Jeon Ji-Ho, Hong Cheol-Hwa, Hyun You-Seok, Lee Hyeyoung, Oh Eun-Jee, Kim Tai-Gyu, Baek In-Cheol
Catholic Hematopoietic Stem Cell Bank, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
Department of Microbiology, College of Medicine, The Catholic University of Korea, Seoul, Republic of Korea.
BMC Mol Cell Biol. 2025 Jul 16;26(1):22. doi: 10.1186/s12860-025-00549-5.
Antibodies against non-HLA antigens, such as MICA and MICB, have emerged as potential contributors to antibody-mediated rejection and graft failure. While MICA antibodies are well characterized, MICB-specific antibodies remain poorly understood due to the lack of standardized detection tools. To address this gap, we aimed to develop a cell-based platform expressing individual MICB antigens to evaluate the feasibility of detecting allele-specific anti-MICB antibodies in pre-kidney transplant sera.
HLA class I, MICA, and MICB-null human embryonic kidney (HEK)-293T cells were previously generated by CRISPR/Cas9. We established five cell lines expressing single MICB antigens (each MICB*002, *003, *004, *005:02, and *008 allele). A total of 64 pre-kidney transplant sera were tested to assess anti-MICB antibody responses to the five cell lines using flow cytometry.
We successfully established and validated five HEK-293T cell lines each expressing a single MICB antigen using anti-MICB monoclonal antibody staining. No anti-MICB antibodies were detected in any of the 64 pre-transplant sera tested. This finding may reflect a low incidence of sensitization to MICB in this patient population and suggests the need for larger, more diverse cohorts in future studies to fully assess the prevalence of anti-MICB responses. The established cell lines provide a promising tool for future investigation of allele-specific anti-MICB antibody responses.
While the present study did not detect allele-specific anti-MICB antibody responses, establishing HEK-293T cell lines expressing single MICB antigens represents a significant methodological advance. This platform enables the potential assessment of immune responses targeted to individual MICB allotypes, thus offering new avenues for the future study of MICB immunogenicity in transplantation settings.
针对非 HLA 抗原（如 MICA 和 MICB）的抗体已成为抗体介导的排斥反应和移植物功能衰竭的潜在因素。虽然 MICA 抗体已得到充分表征，但由于缺乏标准化检测工具，对 MICB 特异性抗体仍知之甚少。为填补这一空白，我们旨在开发一个表达单个 MICB 抗原的细胞平台，以评估在肾移植前血清中检测等位基因特异性抗 MICB 抗体的可行性。
先前通过 CRISPR/Cas9 技术构建了 HLA I 类、MICA 和 MICB 基因缺失的人胚肾（HEK）-293T 细胞。我们建立了五个表达单个 MICB 抗原的细胞系（分别为每个 MICB*002、*003、*004、005:02 和008 等位基因）。使用流式细胞术对总共 64 份肾移植前血清进行检测，以评估其对这五个细胞系的抗 MICB 抗体反应。
我们使用抗 MICB 单克隆抗体染色成功建立并验证了五个分别表达单个 MICB 抗原的 HEK-293T 细胞系。在所检测的 64 份移植前血清中均未检测到抗 MICB 抗体。这一发现可能反映了该患者群体中对 MICB 致敏的发生率较低，并表明未来研究需要更大、更多样化的队列，以全面评估抗 MICB 反应的患病率。所建立的细胞系为未来研究等位基因特异性抗 MICB 抗体反应提供了一个有前景的工具。
虽然本研究未检测到等位基因特异性抗 MICB 抗体反应，但建立表达单个 MICB 抗原的 HEK-293T 细胞系代表了一项重大的方法学进展。该平台能够潜在地评估针对个体 MICB 同种异型的免疫反应，从而为未来移植环境中 MICB 免疫原性的研究提供新途径。
