Department I of Internal Medicine, University Hospital Cologne, Faculty of Medicine, University of Cologne, 50937 Cologne, Germany.
Center for Molecular Medicine Cologne (CMMC), University Hospital Cologne, Faculty of Medicine, University of Cologne, 50937 Cologne, Germany.
Cells. 2023 Dec 21;13(1):20. doi: 10.3390/cells13010020.
Antiviral neutralizing antibodies (nAbs) are commonly derived from B cells developed in immunized or infected animals and humans. Fully human antibodies are preferred for clinical use as they are potentially less immunogenic. However, the function of B cells varies depending on their homing pattern and an additional hurdle for antibody discovery in humans is the source of human tissues with an immunological microenvironment. Here, we show an efficient method to pharm human antibodies using immortalized B cells recovered from Nod.Rag.Gamma (NRG) mice reconstituting the human immune system (HIS). Humanized HIS mice were immunized either with autologous engineered dendritic cells expressing the human cytomegalovirus gB envelope protein (HCMV-gB) or with Epstein-Barr virus-like particles (EB-VLP). Human B cells recovered from spleen of HIS mice were efficiently immortalized with EBV in vitro. We show that these immortalized B cells secreted human IgGs with neutralization capacities against prototypic HCMV-gB and EBV-gp350. Taken together, we show that HIS mice can be successfully used for the generation and pharming fully human IgGs. This technology can be further explored to generate antibodies against emerging infections for diagnostic or therapeutic purposes.
抗病毒中和抗体(nAbs)通常源自免疫或感染动物和人类中产生的 B 细胞。完全人源抗体因其潜在的免疫原性较低而更适合临床应用。然而,B 细胞的功能因归巢模式而异,人类抗体发现的另一个障碍是具有免疫微环境的人类组织来源。在这里,我们展示了一种使用从重建人类免疫系统(HIS)的 Nod.Rag.Gamma(NRG)小鼠中回收的永生化 B 细胞来制备人源抗体的有效方法。人源化 HIS 小鼠通过用表达人巨细胞病毒 gB 包膜蛋白(HCMV-gB)的自体工程化树突状细胞或 Epstein-Barr 病毒样颗粒(EB-VLP)进行免疫接种。从 HIS 小鼠脾脏中回收的人 B 细胞在体外通过 EBV 有效地永生化。我们表明,这些永生化 B 细胞分泌的人 IgG 对典型的 HCMV-gB 和 EBV-gp350 具有中和能力。总之,我们表明 HIS 小鼠可成功用于生成和制备完全人源 IgG。该技术可进一步探索用于诊断或治疗新兴感染的抗体生成。