Department of Clinical Immunology and Rheumatology, Academic Medical Center/University of Amsterdam, Amsterdam, The Netherlands.
PLoS One. 2012;7(4):e35994. doi: 10.1371/journal.pone.0035994. Epub 2012 Apr 26.
Costimulation of murine macrophages with immune complexes (ICs) and TLR ligands leads to alternative activation. Studies on human myeloid cells, however, indicate that ICs induce an increased pro-inflammatory cytokine production. This study aimed to clarify the effect of ICs on the pro- versus anti-inflammatory profile of human polarized macrophages.
Monocytes isolated from peripheral blood of healthy donors were polarized for four days with IFN-γ, IL-4, IL-10, GM-CSF, M-CSF, or LPS, in the presence or absence of heat aggregated gamma-globulins (HAGGs). Phenotypic polarization markers were measured by flow cytometry. Polarized macrophages were stimulated with HAGGs or immobilized IgG alone or in combination with TLR ligands. TNF, IL-6, IL-10, IL-12, and IL-23 were measured by Luminex and/or RT-qPCR.
HAGGs did not modulate the phenotypic polarization and the cytokine production of macrophages. However, HAGGs significantly altered the TLR-induced cytokine production of all polarized macrophage subsets, with the exception of MΦ(IL-4). In particular, HAGGs consistently enhanced the TLR-induced IL-10 production in both classically and alternatively polarized macrophages (M1 and M2). The effect of HAGGs on TNF and IL-6 production was less pronounced and depended on the polarization status, while IL-23p19 and IL-12p35 expression was not affected. In contrast with HAGGs, immobilized IgG induced a strong upregulation of not only IL-10, but also TNF and IL-6.
HAGGs alone do not alter the phenotype and cytokine production of in vitro polarized human macrophages. In combination with TLR-ligands, however, HAGGs but not immobilized IgG shift the cytokine production of distinct macrophage subsets toward IL-10.
用免疫复合物（ICs）和 TLR 配体刺激小鼠巨噬细胞可导致其发生替代性激活。然而，关于人类髓样细胞的研究表明，ICs 可诱导促炎细胞因子的产生增加。本研究旨在阐明 ICs 对人极化巨噬细胞促炎与抗炎表型的影响。
从健康供体的外周血中分离单核细胞，在 IFN-γ、IL-4、IL-10、GM-CSF、M-CSF 或 LPS 的存在或不存在下，用热聚合 γ-球蛋白（HAGGs）孵育 4 天进行极化。通过流式细胞术测量表型极化标志物。用 HAGGs 或单独固定 IgG 或与 TLR 配体联合刺激极化巨噬细胞。通过 Luminex 和/或 RT-qPCR 测量 TNF、IL-6、IL-10、IL-12 和 IL-23。
HAGGs 不调节巨噬细胞的表型极化和细胞因子产生。然而，HAGGs 显著改变了所有极化巨噬细胞亚群的 TLR 诱导的细胞因子产生，除了 MΦ（IL-4）。特别是，HAGGs 一致增强了经典和替代性极化巨噬细胞（M1 和 M2）中 TLR 诱导的 IL-10 产生。HAGGs 对 TNF 和 IL-6 产生的影响则不那么明显，且依赖于极化状态，而 IL-23p19 和 IL-12p35 的表达不受影响。与 HAGGs 相反，固定化 IgG 不仅强烈上调了 IL-10，还上调了 TNF 和 IL-6。
单独的 HAGGs 不会改变体外极化的人巨噬细胞的表型和细胞因子产生。然而，与 TLR 配体联合使用时，HAGGs 而非固定化 IgG 会使不同巨噬细胞亚群的细胞因子产生向 IL-10 转移。
