Sun Chunli, Jiang Yuan, Liu Shujun, He Qilin, Han Chengyao, Su Dai, Ma Hao, Guo Xingyu, Zhang Yan, Li Fubin, Zhang Huihui
Shanghai Institute of Immunology, Faculty of Basic Medicine, Key Laboratory of Cell Differentiation and Apoptosis of Chinese Ministry of Education, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Center for Immune-Related Diseases at Shanghai Institute of Immunology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China.
Int J Mol Sci. 2025 Apr 12;26(8):3664. doi: 10.3390/ijms26083664.
Sheep red blood cells (SRBC) has a long history as a classical T-cell dependent (TD) antigen. Due to its cost-effectiveness, easy accessibility, and ability to elicit a robust antibody immune response, SRBC continues to be widely used in studies related with humoral immunity modulation, vaccine development, and immunoactivity/immunotoxicity testing of bioactive agents. However, detecting the relative affinity levels of SRBC-specific antibodies in SRBC-immunized animal models remains challenging. Using flow cytometry, we established a detection system capable of quickly and accurately assessing the SRBC-specific antibody relative affinity levels in humoral samples (e.g., serum, tissue fluid) of SRBC-immunized mouse models. We further validated this method using affinity maturation-deficient mice, demonstrating that this method can distinguish affinity levels of the antibodies from different samples. This approach is simple and efficient, providing an accurate and effective technological solution for research on mechanisms of humoral immunity, antibody affinity maturation, vaccine response, and immunoactivity/immunotoxicity testing.
绵羊红细胞（SRBC）作为一种经典的T细胞依赖性（TD）抗原已有很长的历史。由于其成本效益高、易于获取且能够引发强烈的抗体免疫反应，SRBC继续广泛应用于体液免疫调节、疫苗开发以及生物活性剂的免疫活性/免疫毒性测试等相关研究中。然而，在SRBC免疫的动物模型中检测SRBC特异性抗体的相对亲和力水平仍然具有挑战性。我们利用流式细胞术建立了一种检测系统，能够快速、准确地评估SRBC免疫小鼠模型体液样本（如血清、组织液）中SRBC特异性抗体的相对亲和力水平。我们进一步使用亲和力成熟缺陷小鼠验证了该方法，证明该方法可以区分不同样本中抗体的亲和力水平。这种方法简单高效，为体液免疫机制、抗体亲和力成熟、疫苗反应以及免疫活性/免疫毒性测试的研究提供了准确有效的技术解决方案。
