Division of Gene Therapy and Regulation of Gene Expression, Cima Universidad de Navarra, Av. Pio XII 55, 31008, Pamplona, Spain.
Instituto de Investigación Sanitaria de Navarra (IdISNA), Pamplona, Spain.
Sci Rep. 2021 Nov 2;11(1):21427. doi: 10.1038/s41598-021-00787-5.
A promising therapy for patients with B-cell lymphoma is based on vaccination with idiotype monoclonal antibodies (mAbs). Since idiotypes are different in each tumor, a personalized vaccine has to be produced for each patient. Expression of immunoglobulins with appropriate post-translational modifications for human use often requires the use of stable mammalian cells that can be scaled-up to reach the desired level of production. We have used a noncytopathic self-amplifying RNA vector derived from Semliki Forest virus (ncSFV) to generate BHK cell lines expressing murine follicular lymphoma-derived idiotype A20 mAb. ncSFV/BHK cell lines expressed approximately 2 mg/L/24 h of A20 mAb with proper quaternary structure and a glycosylation pattern similar to that of A20 mAb produced by hybridoma cells. A20 mAb purified from the supernatant of a ncSFV cell line, or from the hybridoma, was conjugated to keyhole limpet hemocyanin and used to immunize Balb/c mice by administration of four weekly doses of 25 µg of mAb. Both idiotype mAbs were able to induce a similar antitumor protection and longer survival compared to non-immunized mice. These results indicate that the ncSFV RNA vector could represent a quick and efficient system to produce patient-specific idiotypes with potential application as lymphoma vaccines.
用于治疗 B 细胞淋巴瘤患者的一种有前景的疗法是基于接种独特型单克隆抗体 (mAb) 的疫苗。由于每个肿瘤中的独特型都不同,因此必须为每个患者生产个性化的疫苗。为了实现人类使用的适当翻译后修饰的免疫球蛋白的表达,通常需要使用可规模化达到所需生产水平的稳定哺乳动物细胞。我们使用源自 Semliki Forest 病毒 (ncSFV) 的非细胞病变性自我扩增 RNA 载体来生成表达源自鼠滤泡性淋巴瘤的独特型 A20 mAb 的 BHK 细胞系。ncSFV/BHK 细胞系以适当的四级结构和类似于杂交瘤细胞产生的 A20 mAb 的糖基化模式表达约 2mg/L/24h 的 A20 mAb。从 ncSFV 细胞系的上清液或杂交瘤中纯化的 A20 mAb 与钥孔贻贝血蓝蛋白缀合,并通过每周给予 25μg mAb 的四剂免疫 Balb/c 小鼠。两种独特型 mAb 都能够诱导类似的抗肿瘤保护和更长的生存时间,与未免疫的小鼠相比。这些结果表明,ncSFV RNA 载体可以代表一种快速有效的系统,用于生产具有作为淋巴瘤疫苗潜在应用的患者特异性独特型。