University of Texas MD Anderson Cancer Center

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Mass Spectrometric Analysis of Glycosphingolipid Antigens

Alexandra Bili Yin¹, David Hawke², 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.

Immunology and Infection

Quantitative High-throughput Single-cell Cytotoxicity Assay For T Cells

Ivan Liadi¹, Jason Roszik², Gabrielle Romain¹, Laurence J.N. Cooper², Navin Varadarajan¹

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.

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 Huls¹, Matthew J. Figliola¹, Margaret J. Dawson¹, Simon Olivares¹, Partow Kebriaei², Elizabeth J. Shpall², Richard E. Champlin², Harjeet Singh¹, Laurence J.N. Cooper¹

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.

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. Moyes¹, M. Helen Huls¹, Priti Tewari¹, Elizabeth J. Shpall², Richard Champlin², Laurence J.N. Cooper¹

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.