Proteomic characterization of phagocytic primary human monocyte-derived macrophages.
Proteomic characterization of phagocytic primary human monocyte-derived macrophages.
复制标题
DOI：
10.1039/d2cb00076h
复制
发表时间：
2022-06-08
影响因子：
4.1
通讯作者：
Zaro, Balyn W.
中科院分区：
文献类型：
--
作者：
Volk, Regan F.;Montano, Jose L.;Warrington, Sara E.;Hofmann, Katherine L.;Zaro, Balyn W.
巨噬细胞在先天免疫系统中发挥着至关重要的作用，识别并摧毁不需要的细胞。然而，由于技术限制，很难全面了解巨噬细胞蛋白质丰度。此外，目前还不清楚蛋白质组组成的变化如何与吞噬细胞活性相关。在这项研究中，我们开发了衍生人类巨噬细胞并准备进行质谱分析的方法，以便更深入地了解巨噬细胞刺激的蛋白质组学后果。干扰素 γ (IF-g) 是一种免疫刺激细胞因子，用于诱导巨噬细胞活化，使癌细胞的吞噬能力增加 2 倍。这些条件用于在静息巨噬细胞和吞噬活性增加的受刺激巨噬细胞之间进行鸟枪法蛋白质组学比较。我们的分析表明，巨噬细胞的蛋白质生产偏向于与吞噬作用和响应刺激的抗原加工相关的生物过程。我们通过基于抗体的蛋白质印迹实验证实了我们的发现，验证了先前报道的和新颖的感兴趣的蛋白质。除了整个蛋白质的变化之外，我们还通过用蛋氨酸替代探针高炔丙基甘氨酸 (HPG) 处理细胞来评估活性蛋白质合成。我们发现吞噬作用诱导刺激期间 HPG 掺入率增加，表明刺激改变了蛋白质合成率。我们的研究结果共同提供了迄今为止对人类原代巨噬细胞最全面的蛋白质组学见解。我们预计这些数据可以用作生成有关先天免疫功能的新假设的起点。我们报告了在原代人单核细胞衍生的巨噬细胞中可检测和合成的蛋白质的一流数据集。与稳态巨噬细胞相比，用干扰素-γ刺激的巨噬细胞具有更强的吞噬能力并合成更多的蛋白质。
Macrophages play a vital role in the innate immune system, identifying and destroying unwanted cells. However, it has been difficult to attain a comprehensive understanding of macrophage protein abundance due to technical limitations. In addition, it remains unclear how changes in proteome composition are linked to phagocytic activity. In this study we developed methods to derive human macrophages and prepare them for mass spectrometry analysis in order to more-deeply understand the proteomic consequences of macrophage stimulation. Interferon gamma (IF-g), an immune stimulating cytokine, was used to induce macrophage activation, increasing phagocytosis of cancer cells by 2-fold. These conditions were used to perform comparative shotgun proteomics between resting macrophages and stimulated macrophages with increased phagocytic activity. Our analysis revealed that macrophages bias their protein production toward biological processes associated with phagocytosis and antigen processing in response to stimulation. We confirmed our findings by antibody-based western blotting experiments, validating both previously reported and novel proteins of interest. In addition to whole protein changes, we evaluated active protein synthesis by treating cells with the methionine surrogate probe homopropargylglycine (HPG). We saw increased rates of HPG incorporation during phagocytosis-inducing stimulation, suggesting protein synthesis rates are altered by stimulation. Together our findings provide the most comprehensive proteomic insight to date into primary human macrophages. We anticipate that this data can be used as a launchpoint to generate new hypotheses about innate immune function. We report a first-in-class dataset of proteins detectable and synthesized in primary human monocyte-derived macrophages. Macrophages stimulated with interferon-gamma are more phagocytic and synthesize more proteins than steady-state macrophages.
登录
查看更多内容
影响因子：
1.2
作者：
Kumar A;Jamwal S;Midha MK;Hamza B;Aggarwal S;Yadav AK;Rao KV
通讯作者：
Rao KV
DOI：
10.1016/j.iotech.2022.100070
发表时间：
2022-03
期刊：
Immuno-oncology technology
影响因子：
--
作者：
Maute, R;Xu, J;Weissman, I L
通讯作者：
Weissman, I L
影响因子：
64.8
作者：
Kojima Y;Volkmer JP;McKenna K;Civelek M;Lusis AJ;Miller CL;Direnzo D;Nanda V;Ye J;Connolly AJ;Schadt EE;Quertermous T;Betancur P;Maegdefessel L;Matic LP;Hedin U;Weissman IL;Leeper NJ
通讯作者：
Leeper NJ
影响因子：
3.7
作者：
Ji R;Ma L;Chen X;Sun R;Zhang L;Saiyin H;Wei W
通讯作者：
Wei W
影响因子：
30.5
作者：
Piccolo V;Curina A;Genua M;Ghisletti S;Simonatto M;Sabò A;Amati B;Ostuni R;Natoli G
通讯作者：
Natoli G