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University of Texas MD Anderson Cancer Center

4 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Mass Spectrometric Analysis of Glycosphingolipid Antigens
Alexandra Bili Yin 1, David Hawke 2, Dapeng Zhou 3,4
1Undergraduate Program, Rice University , 2Proteomics Facility, Department of Pathology, University of Texas MD Anderson Cancer Center , 3Department of Melanoma Medical Oncology, University of Texas MD Anderson Cancer Center , 4University of Texas Graduate School of Biological Sciences at Houston

A specific and sensitive method to gain insight into the expression profile of glycosphingolipid antigens in immune organs and cells is described. The method takes advantage of the ion trap mass spectrometry allowing step-wise fragmentation of glycosphingolipid molecules for structural analysis in comparison to chemically synthesized standards.

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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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