1 Queensland Children's Medical Research Institute, Royal Children's Hospital, Brisbane, The University of Queensland, Brisbane, Queensland, Australia; and.
2 Telethon Kids Institute, The University of Western Australia, Perth, Western Australia, Australia.
Am J Respir Cell Mol Biol. 2015 Nov;53(5):676-88. doi: 10.1165/rcmb.2015-0012OC.
Macrophages are dynamic cells that mature under the influence of signals from the local microenvironment into either classically (M1) or alternatively (M2) activated macrophages with specific functional and phenotypic properties. Although the phenotypic identification of M1 and M2 macrophages is well established in mice, this is less clear for human macrophages. In addition, the persistence and reversibility of polarized human phenotypes is not well established. Human peripheral blood monocytes were differentiated into uncommitted macrophages (M0) and then polarized to M1 and M2 phenotypes using LPS/IFN-γ and IL-4/IL-13, respectively. M1 and M2 were identified as CD64(+)CD80(+) and CD11b(+)CD209(+), respectively, by flow cytometry. Polarized M1 cells secreted IP-10, IFN-γ, IL-8, TNF-α, IL-1β, and RANTES, whereas M2 cells secreted IL-13, CCL17, and CCL18. Functionally, M2 cells were highly endocytic. In cytokine-deficient medium, the polarized macrophages reverted back to the M0 state within 12 days. If previously polarized macrophages were given the alternative polarizing stimulus after 6 days of resting in cytokine-deficient medium, a switch in polarization was seen (i.e., M1 macrophages switched to M2 and expressed CD11b(+)CD209(+) and vice versa). In summary, we report phenotypic identification of human M1 and M2 macrophages, their functional characteristics, and their ability to be reprogrammed given the appropriate stimuli.
巨噬细胞是一种在局部微环境信号影响下成熟的动态细胞,可以分化为经典激活型(M1)或替代激活型(M2)巨噬细胞,具有特定的功能和表型特征。虽然 M1 和 M2 巨噬细胞的表型鉴定在小鼠中已经得到很好的建立,但在人类巨噬细胞中则不太明确。此外,极化的人类表型的持久性和可逆性也尚未得到很好的建立。我们将人外周血单核细胞分化为未分化的巨噬细胞(M0),然后分别用 LPS/IFN-γ 和 IL-4/IL-13 将其极化到 M1 和 M2 表型。通过流式细胞术,M1 和 M2 分别被鉴定为 CD64(+)CD80(+)和 CD11b(+)CD209(+)。极化的 M1 细胞分泌 IP-10、IFN-γ、IL-8、TNF-α、IL-1β 和 RANTES,而 M2 细胞分泌 IL-13、CCL17 和 CCL18。功能上,M2 细胞具有高吞噬能力。在细胞因子缺乏的培养基中,极化的巨噬细胞在 12 天内会恢复到 M0 状态。如果在细胞因子缺乏的培养基中静止 6 天后,先前极化的巨噬细胞给予替代的极化刺激,则会发生极化转换(即 M1 巨噬细胞转换为 M2,表达 CD11b(+)CD209(+),反之亦然)。总之,我们报告了人类 M1 和 M2 巨噬细胞的表型鉴定、它们的功能特征以及在给予适当刺激时重新编程的能力。