Department of Microbiology and Immunology, University of North Carolina at Chapel Hill (UNC), Chapel Hill, NC, United States.
Department of Medicine, Thurston Arthritis Research Center, Division of Rheumatology, Allergy, and Immunology, University of North Carolina at Chapel Hill (UNC), Chapel Hill, NC, United States.
Front Immunol. 2022 Sep 29;13:974374. doi: 10.3389/fimmu.2022.974374. eCollection 2022.
10% of US residents have food allergies, including 2% with peanut allergy. Mast cell mediators released during the allergy effector phase drive allergic reactions. Therefore, targeting sensitized mast cells may prevent food allergy symptoms.
We used novel, human, allergen-specific, IgE monoclonal antibodies (mAbs) created using human hybridoma techniques to design an system to evaluate potential therapeutics targeting sensitized effector cells.
Two human IgE mAbs specific for peanut, generated through human hybridoma techniques, were used to sensitize rat basophilic leukemia (RBL) SX-38 cells expressing the human IgE receptor (FcϵRI). Beta-hexosaminidase release (a marker of degranulation), cytokine production, and phosphorylation of signal transduction proteins downstream of FcϵRI were measured after stimulation with peanut. Degranulation was also measured after engaging inhibitory receptors CD300a and Siglec-8.
Peanut-specific human IgE mAbs bound FcϵRI, triggering degranulation after stimulation with peanut in RBL SX-38 cells. Sensitized RBL SX-38 cells stimulated with peanut increased levels of phosphorylated SYK and ERK, signal transduction proteins downstream of FcϵRI. Engaging inhibitory cell surface receptors CD300a or Siglec-8 blunted peanut-specific activation.
Allergen-specific human IgE mAbs, expressed from human hybridomas and specific for a clinically relevant food allergen, passively sensitize allergy effector cells central to the models of the effector phase of food allergy. Peanut reproducibly activates and induces degranulation of RBL SX-38 cells sensitized with peanut-specific human IgE mAbs. This system provides a unique screening tool to assess the efficacy of therapeutics that target allergy effector cells and inhibit food allergen-induced effector cell activation.
美国有 10%的居民有食物过敏，包括 2%的花生过敏。过敏效应阶段释放的肥大细胞介质驱动过敏反应。因此，针对致敏的肥大细胞可能预防食物过敏症状。
我们使用新型、人类、过敏原特异性、IgE 单克隆抗体（mAbs），通过人类杂交瘤技术创建，设计了一个系统来评估针对致敏效应细胞的潜在治疗方法。
使用两种通过人类杂交瘤技术产生的针对花生的人类 IgE mAbs，使表达人类 IgE 受体（FcϵRI）的大鼠嗜碱性白血病（RBL）SX-38 细胞致敏。用花生刺激后，测量脱颗粒（脱粒的标志物）、细胞因子产生和 FcϵRI 下游信号转导蛋白的磷酸化。在用抑制性受体 CD300a 和 Siglec-8 结合后，也测量脱颗粒。
花生特异性人类 IgE mAbs 结合 FcϵRI，在 RBL SX-38 细胞中用花生刺激后触发脱粒。用花生刺激致敏的 RBL SX-38 细胞增加了 FcϵRI 下游信号转导蛋白 SYK 和 ERK 的磷酸化水平。结合抑制性细胞表面受体 CD300a 或 Siglec-8 可减弱花生特异性激活。
从人类杂交瘤表达的、针对临床相关食物过敏原的过敏原特异性人类 IgE mAbs，被动致敏过敏效应细胞，这些细胞是食物过敏效应阶段的 模型的核心。花生可重现地激活和诱导用花生特异性人类 IgE mAbs 致敏的 RBL SX-38 细胞脱颗粒。该系统提供了一个独特的筛选工具，用于评估针对过敏效应细胞并抑制食物过敏原诱导的效应细胞激活的治疗方法的疗效。
