<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "Generation of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy"^^ . "Adoptive cell therapy using tumor antigen-specific CD8+ T cells is a promising approach to treat\r\npatients with advanced or metastasizing cancer including malignant melanoma. The general procedure\r\nis based on the isolation, in vitro stimulation and expansion of tumor-infiltrating lymphocytes\r\n(TILs). These activated and expanded TILs, which are specific for common tumor antigens are then\r\ntransferred back into the patients to attack and neutralize cancer cells. However, current studies report\r\ndisease relapse in treated patients because of T cell exhaustion after ongoing TCR stimulation\r\nresulting in a decreased effector function and a loss of their replicative capacity.\r\nThe use of induced pluripotent stem cells (iPSCs) as a source for the generation of “off-the-shelf”\r\nCD8+ T cells could enable T cell-based immunotherapy on a large scale. Similar to embryonic stem\r\ncells, iPSCs have the potential to give rise to nearly unlimited amounts of any cell type, including\r\nCD8+ T cells. The aim of this project was to develop strategies for the in vitro generation of\r\nmelanoma antigen-specific CD8+ T cells from human iPSCs.\r\nFirst, iPSCs were transfected with S/MAR DNA vectors coding for a T cell receptor (TCR) against\r\nMART-1 or a chimeric antigen receptor (CAR) directed against MCSP. S/MAR DNA vectors are\r\neffective gene delivery systems since they contain a scaffold/matrix attachment region, which binds\r\nto the nuclear matrix and ensures a persistent gene expression over hundreds of cell divisions. The\r\ntransfection of iPSCs with the S/MAR DNA vector did not alter the gene expression of the pluripotency\r\ngenes NANOG and OCT4 as well as other T cell-related differentiation genes.\r\nNext, a 2D co-culture system with OP9 murine stromal cells was used to facilitate the differentiation\r\nof CD34+ hematopoietic stem and progenitors cells (HSPCs) from iPSCs. Further differentiation\r\nof CD34+ HSPCs into CD4+CD8+ double-positive (DP) T cells was achieved by long-term coculture\r\nwith bioengineered OP9 stromal cells expressing T cell-specific cytokines like FLT3-ligand,\r\nCXCL12 and SCF as well as the Notch ligand DLL4 and supplementation of the growth medium\r\nwith recombinant IL7.\r\nSince clinical applications require clean populations of effector T cells without contamination with\r\nmurine cells, the stromal cell based co-culture system was replaced with a commercially available\r\ndifferentiation kit. Following the manufacturer’s instructions, I was able to generate a limited\r\namount of CD8+ single-positive (SP) T cells. Flow cytometric analysis of common T cell markers\r\nrevealed a low CD3 expression in generated T cells but a persistent MART-1 TCR or MCSP CAR\r\nexpression, respectively. In comparison to CD4+CD8+ DP T cells, CD8+ SP T cells demonstrated\r\nthe capability to produce cytokines, showed degranulation capacity after stimulation and exhibited\r\ncytotoxic effects against melanoma cell lines.\r\nHence, these findings revealed the possibilities as well as the challenges connected with the use of\r\nhuman iPSCs for the generation of CD8+ T cells. However, the use of S/MAR DNA vectors and\r\nnovel differentiation strategies could enable further clinical applications in cancer treatment and\r\nhelp to improve common immunotherapies for melanoma patients."^^ . "2024" . . . . . . . "Juliane"^^ . "Poelchen"^^ . "Juliane Poelchen"^^ . . . . . . "Generation of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy (PDF)"^^ . . . "Thesis_Juliane Poelchen-1.pdf"^^ . . . "Generation of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy (Other)"^^ . . . . . . "indexcodes.txt"^^ . . . "Generation of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "Generation of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy (Other)"^^ . . . . . . "preview.jpg"^^ . . . "Generation of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy (Other)"^^ . . . . . . "medium.jpg"^^ . . . "Generation of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy (Other)"^^ . . . . . . "small.jpg"^^ . . "HTML Summary of #33373 \n\nGeneration of melanoma antigen-specific CD8+ T cells from human induced pluripotent stem cells for adoptive cell therapy\n\n" . "text/html" . . . "570 Biowissenschaften, Biologie"@de . "570 Life sciences"@en . .