用于犬淋巴瘤治疗的犬抗犬 CD20 抗体的生成。

Generation of a canine anti-canine CD20 antibody for canine lymphoma treatment.

机构信息

Laboratory of Molecular Diagnostics and Therapeutics, The United Graduate School of Veterinary Medicine, Yamaguchi University, 1677-1 Yoshida, Yamaguchi, Yamaguchi Prefecture, 753-8515, Japan.

Department of Antibody Drug Development, Tohoku University Graduate School of Medicine, Miyagi, Japan.

出版信息

Sci Rep. 2020 Jul 10;10(1):11476. doi: 10.1038/s41598-020-68470-9.

DOI:10.1038/s41598-020-68470-9

PMID:32651429

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7351721/

Abstract

Lymphoma is the most common hematological cancer in dogs. Canine diffuse large B cell lymphoma shows a relatively good response to treatment with multi-agent cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP) chemotherapy; however, the 2-year survival rate is as low as 20%. For human B cell type lymphoma, the anti-CD20 chimeric antibody, rituximab, was developed two decades ago. The combination of rituximab and CHOP chemotherapy was highly successful in improving patient prognosis. However, no anti-canine CD20 antibody is available for the treatment of canine lymphoma. During this study, a rat anti-canine CD20 monoclonal antibody was established. We also generated a rat-canine chimeric antibody against canine CD20 designed for clinical application. This chimeric antibody (4E1-7-B) showed in vitro antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) against the canine B cell lymphoma cell line CLBL-1. Moreover, to obtain stronger ADCC activity, a defucosylated 4E1-7-B antibody (4E1-7-B_f) was also generated, and it showed tenfold stronger ADCC activity compared with 4E1-7-B. 4E1-7-B_f as well as 4E1-7-B suppressed the growth of CLBL-1 tumors in an immunodeficient xenotransplant mouse model. Finally, a single administration of 4E1-7-B_f induced considerable peripheral B cell depletion in healthy beagles. Thus, 4E1-7-B_f is a good antibody drug candidate for canine B cell type lymphoma.

摘要

淋巴瘤是犬类最常见的血液系统恶性肿瘤。犬类弥漫性大 B 细胞淋巴瘤对多柔比星、环磷酰胺、长春新碱和泼尼松（CHOP）联合化疗有较好的反应，但 2 年生存率仍低至 20%。对于人类 B 细胞淋巴瘤，二十年前开发了抗 CD20 嵌合抗体利妥昔单抗。利妥昔单抗联合 CHOP 化疗在改善患者预后方面非常成功。然而，目前尚无针对犬淋巴瘤的抗犬 CD20 抗体。在本研究中，我们制备了一种抗犬 CD20 的大鼠单克隆抗体，同时还针对犬 CD20 设计了一种用于临床应用的大鼠-犬嵌合抗体。该嵌合抗体（4E1-7-B）对犬 B 细胞淋巴瘤细胞系 CLBL-1 表现出体外抗体依赖性细胞介导的细胞毒性（ADCC）和补体依赖性细胞毒性（CDC）。此外，为了获得更强的 ADCC 活性，还制备了去岩藻糖基化的 4E1-7-B 抗体（4E1-7-B_f），其 ADCC 活性比 4E1-7-B 强 10 倍。4E1-7-B_f 和 4E1-7-B 在免疫缺陷性异种移植小鼠模型中均能抑制 CLBL-1 肿瘤的生长。最后，单次给予 4E1-7-B_f 可引起健康比格犬外周 B 细胞大量耗竭。因此，4E1-7-B_f 是犬 B 细胞淋巴瘤的一种有前景的抗体药物候选物。