%0 Generic %A Simon, Sylvain %A Wu, Zhong %A Cruard, J. %A Vignard, Virginie %A Fortun, Agnes %A Khammari, Amir %A Dreno, Brigitte %A Lang, Francois %A Rulli, Samuel J. %A Labarriere, Nathalie %D 2018 %T Data_Sheet_3_TCR Analyses of Two Vast and Shared Melanoma Antigen-Specific T Cell Repertoires: Common and Specific Features.PDF %U https://frontiersin.figshare.com/articles/dataset/Data_Sheet_3_TCR_Analyses_of_Two_Vast_and_Shared_Melanoma_Antigen-Specific_T_Cell_Repertoires_Common_and_Specific_Features_PDF/7027703 %R 10.3389/fimmu.2018.01962.s003 %2 https://frontiersin.figshare.com/ndownloader/files/12912500 %K TCR sequencing %K melanoma %K Melan-A %K MELOE-1 %K immunotherapy %X

Among Immunotherapeutic approaches for cancer treatment, the adoptive transfer of antigen specific T cells is still a relevant approach, that could have higher efficacy when further combined with immune check-point blockade. A high number of adoptive transfer trials have been performed in metastatic melanoma, due to its high immunogenic potential, either with polyclonal TIL or antigen-specific polyclonal populations. In this setting, the extensive characterization of T cell functions and receptor diversity of infused polyclonal T cells is required, notably for monitoring purposes. We developed a clinical grade procedure for the selection and amplification of polyclonal CD8 T cells, specific for two shared and widely expressed melanoma antigens: Melan-A and MELOE-1. This procedure is currently used in a clinical trial for HLA-A2 metastatic melanoma patients. In this study, we characterized the T-cell diversity (T-cell repertoire) of such T cell populations using a new RNAseq strategy. We first assessed the added-value of TCR receptor sequencing, in terms of sensitivity and specificity, by direct comparison with cytometry analysis of the T cell populations labeled with anti-Vß-specific antibodies. Results from these analyzes also confirmed specific features already reported for Melan-A and MELOE-1 specific T cell repertoires in terms of V-alpha recurrence usage, on a very high number of T cell clonotypes. Furthermore, these analyses also revealed undescribed features, such as the recurrence of a specific motif in the CDR3α region for MELOE-1 specific T cell repertoire. Finally, the analysis of a large number of T cell clonotypes originating from various patients revealed the existence of public CDR3α and ß clonotypes for Melan-A and MELOE-1 specific T cells. In conclusion, this method of high throughput TCR sequencing is a reliable and powerful approach to deeply characterize polyclonal T cell repertoires, and to reveal specific features of a given TCR repertoire, that would be useful for immune follow-up of cancer patients treated by immunotherapeutic approaches.

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