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U.T. MD Anderson Cancer Center

4 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Expanding Cytotoxic T Lymphocytes from Umbilical Cord Blood that Target Cytomegalovirus, Epstein-Barr Virus, and Adenovirus
Patrick J. Hanley 1, Sharon Lam 1,2, Elizabeth J. Shpall 3, Catherine M. Bollard 1,2,4,5
1Center for Cell and Gene Therapy, Baylor College of Medicine , 2Pathology and Immunology, Baylor College of Medicine , 3Department of Stem Cell Transplantation and Cellular Therapy, University of Texas M.D. Anderson Cancer Center, 4Medicine, Baylor College of Medicine , 5Department of Pediatrics, Baylor College of Medicine

Here we describe the first good manufacturing practice (GMP)-compliant method of producing virus-specific cytotoxic T lymphocytes (CTL) from umbilical cord blood, a source of predominantly naîve T cells.

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Immunology and Infection

Quantitative High-throughput Single-cell Cytotoxicity Assay For T Cells
Ivan Liadi 1, Jason Roszik 2, Gabrielle Romain 1, Laurence J.N. Cooper 2, Navin Varadarajan 1
1Department of Chemical and Biomolecular Engineering, University of Houston , 2Division of Pediatrics, Research Unit 907, University of Texas MD Anderson Cancer Center

We describe a single-cell high-throughput assay to measure cytotoxicity of T cells when incubated with tumor target cells. This method employs a dense, elastomeric array of sub-nanoliter wells (~100,000 wells/array) to spatially confine the T cells and target cells at defined ratios and is coupled to fluorescence microscopy to monitor effector-target conjugation and subsequent apoptosis.

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Immunology and Infection

Clinical Application of Sleeping Beauty and Artificial Antigen Presenting Cells to Genetically Modify T Cells from Peripheral and Umbilical Cord Blood
M. Helen Huls 1, Matthew J. Figliola 1, Margaret J. Dawson 1, Simon Olivares 1, Partow Kebriaei 2, Elizabeth J. Shpall 2, Richard E. Champlin 2, Harjeet Singh 1, Laurence J.N. Cooper 1
1Division of Pediatrics, U.T. MD Anderson Cancer Center, 2Department of Stem Cell Transplantation and Cellular Therapy, U.T. MD Anderson Cancer Center

T cells expressing a CD19-specific chimeric antigen receptor (CAR) are infused as investigational treatment of B-cell malignancies in our first-in-human gene therapy trials. We describe genetic modification of T cells using the Sleeping Beauty (SB) system to introduce CD19-specific CAR and selective propagation on designer CD19+ artificial antigen presenting cells.

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Immunology and Infection

Automated Cell Enrichment of Cytomegalovirus-specific T cells for Clinical Applications using the Cytokine-capture System
Pappanaicken Kumaresan *1, Mathew Figliola *1, Judy S. Moyes 1, M. Helen Huls 1, Priti Tewari 1, Elizabeth J. Shpall 2, Richard Champlin 2, Laurence J.N. Cooper 1
1Division of Pediatrics, U.T. MD Anderson Cancer Center, 2Stem Cell Transplantation and Cellular Therapy, U.T. MD Anderson Cancer Center

The goal of this protocol is to manufacture pathogen-specific clinical-grade T cells using a bench-top, automated, second generation cell enrichment device that incorporates a closed cytokine capture system and does not require dedicated staff or use of a GMP facility. The cytomegalovirus pp65-specific-T cells generated can be directly administered to patients.

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