دورية أكاديمية
Single cell sequencing to identify TCRs that recognize autologous tumor cells after vaccination with allogeinic DRibble vaccine
| العنوان: | Single cell sequencing to identify TCRs that recognize autologous tumor cells after vaccination with allogeinic DRibble vaccine |
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| المؤلفون: | Hu, Hong-Ming, Paustian, Christopher C., Wen, Zhifa, Moudgil, Tarsem L., Hilton, Traci L., Bookhardt, Sam, Yu, Guangjie, Tran, Eric, Rajamanickam, Venkatesh, Urba, Walter, Sanborn, Rachel E., Fox, Bernard A. |
| المصدر: | Society for Immunotherapy of Cancer 2018 Annual Meeting Posters |
| بيانات النشر: | Providence St. Joseph Health Digital Commons |
| سنة النشر: | 2018 |
| المجموعة: | Providence St. Joseph Health Digital Commons |
| مصطلحات موضوعية: | Cancer, immunotherapy, cancer vaccines, T cells, tumor infiltrating lymphocytes, TIL, T-cell receptor sequencing, TCRseq, Oncology |
| الوصف: | Background: Adoptive immunotherapy with tumor-specific TCR gene-modified T cells has the potential to eradicate bulky disease. Traditional methods of TCR identification require lengthy in vitro culture to generate clonal T-cell populations, which adds time and complexity to this promising therapy. Here we described a simplified and reliable method to identify TCRs by single cell TCR sequencing of cells sorted with antibodies against T-cell surface markers that are up-regulated only when they are stimulated with specific tumor cell antigens. Methods: A tumor-infiltrating lymphocyte (TIL) culture with T cells reactive against autologous tumor was generated from a brain metastasis of a patients with NSCLC. A panel of antibodies against T-cell surface antigens was screened to identify markers that are specifically up-regulated after stimulation with autologous tumors but not with related allogeneic tumor cells. Tumor-specific T cells were sorted from TIL with three suitable antibodies and expanded by a rapid expansion protocol. Expanded T cells were examined for their tumor-specificity and subjected to single cell TCR sequencing using the 10X genomic system. The top 10 TCRs were identified by bio-informatics approach and the corresponding alpha and beta chains were synthesized and cloned into a retroviral vector based on MSG backbone. PBMC from healthy donors were transduced with the retrovirus supernatant after activation. Tumor- reactivity of transduced T cells was determined after expansion in media supplemented with IL-2, IL-7, and IL-15. To identify tumor-specific TCRs in PBMC from the same patient after vaccination with allogeneic DRibbles, we also developed a protocol to expand tumor-specific T cells from PBMC with in vitro stimulation with DRibble- loaded PBMC. Results: We identity CD94, CD137(4-1BB), CD355 (CRTAM) as specific markers for antigen-specific activation of T-cells by autologous tumor cells, whereas other “check point” markers such as CTLA-4, PD-1, Tim3, CD39, CD103 were up-regulated by ... |
| نوع الوثيقة: | text |
| وصف الملف: | application/pdf |
| اللغة: | unknown |
| العلاقة: | https://digitalcommons.psjhealth.org/sitc2018/2Test; https://digitalcommons.psjhealth.org/cgi/viewcontent.cgi?article=1002&context=sitc2018Test |
| الإتاحة: | https://digitalcommons.psjhealth.org/sitc2018/2Test https://digitalcommons.psjhealth.org/cgi/viewcontent.cgi?article=1002&context=sitc2018Test |
| رقم الانضمام: | edsbas.21794BE7 |
| قاعدة البيانات: | BASE |
| الوصف غير متاح. |