Tissue Engineering Laboratory, BIH Center for Regenerative Therapies, Department for Rheumatology and Clinical Immunology & Berlin Institute of Health at Charité-Universitätsmedizin Berli, BCRT, Charitéplatz 1, 10117, Berlin, Germany.
Institute of Medical Immunology and Berlin Institute of Health Center for Regenerative Therapies, Institute of Medical Immunology, Charité-Universitaetsmedizin Berlin, corporate member of Freie Universitaet Berlin and Humboldt-Universitaet Zu Berlin, Augustenburger Platz 1, 13353, Berlin, Germany.
J Nanobiotechnology. 2021 Mar 25;19(1):83. doi: 10.1186/s12951-021-00830-7.
Chemokine therapy with C-C motif chemokine ligand 25 (CCL25) is currently under investigation as a promising approach to treat articular cartilage degeneration. We developed a delayed release mechanism based on Poly (lactic-co-glycolic acid) (PLGA) microparticle encapsulation for intraarticular injections to ensure prolonged release of therapeutic dosages. However, CCL25 plays an important role in immune cell regulation and inflammatory processes like T-cell homing and chronic tissue inflammation. Therefore, the potential of CCL25 to activate immune cells must be assessed more thoroughly before further translation into clinical practice. The aim of this study was to evaluate the reaction of different immune cell subsets upon stimulation with different dosages of CCL25 in comparison to CCL25 released from PLGA particles.
Immune cell subsets were treated for up to 5 days with CCL25 and subsequently analyzed regarding their cytokine secretion, surface marker expression, polarization, and migratory behavior. The CCL25 receptor C-C chemokine receptor type 9 (CCR9) was expressed to a different extent on all immune cell subsets. Direct stimulation of peripheral blood mononuclear cells (PBMCs) with high dosages of CCL25 resulted in strong increases in the secretion of monocyte chemoattractant protein-1 (MCP-1), interleukin-8 (IL-8), interleukin-1β (IL-1β), tumor-necrosis-factor-α (TNF-α) and interferon-γ (IFN-γ), upregulation of human leukocyte antigen-DR (HLA-DR) on monocytes and CD4 T-cells, as well as immune cell migration along a CCL25 gradient. Immune cell stimulation with the supernatants from CCL25 loaded PLGA microparticles caused moderate increases in MCP-1, IL-8, and IL-1β levels, but no changes in surface marker expression or migration. Both CCL25-loaded and unloaded PLGA microparticles induced an increase in IL-8 and MCP-1 release in PBMCs and macrophages, and a slight shift of the surface marker profile towards the direction of M2-macrophage polarization.
While supernatants of CCL25 loaded PLGA microparticles did not provoke strong inflammatory reactions, direct stimulation with CCL25 shows the critical potential to induce global inflammatory activation of human leukocytes at certain concentrations. These findings underline the importance of a safe and reliable release system in a therapeutic setup. Failure of the delivery system could result in strong local and systemic inflammatory reactions that could potentially negate the benefits of chemokine therapy.
趋化因子疗法使用 C-C 基序趋化因子配体 25（CCL25）作为治疗关节软骨退化的一种有前途的方法正在研究中。我们开发了一种基于聚（乳酸-共-乙醇酸）（PLGA）微球包封的延迟释放机制，用于关节内注射，以确保治疗剂量的长时间释放。然而，CCL25 在免疫细胞调节和炎症过程中发挥着重要作用，如 T 细胞归巢和慢性组织炎症。因此，在进一步转化为临床实践之前，必须更彻底地评估 CCL25 激活免疫细胞的潜力。本研究的目的是评估不同免疫细胞亚群在受到不同剂量 CCL25 刺激时的反应，并与从 PLGA 颗粒中释放的 CCL25 进行比较。
免疫细胞亚群用 CCL25 处理长达 5 天，然后分析其细胞因子分泌、表面标志物表达、极化和迁移行为。所有免疫细胞亚群均以不同程度表达趋化因子受体 9（CCR9）。高剂量 CCL25 直接刺激外周血单核细胞（PBMCs）会导致单核细胞趋化蛋白-1（MCP-1）、白细胞介素-8（IL-8）、白细胞介素-1β（IL-1β）、肿瘤坏死因子-α（TNF-α）和干扰素-γ（IFN-γ）的分泌显著增加，单核细胞和 CD4 T 细胞上的人类白细胞抗原-DR（HLA-DR）上调，以及沿 CCL25 梯度的免疫细胞迁移。用 CCL25 加载的 PLGA 微球上清液刺激免疫细胞会导致 MCP-1、IL-8 和 IL-1β 水平适度增加，但表面标志物表达或迁移没有变化。负载和未负载 CCL25 的 PLGA 微球均会导致 PBMC 和巨噬细胞中 IL-8 和 MCP-1 的释放增加，并使表面标志物谱向 M2 巨噬细胞极化方向略有偏移。
虽然 CCL25 加载的 PLGA 微球上清液没有引起强烈的炎症反应，但 CCL25 的直接刺激显示出在某些浓度下诱导人类白细胞整体炎症激活的关键潜力。这些发现强调了在治疗环境中安全可靠的释放系统的重要性。递送系统的失败可能导致强烈的局部和全身炎症反应，从而可能抵消趋化因子治疗的益处。
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