The effects of age, origin, and biological sex on rodent mast cell (BMMC and MC/9) and basophil (RBL-2H3) phenotype and function

IF 3.7 4区 医学 Q2 CELL BIOLOGY Cellular immunology Pub Date : 2023-09-01 DOI:10.1016/j.cellimm.2023.104751
Ashley Wagner , Syed Benazir Alam , Marianna Kulka
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引用次数: 1

Abstract

Mast cells initiate allergic inflammatory immune responses and play a role in disease by releasing various inflammatory and immunomodulatory mediators. Several mast cell-lines and primary cultured cells have been used as mast cell models with inconsistent results among research groups. Bone marrow-derived mast cells (BMMC) cultured from mouse bone marrow progenitor cells are often used as a representative model of mucosal mast cell behaviour, however their reported phenotype is variable due to inconsistent culture protocols. RBL-2H3 is a rat basophilic histamine-releasing cell line that has some characteristics of both mast cells and basophils but is not a true representation of either cell type. The murine mast cell line MC/9 is an IL-3-dependent mucosal mast cell model but has limited mast cell characteristics. In this study, we have compared the response of BMMC (derived from C57BL/6 male or female mice), two sources of RBL-2H3 (purchased directly from ATCC and a lab curated culture), and MC/9 (ATCC) at several critical stages to some common stimuli (IgE/Ag, A23187) and analyzed mast cell morphology, expression level of common mast cell surface markers (CD117 and FcεRI), protease expression, and function (growth kinetics, viability, ROS production, degranulation, cytokine release and FcεRI signaling). The objective of this study was to provide insight into the effects of culture conditions, biological sex, and age of the cells on variability among reported phenotypes and, to determine optimal conditions for activation of these cells. Our data show that factors that are often overlooked such as source, age and biological sex of mast cells play an integral role in phenotypic outcomes and may account for the reported variability in their function.

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年龄、来源和生物性别对鼠类肥大细胞(BMMC和MC/9)和嗜碱性粒细胞(RBL-2H3)表型和功能的影响
肥大细胞启动过敏性炎症免疫反应,并通过释放各种炎症和免疫调节介质在疾病中发挥作用。一些肥大细胞系和原代培养细胞已被用作肥大细胞模型,研究小组之间的结果不一致。从小鼠骨髓祖细胞培养的骨髓源性肥大细胞(BMMC)通常被用作粘膜肥大细胞行为的代表性模型,然而,由于培养方案不一致,其报告的表型是可变的。RBL-2H3是一种大鼠嗜碱性组胺释放细胞系,具有肥大细胞和嗜碱性细胞的一些特征,但不是这两种细胞类型的真实代表。小鼠肥大细胞系MC/9是IL-3依赖性粘膜肥大细胞模型,但具有有限的肥大细胞特性。在本研究中,我们比较了BMMC(来源于C57BL/6雄性或雌性小鼠)、RBL-2H3的两种来源(直接从ATCC和实验室策划的培养物购买)和MC/9(ATCC)在几个关键阶段对一些常见刺激(IgE/Ag,A23187)的反应,并分析了肥大细胞形态、常见肥大细胞表面标记物(CD117和FcεRI)的表达水平、蛋白酶表达,和功能(生长动力学、活力、ROS产生、脱颗粒、细胞因子释放和FcεRI信号传导)。本研究的目的是深入了解培养条件、生物性别和细胞年龄对所报道表型变异性的影响,并确定激活这些细胞的最佳条件。我们的数据表明,经常被忽视的因素,如肥大细胞的来源、年龄和生物学性别,在表型结果中发挥着不可或缺的作用,并可能解释了其功能的变异性。
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来源期刊
Cellular immunology
Cellular immunology 生物-免疫学
CiteScore
8.20
自引率
2.30%
发文量
102
审稿时长
30 days
期刊介绍: Cellular Immunology publishes original investigations concerned with the immunological activities of cells in experimental or clinical situations. The scope of the journal encompasses the broad area of in vitro and in vivo studies of cellular immune responses. Purely clinical descriptive studies are not considered. Research Areas include: • Antigen receptor sites • Autoimmunity • Delayed-type hypersensitivity or cellular immunity • Immunologic deficiency states and their reconstitution • Immunologic surveillance and tumor immunity • Immunomodulation • Immunotherapy • Lymphokines and cytokines • Nonantibody immunity • Parasite immunology • Resistance to intracellular microbial and viral infection • Thymus and lymphocyte immunobiology • Transplantation immunology • Tumor immunity.
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