Martin Carla, Espaillat Mel Pilar, Santiago-Schwarz Frances
Biology Department, Farmingdale State College, State University of New York, Farmingdale, NY, 11735, USA.
Molecular Genetics and Microbiology and Department of Medicine, State University of New York at Stony Brook, Stony Brook, NY, 11794, USA.
Immunol Res. 2015 Dec;63(1-3):131-43. doi: 10.1007/s12026-015-8700-y.
An evolving premise is that cytoprotective autophagy responses are essential to monocyte-macrophage differentiation. Whether autophagy functions similarly during the monocyte-to-dendritic cell (DC) transition is unclear. IL-10, which induces apoptosis in maturing human DCs, has been shown to inhibit starvation-induced autophagy in murine macrophage cell lines. Based on the strict requirement that Bcl-2-mediated anti-apoptotic processes are implemented during the monocyte-to-DC transition, we hypothesized that cytoprotective autophagy responses also operate at the monocyte-DC interface and that IL-10 inhibits both anti-apoptotic and cytoprotective autophagy responses at this critical juncture. In support of our premise, we show that levels of anti-apoptotic Bcl-2 and autophagy-associated LC3 and Beclin-1 proteins are coincidentally upregulated during the monocyte-to-DC transition. Autophagy was substantiated by increased autophagosome visualization after bafilomycin treatment. Moreover, the autophagy inhibitor 3-MA restricted DC differentiation by prompting apoptosis. IL-10 implemented apoptosis that was coincidentally associated with reduced levels of Bcl-2 and widespread disruption of the autophagic flux. During peak apoptosis, IL-10 produced the death of newly committed DCs. However, cells surviving the IL-10 apoptotic schedule were highly phagocytic macrophage-like cells displaying reduced capacity to stimulate allogeneic naïve T cells in a mixed leukocyte reaction, increased levels of LC3, and mature autophagosomes. Thus, IL-10's negative control of DC-driven adaptive immunity at the monocyte-DC interface includes disruption of coordinately regulated molecular networks involved in pro-survival autophagy and anti-apoptotic responses.
一个不断发展的前提是细胞保护性自噬反应对于单核细胞向巨噬细胞的分化至关重要。自噬在单核细胞向树突状细胞(DC)转变过程中是否发挥类似作用尚不清楚。IL-10可诱导成熟人DC凋亡,已证明它能抑制鼠巨噬细胞系中饥饿诱导的自噬。基于在单核细胞向DC转变过程中严格需要Bcl-2介导的抗凋亡过程,我们推测细胞保护性自噬反应也在单核细胞与DC的界面发挥作用,并且IL-10在这个关键节点抑制抗凋亡和细胞保护性自噬反应。为支持我们的前提,我们表明在单核细胞向DC转变过程中,抗凋亡蛋白Bcl-2以及自噬相关蛋白LC3和Beclin-1的水平同时上调。在用巴弗洛霉素处理后自噬体可视化增加证实了自噬的存在。此外,自噬抑制剂3-MA通过促使细胞凋亡限制了DC的分化。IL-10引发的凋亡与Bcl-2水平降低以及自噬流的广泛破坏同时发生。在凋亡高峰期,IL-10导致新分化的DC死亡。然而,在IL-10诱导的凋亡过程中存活下来的细胞是高度吞噬性的巨噬细胞样细胞,它们在混合淋巴细胞反应中刺激同种异体幼稚T细胞的能力降低,LC3水平升高,并且有成熟的自噬体。因此,IL-10在单核细胞与DC界面处对DC驱动的适应性免疫的负调控包括破坏参与促存活自噬和抗凋亡反应的协调调节分子网络。