme-linked immunosorbent assay (ELISA).
HD/ASCs and AS/ASCs similarly reduced the T-cell proliferation response, i.e. the percentage of proliferating cells, the proliferation, and replication indices, and these effects were dependent mostly on soluble factors. In the co-cultures of activated CD4+ T-cells with HD/ASCs and AS/ASCs significant increases of kynurenines, PGE-2, and IL-1Ra, but not IL-10, production were observed. The release of these factors was dependent either on cell-to-cell contact (IL-10, IL-1Ra) or soluble factors (kynurenines, PGE-2). There was a moderate to strong negative correlation between T-cell proliferative response, and the concentrations of kynurenines, PGE-2, and IL-10, but not IL-1Ra. This association was more evident in the case of TI-treated AS/ASCs than HD/ASCs.
AS/ASCs, similar to HD/ASCs, exert a direct effective anti-proliferative impact on CD4+ T cells, acting via soluble factors that are released in cell contact-dependent (IL-10) and independent (kynurenines, PGE-2) pathways. Thus, our results suggest that AS/ASCs are potentially useful for therapeutic application.
T细胞介导的适应性免疫在强直性脊柱炎（AS）的发生和持续发展中起作用。间充质基质/干细胞（MSCs）具有免疫调节潜能，能够抑制T细胞增殖，但其在AS患者中的功能相对未知。本研究旨在评估从AS患者腹部皮下脂肪组织分离的MSCs（AS/ASCs）对同种异体T淋巴细胞的直接抗增殖作用，并以健康供体的市售ASC系（HD/ASCs）作为对照。
从健康献血者外周血中分离CD3+CD4+T细胞，用抗CD3/CD28磁珠激活，然后与未经处理以及经TNF+IFN-γ预刺激的HD/ASCs（5个细胞系）和AS/ASCs（从11例患者中获得，6例女性/5例男性）共培养5天。通过流式细胞术分析T细胞的增殖反应，同时用分光光度法或特异性酶联免疫吸附测定（ELISA）测量犬尿氨酸、前列腺素E2（PGE-2）、白细胞介素10（IL-10）和白细胞介素1受体拮抗剂（IL-1Ra）的浓度。
HD/ASCs和AS/ASCs同样降低了T细胞增殖反应，即增殖细胞的百分比、增殖和复制指数，这些作用主要依赖于可溶性因子。在活化的CD4+T细胞与HD/ASCs和AS/ASCs的共培养中，观察到犬尿氨酸、PGE-2和IL-1Ra（而非IL-10）的产生显著增加。这些因子的释放要么依赖于细胞间接触（IL-10、IL-1Ra），要么依赖于可溶性因子（犬尿氨酸、PGE-2）。T细胞增殖反应与犬尿氨酸、PGE-2和IL-10（而非IL-1Ra）的浓度之间存在中度至强的负相关。在经TI处理的AS/ASCs中，这种关联比HD/ASCs中更明显。
AS/ASCs与HD/ASCs相似，通过细胞接触依赖性（IL-10）和非依赖性（犬尿氨酸、PGE-2）途径释放的可溶性因子，对CD4+T细胞产生直接有效的抗增殖作用。因此，我们的结果表明AS/ASCs在治疗应用中具有潜在用途。
