GENE THERAPY WITH MAB DRIVATIVES EXPRESSED ON TUMORS

使用在肿瘤上表达的 MAB 驱动程序进行基因治疗

• 批准号: 6923079
• 负责人: JEFFREY A LEDBETTER
• 金额: $ 31.5万
• 依托单位: TRUBION PHARMACEUTICALS, INC.
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-06-03 至 2005-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6923079
• 关键词: CD antigens; T cell receptor; T lymphocyte; animal tissue; antigen antibody reaction; antireceptor antibody; antitumor antibody; biological signal transduction; cytotoxic T lymphocyte; flow cytometry; gene expression; gene therapy; helper T lymphocyte; immunoglobulin genes; immunologic assay /test; leukocyte activation /transformation; monoclonal antibody; neoplasm /cancer genetics; neoplasm /cancer therapy; neoplastic cell; receptor expression; tissue /cell culture; transfection /expression vector; tumor antigens

DESCRIPTION: (Applicant's Abstract) Previous work in the applicant's laboratory has demonstrated that costimulation plays a key role in the induction of an immune response, including one to tumors. This costimulation can be provided by either transfecting a gene encoding a ligand, such as CD80 and/or CD86 into tumor cells, or by transfecting a gene encoding a single chain Fv (scFv) from the appropriate anti-receptor mAb. The scFv approach can have an advantage over the use of natural ligands. For example, CD80 and CD86 bind with high avidity to CTLA-4, which produces a strong negative signal, while the anti-CD28 scFv does not. In addition, scFvs can be used when natural ligands are unknown, such as for CD3-epsilon. In this application, variable region genes from hybridoma antibodies specific for human costimulatory receptors will be expressed as cell surface scFvs to modulate antigen-specific immune responses to tumor cells. Genes encoding scFvs that retain specificity and high binding affinity for CD3-epsilon, CD28, CD2, CD4, CD8, and CD154 have been constructed and expressed as soluble Ig-fusion proteins. Transmembrane domains have been added to some of the scFv gene constructs to direct their expression to the cell surface. These genes will be expressed on the surface of human tumor cell lines to determine their potential for amplification of T cell responses. The applicant hypothesizes that use of antibody derivatives can improve the specificity and potency of cancer gene therapy, and that cell surface expression of scFvs will enhance the immune response to specific antigens when compared with expression of native ligands for T cell surface receptors. He will determine whether expression of scFvs specific for T cell stimulatory and costimulatory molecules on the surface of tumor cells will increase T cell activation, including both CD4+ Th1 responses and CD8+ cytotoxic responses important for tumor rejection.

描述：（申请人的摘要）申请人实验室的先前工作 已经证明，共刺激在诱导中起着关键作用 免疫反应，包括对肿瘤的一种。可以由 要么转染编码配体的基因，例如CD80和/或CD86 肿瘤细胞，或通过转染从编码单链FV（SCFV）的基因 适当的抗受体mAb。 SCFV方法可以具有优势 天然配体的使用。例如，CD80和CD86与高潮结合 到CTLA-4产生强信号的CTLA-4，而抗CD28 SCFV 没有。此外，当天然配体未知时，可以使用SCFV，这样 至于CD3- epsilon。在此应用中，杂交瘤的可变区域基因 针对人类共刺激受体特异的抗体将表示为细胞 表面SCFV调节对肿瘤细胞的抗原特异性免疫反应。 编码保留特异性和高结合亲和力的SCFV的基因 CD3- EPSILON，CD28，CD2，CD4，CD8和CD154已构造并表达 作为可溶性IG融合蛋白。跨膜域已添加到其中一些 SCFV基因构建以将其表达引导到细胞表面。这些 基因将在人类肿瘤细胞系的表面表达 它们扩增T细胞反应的潜力。申请人 假设使用抗体衍生物可以提高特异性和 癌症基因疗法的效力以及SCFV的细胞表面表达将 与表达相比，增强对特定抗原的免疫反应 T细胞表面受体的天然配体。他将确定是否 对T细胞刺激和共刺激分子的SCFV的表达 在肿瘤细胞的表面将增加T细胞的激活，包括 CD4+ TH1反应和CD8+细胞毒性反应对于肿瘤排斥很重要。

项目成果

Expression and characterization of recombinant soluble human CD3 molecules: presentation of antigenic epitopes defined on the native TCR-CD3 complex.

重组可溶性人 CD3 分子的表达和表征：天然 TCR-CD3 复合物上定义的抗原表位的呈现。

• DOI: 10.1093/intimm/14.4.389
• 发表时间: 2002
• 期刊: International immunology
• 影响因子: 4.4
• 作者: Law,Che-Leung;Hayden-Ledbetter,Martha;Buckwalter,Sonya;McNeill,Lisa;Nguyen,Hieu;Habecker,Phil;Thorne,BarbaraA;Dua,Raj;Ledbetter,JeffreyA
• 通讯作者: Ledbetter,JeffreyA