在小鼠体内工程B细胞可产生高滴度的抗HIV广泛中和抗体(bNAb)，这一成果由以色列特拉维夫大学Adi Barzel小组经过不懈努力而取得。这一研究成果发表在2022年6月9日出版的国际学术期刊《自然-生物技术》上。

研究人员使用两种腺相关病毒载体对体内B细胞进行了改造， 一种编码金黄色葡萄球菌Cas9 (saCas9)，另一种编码3BNC117，一种抗HIV bNAb。将载体静脉注射到小鼠体内后，研究人员观察到由成功B细胞编辑诱导的记忆保留和bNAb分泌，中和滴度高达6.8 µg ml^-1。通过无偏倚CHANGE测序分析，研究人员检测到极低的CRISPR–Cas9 脱靶切割，而通过从B细胞特异性启动子表达saCas9减少了其它组织中靶向切割的产生。表达治疗性抗体的体内B细胞改造是一种安全、有效和可扩展的方法，它不仅适用于传染病，还适用于非传染性疾病的治疗，如癌症和自身免疫性疾病。

研究人员表示，移植体外工程化B细胞以分泌广泛中和抗体已在疾病模型中显示出疗效。然而，这种方法的临床转化需要专门的医疗中心、高技术要求的产生方式以及供体和受体细胞的主要组织相容性复合物具有相容性。

附：英文原文

Title: In vivo engineered B cells secrete high titers of broadly neutralizing anti-HIV antibodies in mice

Author: Nahmad, Alessio D., Lazzarotto, Cicera R., Zelikson, Natalie, Kustin, Talia, Tenuta, Mary, Huang, Deli, Reuveni, Inbal, Nataf, Daniel, Raviv, Yuval, Horovitz-Fried, Miriam, Dotan, Iris, Carmi, Yaron, Rosin-Arbesfeld, Rina, Nemazee, David, Voss, James E., Stern, Adi, Tsai, Shengdar Q., Barzel, Adi

Issue&Volume: 2022-06-09

Abstract: Transplantation of B cells engineered ex vivo to secrete broadly neutralizing antibodies (bNAbs) has shown efficacy in disease models. However, clinical translation of this approach would require specialized medical centers, technically demanding protocols and major histocompatibility complex compatibility of donor cells and recipients. Here we report in vivo B cell engineering using two adeno-associated viral vectors, with one coding for Staphylococcus aureus Cas9 (saCas9) and the other for 3BNC117, an anti-HIV bNAb. After intravenously injecting the vectors into mice, we observe successful editing of B cells leading to memory retention and bNAb secretion at neutralizing titers of up to 6.8μgml1. We observed minimal clustered regularly interspaced palindromic repeats (CRISPR)–Cas9 off-target cleavage as detected by unbiased CHANGE-sequencing analysis, whereas on-target cleavage in undesired tissues is reduced by expressing saCas9 from a B cell-specific promoter. In vivo B cell engineering to express therapeutic antibodies is a safe, potent and scalable method, which may be applicable not only to infectious diseases but also in the treatment of noncommunicable conditions, such as cancer and autoimmune disease.

DOI: 10.1038/s41587-022-01328-9

Source: https://www.nature.com/articles/s41587-022-01328-9