一种红细胞同种异体免疫动物模型的构建*

doi:10.3969/j.issn.1671-2587.2025.03.010

临床输血与检验 ›› 2025, Vol. 27 ›› Issue (3): 340-346.DOI: 10.3969/j.issn.1671-2587.2025.03.010

一种红细胞同种异体免疫动物模型的构建^*

李济洋, 李辰琰, 周欠欠, 詹林盛

军事医学研究院, 北京 100850

收稿日期:2025-02-14 发布日期:2025-06-23
通讯作者: 詹林盛,主要从事输血免疫学工作,（E-mail）amms91@126.com。共同通信作者周欠欠,主要从事输血免疫学工作,（E-mail）zhouqianvv521@126.com。
作者简介:李济洋,主要从事免疫学工作,（E-mail）3312818133@qq,com。
基金资助:
*本课题受国家自然科学基金项目（No.82470232）资助

A Novel Animal Model for Red Blood Cell Alloimmunization

LI Jiyang, LI Chenyan, ZHOU Qianqian, ZHAN Linsheng

Academy of Military Medical Sciences, Beijing 100850

Received:2025-02-14 Published:2025-06-23

摘要/Abstract

摘要： 目的本研究旨在通过抗原工程化策略构建一种新型红细胞同种异体免疫动物模型,为研究红细胞输注引发的免疫反应奠定模型基础。方法采用乙基碳二亚胺盐酸盐（EDC）偶联技术将模式抗原鸡卵清蛋白（OVA）定向修饰至C57BL/6J小鼠红细胞表面,制备抗原工程化红细胞（RBC-OVA）。通过连续输注RBC-OVA至同品系小鼠体内,结合双光子活体成像技术,动态追踪红细胞在肝、脾中的黏附与清除过程。结果实验结果显示,OVA抗原修饰效率达99.5%,且未对红细胞形态、溶血率及细胞膜表面（CD47、PS）造成显著损伤。模型小鼠经4次输注后,血清中抗OVA IgG抗体浓度显著升高,抗体可特异性结合RBC-OVA。再次输注OVA修饰红细胞时,其外周血回收率较对照组显著降低,且在肝、脾中迅速被清除。结论本模型成功模拟了临床红细胞同种异体免疫反应,为研究抗体介导的输血无效机制及开发靶向干预策略提供了高效实验平台。

关键词: 红细胞同种异体免疫, 动物模型, 抗原工程化, EDC偶联技术

Abstract: Objective To develop a novel animal model of erythrocyte alloimmunity via an antigen engineering strategy, thus providing a foundational platform for investigating the immune response triggered by erythrocyte transfusion. Methods The model antigen ovalbumin (OVA) was specifically modified onto the surface of C57BL/6J mouse erythrocytes using ethyl carbodiimide hydrochloride (EDC) coupling technology to prepare antigen-engineered erythrocytes (RBC-OVA). Mice of the same strain were continuously transfused with RBC-OVA, and the adhesion and clearance processes of erythrocytes in the liver and spleen were dynamically tracked using two-photon in vivo imaging technology. Results The OVA antigen modification efficiency reached 99.5%, with no significant impact on erythrocyte morphology, hemolysis rate, or cell membrane surface markers (CD47, PS). After four transfusions, the anti-OVA IgG antibody concentration in the serum of the model mice significantly increased, and the antibody specifically bound to RBC-OVA. Upon re-infusion of OVA-modified red blood cells, the peripheral blood recovery rate was significantly lower than that of the control group, with rapid clearance observed in the liver and spleen. Conclusion This model effectively mimics the clinical red blood cell alloimmune response and provides a robust experimental platform for investigating the mechanisms underlying antibody-mediated transfusion failure and for developing targeted intervention strategies.

Key words: RBC alloimmunization, Animal model, Antigen engineering, EDC conjugation

中图分类号:

R392

李济洋, 李辰琰, 周欠欠, 詹林盛. 一种红细胞同种异体免疫动物模型的构建^*[J]. 临床输血与检验, 2025, 27(3): 340-346.

LI Jiyang, LI Chenyan, ZHOU Qianqian, ZHAN Linsheng. A Novel Animal Model for Red Blood Cell Alloimmunization[J]. JOURNAL OF CLINICAL TRANSFUSION AND LABORATORY MEDICINE, 2025, 27(3): 340-346.

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一种红细胞同种异体免疫动物模型的构建^*

A Novel Animal Model for Red Blood Cell Alloimmunization

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