School of Biological Sciences, University of Northern Colorado, Greeley, Colorado, United States of America.
Olson Center for Women's Health, Department of Obstetrics and Gynecology, University of Nebraska Medical Center, Omaha, Nebraska, United States of America.
PLoS One. 2018 Nov 16;13(11):e0207704. doi: 10.1371/journal.pone.0207704. eCollection 2018.
For immune cells transforming growth factor beta-1 (TGF-β1) can enhance or repress effector functions. Here, we characterize the effects of TGF-β1 on IgE-mediated and IL-33-mediated activation of primary murine mast cells derived from hematopoietic stem cells (bone marrow derived mast cells; BMMC). We also investigated potential interactions between TGF-β1 and stem cell factor (SCF). We conclude TGF-β1 plays a selectively stimulatory role for mast cell cultures in vitro.
BMMCs from C57BL/6 mice were differentiated with IL-3 and then treated with TGF-β1. BMMCs were exposed to TGF-β1, primed with IgE, activated with antigen, and then IL-6 and IL-13 cytokine release was quantified using ELISA. Additionally, the effects of TGF-β1 on both IgE and IL-33-mediated short term activation were observed via flow cytometric analysis of both surface LAMP-1 expression and intracellular IL-6. Receptor colocalization was visualized using fluorescence confocal microscopy and individual receptor expression levels were also quantified.
Resting IL-6 production increased with TGF-β1 but significance was lost following BMMC activation via IgE receptor (FcεRI) crosslinking. This was similar to a comparison effect due to SCF treatment alone, which also enhanced resting levels of IL-6. TGF-β1 treatment enhanced release of IL-13 only with FcεRI-IgE-mediated activation. TGF-β1 suppressed mobilization of IL-6 with short-term BMMC activation when stimulated with IL-33. Lastly, colocalization patterns of the SCF receptor (CD117) and FcεRI with IgE crosslinking were unaffected by TGF-β1 treatment, but individual expression levels for FcεRI, CD117, and TGFβRII were all reduced following either IgE activation or TGF-β1 treatment; this reduction was partially recovered in BMMCs that were both activated by IgE and treated with TGF-β1.
These data reveal a novel positive effect of soluble TGF-β1 on mast cell activation in vitro, suggesting mast cells may be activated through a non-canonical pathway by TGF-β1. Understanding this interaction will provide insight into the potential role of mast cells in settings where TGF-β1 is produced in an aberrant manner, such as in and around high grade tumors.
对于免疫细胞来说，转化生长因子-β1（TGF-β1）可以增强或抑制效应功能。在这里，我们描述了 TGF-β1 对源自造血干细胞（骨髓衍生的肥大细胞；BMMC）的原代小鼠肥大细胞的 IgE 介导和 IL-33 介导的激活的影响。我们还研究了 TGF-β1 与干细胞因子（SCF）之间的潜在相互作用。我们的结论是，TGF-β1 在体外对肥大细胞培养具有选择性的刺激作用。
用 IL-3 从 C57BL/6 小鼠中分化出 BMMC，然后用 TGF-β1 处理。将 BMMC 暴露于 TGF-β1 中，用 IgE 预激活，用抗原激活，然后使用 ELISA 定量测定细胞因子 IL-6 和 IL-13 的释放。此外，通过流式细胞术分析表面 LAMP-1 表达和细胞内 IL-6，观察 TGF-β1 对 IgE 和 IL-33 介导的短期激活的影响。使用荧光共聚焦显微镜观察受体共定位，并量化单个受体的表达水平。
静息状态下的 IL-6 产生随着 TGF-β1 的增加而增加，但在 IgE 受体（FcεRI）交联后通过 BMMC 激活时失去了意义。这与单独使用 SCF 治疗的比较效果相似，后者也增强了 IL-6 的静息水平。TGF-β1 处理仅在 FcεRI-IgE 介导的激活时增强 IL-13 的释放。TGF-β1 抑制了 IL-33 刺激下短期 BMMC 激活时的 IL-6 动员。最后，用 IgE 交联处理后，SCF 受体（CD117）和 FcεRI 与 IgE 的共定位模式不受 TGF-β1 处理的影响，但在 IgE 激活或 TGF-β1 处理后，FcεRI、CD117 和 TGFβRII 的单个表达水平均降低；在 IgE 激活和 TGF-β1 处理的 BMMC 中，这种降低部分得到恢复。
这些数据揭示了可溶性 TGF-β1 对体外肥大细胞激活的新的积极影响，表明肥大细胞可能通过 TGF-β1 的非经典途径被激活。了解这种相互作用将为理解肥大细胞在 TGF-β1 以异常方式产生的情况下（例如在高级别肿瘤内及其周围）的潜在作用提供依据。
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