新兴证据表明成纤维细胞在健康和疾病状态下通过产生各种蛋白质在免疫调节中扮演关键角色。在本研究中，我们首次使用Nano-LC-ESI串联质谱比较了无血清培养的人体皮肤成纤维细胞和外周血单个核细胞（PBMC）的蛋白质组。这项分析有助于更好地理解慢性炎症和癌症的潜在分子机制，这些疾病本质上伴随着生长因子缺乏。无血清培养的成纤维细胞和PBMC的蛋白质组分别由307种和294种蛋白质组成，这些蛋白质参与淋巴细胞迁移、补体激活、炎症、急性期反应和免疫调节。无血清培养的成纤维细胞主要产生胶原/胶原酶等胞外基质相关蛋白质，而PBMC产生与淋巴细胞迁移有关的黏着斑相关蛋白质，如beta-帕尔文和绑定蛋白。PBMC产生了更多样的炎症分子，如热休克蛋白，而成纤维细胞产生人类白细胞抗原G和E，这些蛋白质被认为是主要的免疫调节分子。这两种蛋白质组中共有54种蛋白质，包括血清白蛋白、淀粉样蛋白β、热休克配伍蛋白和补体C3。GeneMANIA生物信息学工具预测PBMC有418个功能，包括反应性氧化物代谢过程，而无血清培养的成纤维细胞有241个功能，如抗原处理和呈递，包括非经典MHC-Ib途径以及免疫应答的负调节。蛋白质-蛋白质相互作用网络分析表明无血清培养的成纤维细胞来源的人类白细胞抗原G和E具有免疫抑制功能。此外，在异基因移植的体外模型中，无血清培养的成纤维细胞的免疫抑制活性得到了实验证明。在血清饥饿诱导的代谢应激下，PBMC和成纤维细胞都会产生热休克蛋白和淀粉样蛋白β等分子，这些分子在类风湿性关节炎、1型糖尿病、系统性红斑狼疮、衰老和癌症等自身炎症性疾病中可能发挥致病作用。然而，无血清培养的成纤维细胞在异基因移植的体外模型中显示出免疫抑制活性，表明它们通过下调免疫系统可能有改变这些不良反应的潜力。© 2023 The Authors. Published by Elsevier Ltd.

Emerging evidence indicates that fibroblasts play pivotal roles in immunoregulation by producing various proteins under health and disease states. In the present study, for the first time, we compared the proteomes of serum-starved human skin fibroblasts and peripheral blood mononuclear cells (PBMCs) using Nano-LC-ESI-tandem mass spectrometry. This analysis contributes to a better understanding of the underlying molecular mechanisms of chronic inflammation and cancer, which are intrinsically accompanied by growth factor deficiency.The proteomes of starved fibroblasts and PBMCs consisted of 307 and 294 proteins, respectively, which are involved in lymphocyte migration, complement activation, inflammation, acute phase response, and immune regulation. Starved fibroblasts predominantly produced extracellular matrix-related proteins such as collagen/collagenase, while PBMCs produced focal adhesion-related proteins like beta-parvin and vinculin which are involved in lymphocyte migration. PBMCs produced a more diverse set of inflammatory molecules like heat shock proteins, while fibroblasts produced human leukocytes antigen-G and -E that are known as main immunomodulatory molecules. Fifty-four proteins were commonly found in both proteomes, including serum albumin, amyloid-beta, heat shock cognate 71 kDa, and complement C3. GeneMANIA bioinformatic tool predicted 418 functions for PBMCs, including reactive oxygen species metabolic processes and 241 functions for starved fibroblasts such as antigen processing and presentation including non-classical MHC -Ib pathway, and negative regulation of the immune response. Protein-protein interactions network analysis indicated the immunosuppressive function for starved fibroblasts-derived human leucocytes antigen-G and -E. Moreover, in an in vitro model of allogeneic transplantation, the immunosuppressive activity of starved fibroblasts was experimentally documented.Under serum starvation-induced metabolic stress, both PBMCs and fibroblasts produced molecules like heat shock proteins and amyloid-beta, which can have pathogenic roles in auto-inflammatory diseases such as rheumatoid arthritis, type 1 diabetes mellitus, systemic lupus erythematosus, aging, and cancer. However, starved fibroblasts showed immunosuppressive activity in an in vitro model of allogeneic transplantation, suggesting their potential to modify such adverse reactions by down-regulating the immune system.© 2023 The Authors. Published by Elsevier Ltd.