Duogenic StemCells Corporation, Taichung, Taiwan.
Department of Orthopedics, Show Chwan Memorial Hospital, Changhua, Taiwan.
Sci Rep. 2022 Dec 15;12(1):21663. doi: 10.1038/s41598-022-25764-4.
The human type II collagen (Col II), specifically expressed in chondrocytes, is a crucial component of the adult hyaline cartilage. We examine the potential of artificial induction of Col II in human peripheral blood mononuclear cells (PBMNCs) as a novel Col II provider. Human PBMNCs were purified and were treated with high doses of macrophage-colony stimulating factor (M-CSF), granulocyte macrophage-colony stimulating factor (GM-CSF), or granulocyte-colony stimulating factor (G-CSF) and examined the Col II expression at indicated days. Quantitative Col II expression was validated by real-time reverse transcriptase-polymerase chain reaction (RT-PCR), immunocytochemistry, and flow cytometry. We demonstrate that monocytes in PBMNCs can be artificially induced to express both Col II proteins and M2 macrophage markers by the high concentration of colony-stimulating factors, especially M-CSF and GM-CSF. The Col II proteins were detected on the cell membrane and in the cytoplasm by flow cytometry and immunocytostaining. Combination with IL-4 provided a synergistic effect with M-CSF/GM-CSF to trigger Col II expression in M2 macrophages. These CD206 and Col II double-expressing cells, named modified macrophages, share M2 macrophages' anti-inflammatory potency. We demonstrated that the modified macrophages could significantly attenuate the inflammatory progress of Complete Freund's adjuvant (CFA)-induced arthritis and collagen-induced arthritis in rodents. Here, we provide the first evidence that a modified macrophage population could ectopically express Col II and control the progress of arthritis in animals.
人Ⅱ型胶原蛋白(Col II),主要在软骨细胞中表达,是成年透明软骨的重要组成部分。我们研究了在人外周血单核细胞(PBMNCs)中人工诱导 Col II 产生的可能性,作为 Col II 的新型来源。纯化人 PBMNCs 后,用高剂量的巨噬细胞集落刺激因子(M-CSF)、粒细胞巨噬细胞集落刺激因子(GM-CSF)或粒细胞集落刺激因子(G-CSF)处理,并在指定天数检查 Col II 的表达。通过实时逆转录聚合酶链反应(RT-PCR)、免疫细胞化学和流式细胞术验证 Col II 的定量表达。我们证明 PBMNCs 中的单核细胞可以通过高浓度的集落刺激因子,特别是 M-CSF 和 GM-CSF,被人工诱导表达 Col II 蛋白和 M2 巨噬细胞标志物。Col II 蛋白通过流式细胞术和免疫细胞化学染色在细胞膜和细胞质中被检测到。与 IL-4 联合使用可与 M-CSF/GM-CSF 产生协同作用,触发 M2 巨噬细胞中 Col II 的表达。这些 CD206 和 Col II 双表达细胞,称为改良巨噬细胞,具有 M2 巨噬细胞的抗炎能力。我们证明改良巨噬细胞可以显著减轻完全弗氏佐剂(CFA)诱导的关节炎和胶原诱导的关节炎在啮齿动物中的炎症进展。在这里,我们首次提供证据表明,一种改良的巨噬细胞群体可以异位表达 Col II,并控制动物关节炎的进展。