Yamazaki Tatsuya, Iwasaki Kenta, Tomono Susumu, Imai Masaki, Miwa Yuko, Shizuku Masato, Ashimine Satoshi, Ishiyama Kohei, Inui Masanori, Okuzaki Daisuke, Okada Manabu, Kobayashi Takaaki, Akashi-Takamura Sachiko
Department of Microbiology and Immunology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
Department of Kidney Diseases and Transplant Immunology, Aichi Medical University School of Medicine, Nagakute, Aichi, Japan.
iScience. 2024 Aug 3;27(9):110649. doi: 10.1016/j.isci.2024.110649. eCollection 2024 Sep 20.
Detecting antibodies, particularly those targeting donor human leukocyte antigens in organ transplantation and self-antigens in autoimmune diseases, is crucial for diagnosis and therapy. Radioprotective 105 (RP105), a Toll-like receptor family protein, is expressed in immune-competent cells, such as B cells. Studies in mice have shown that the anti-mouse RP105 antibody strongly activates B cells and triggers an adjuvant effect against viral infections. However, the anti-human RP105 antibody (ɑhRP105) weakly activates human B cells. This study established new culture conditions under, which human B cells are strongly activated by the ɑhRP105. When combined with CpGDNA, specific antibody production against blood group carbohydrates, ɑGal, and SARS-CoV-2 was successfully detected in human B cell cultures. Furthermore, comprehensive analysis using liquid chromatography-electrospray ionization tandem mass spectrometry, single-cell RNA sequencing, and quantitative real-time PCR revealed that ɑhRP105 triggered a different activation stimulus compared to CpGDNA. These findings could help identify antibody-producing B cells in cases of transplant rejection and autoimmune diseases.
检测抗体,尤其是那些针对器官移植中供体人类白细胞抗原和自身免疫性疾病中自身抗原的抗体,对于诊断和治疗至关重要。辐射防护蛋白105(RP105)是一种Toll样受体家族蛋白,在免疫活性细胞如B细胞中表达。对小鼠的研究表明,抗小鼠RP105抗体能强烈激活B细胞并引发针对病毒感染的佐剂效应。然而,抗人RP105抗体(ɑhRP105)对人B细胞的激活作用较弱。本研究建立了新的培养条件,在此条件下ɑhRP105能强烈激活人B细胞。当与CpGDNA联合使用时,在人B细胞培养物中成功检测到针对血型碳水化合物、α半乳糖和严重急性呼吸综合征冠状病毒2(SARS-CoV-2)的特异性抗体产生。此外,使用液相色谱-电喷雾电离串联质谱、单细胞RNA测序和定量实时聚合酶链反应进行的综合分析表明,与CpGDNA相比,ɑhRP105引发了不同的激活刺激。这些发现有助于在移植排斥和自身免疫性疾病病例中识别产生抗体的B细胞。