抗人卵巢癌/抗人CD3单链双特异性抗体介导的αβT细胞CDR3谱系漂移

目的探讨抗人卵巢癌/抗人CD3单链双特异性抗体(BHL-1)介导的αβT细胞CDR3谱系漂移,为双特异性抗体介导的T细胞免疫应答机制提供理论基础。方法采用免疫扫描谱型分析技术,分析6例正常献血员T细胞在人卵巢癌SKOV3细胞联合BHL-1刺激前后的TCR库多样性变化(CDR3谱型分布特征)及刺激后T细胞TCRα、β链CDR3优势利用情况,对克隆性增生T细胞的CDR3区进行序列分析。结果刺激前6例正常献血员TCR CDR3谱型均呈高斯分布,刺激后发生CDR3谱系漂移,部分TCR Vα、Vβ家族出现优势增生,明确了BHL-1介导下单克隆增生T细胞的α、β链CDR3序列。结论 SKOV3联合BHL-1诱导的T细胞CDR3谱系出现明显漂移,提示CDR3的选择性表达可能与BHL-1介导的特异性T细胞免疫反应有关,特异应答T细胞TCR CDR3序列的确定,将为卵巢癌的T细胞免疫治疗提供基础。

An anti-human ovarian carcinoma and CD3 bispecific single-chain antibody mediates CDR3 spectratype drift of T cell receptor alpha and beta chains

Objective To analyze the drift of T cell receptor(TCR) Vα and Vβ gene family CDR3 spectratype in response to ovarian carcinoma cells mediated by an anti-human ovarian carcinoma/CD3 bispecific single-chain antibody(BHL-1),and explore the mechanism of the bispecific single-chain antibody-mediated T cell immune response.Methods Immunoscopic spectratyping technique was used to analyze the TCR repertoire diversity(CDR3 spectratype distribution) of the T cells from 6 healthy donors before and after stimulation of the cells with human ovarian carcinoma in the presence of BHL-1.The predominant usage of TCR α and β chain CDR3 was analyzed after the stimulation,and sequence analysis was performed for the CDR3 region of the monoclonal T cells.Results The spectratypes of Vα and Vβ gene family TCR CDR3 region showed a Gaussian distribution before stimulation of the T cells from the 6 donors.After stimulation of the T cells,CDR3 spectratype drift occurred in the T cells,and some TCR Vα and Vβ families showed an anomalous and oligoclonal expansion.Different CDR3 sequences of the Vα and Vβ gene family TCR were found in the monoclonal T cells stimulated with BHL-1.Conclusion CDR3 spectratype drift occurs in TCR α and β chains of T cells after stimulation with human ovarian carcinoma cells and BHL-1,indicating that the predominant usage of TCR Vα and Vβ families is associated with the specific T cell immune response mediated by BHL-1.