Fung-Leung W P, De Sousa-Hitzler J, Ishaque A, Zhou L, Pang J, Ngo K, Panakos J A, Chourmouzis E, Liu F T, Lau C Y
R.W. Johnson Pharmaceutical Research Institute (La Jolla), San Diego, California 92121, USA.
J Exp Med. 1996 Jan 1;183(1):49-56. doi: 10.1084/jem.183.1.49.
The high-affinity receptor for immunoglobulin (Ig) E (Fc epsilon RI) on mast cells and basophils plays a key role in IgE-mediated allergies. Fc epsilon RI is composed of one alpha, one beta, and two gamma chains, which are all required for cell surface expression of Fc epsilon RI, but only the alpha chain is involved in the binding to IgE. Fc epsilon RI-IgE interaction is highly species specific, and rodent Fc epsilon RI does not bind human IgE. To obtain a "humanized" animal model that responds to human IgE in allergic reactions, transgenic mice expressing the human Fc epsilon RI alpha chain were generated. The human Fc epsilon RI alpha chain gene with a 1.3-kb promoter region as a transgene was found to be sufficient for mast cell-specific transcription. Cell surface expression of the human Fc epsilon RI alpha chain was indicated by the specific binding of human IgE to mast cells from transgenic mice in flow cytometric analyses. Expression of the transgenic Fc epsilon RI on bone marrow-derived mast cells was 4.7 x 10(4)/cell, and the human IgE-binding affinity was Kd = 6.4 nM in receptor-binding studies using 125I-IgE. The transgenic human Fc epsilon RI alpha chain was complexed with the mouse beta and gamma chains in immunoprecipitation studies. Cross-linking of the transgenic Fc epsilon RI with human IgE and antigens led to mast cell activation as indicated by enhanced tyrosine phosphorylation of the Fc epsilon RI beta and gamma chains and other cellular proteins. Mast cell degranulation in transgenic mice could be triggered by human IgE and antigens, as demonstrated by beta-hexosaminidase release in vitro and passive cutaneous anaphylaxis in vivo. The results demonstrate that the human Fc epsilon RI alpha chain alone not only confers the specificity in human IgE binding, but also can reconstitute a functional receptor by coupling with the mouse beta and gamma chains to trigger mast cell activation and degranulation in a whole animal system. These transgenic mice "humanized" in IgE-mediated allergies may be valuable for development of therapeutic agents that target the binding of IgE to its receptor.
肥大细胞和嗜碱性粒细胞上的免疫球蛋白(Ig)E高亲和力受体(FcεRI)在IgE介导的过敏反应中起关键作用。FcεRI由一条α链、一条β链和两条γ链组成,这些链对于FcεRI在细胞表面的表达都是必需的,但只有α链参与与IgE的结合。FcεRI与IgE的相互作用具有高度的物种特异性,啮齿动物的FcεRI不与人IgE结合。为了获得在过敏反应中对人IgE有反应的“人源化”动物模型,制备了表达人FcεRIα链的转基因小鼠。发现带有1.3 kb启动子区域的人FcεRIα链基因作为转基因足以进行肥大细胞特异性转录。流式细胞术分析表明人IgE与转基因小鼠肥大细胞的特异性结合表明了人FcεRIα链在细胞表面的表达。在骨髓来源的肥大细胞上转基因FcεRI的表达为4.7×10⁴/细胞,在使用¹²⁵I-IgE的受体结合研究中,人IgE结合亲和力为Kd = 6.4 nM。在免疫沉淀研究中,转基因人FcεRIα链与小鼠β链和γ链复合。转基因FcεRI与人IgE和抗原的交联导致肥大细胞活化,表现为FcεRIβ链和γ链以及其他细胞蛋白酪氨酸磷酸化增强。如体外β-己糖胺酶释放和体内被动皮肤过敏反应所示,人IgE和抗原可触发转基因小鼠的肥大细胞脱颗粒。结果表明,单独的人FcεRIα链不仅赋予人IgE结合特异性,还可以通过与小鼠β链和γ链偶联来重建功能性受体,从而在整个动物系统中触发肥大细胞活化和脱颗粒。这些在IgE介导的过敏反应中“人源化”的转基因小鼠对于开发针对IgE与其受体结合的治疗药物可能具有重要价值。