U.T. MD Anderson Cancer Center

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Expanding Cytotoxic T Lymphocytes from Umbilical Cord Blood that Target Cytomegalovirus, Epstein-Barr Virus, and Adenovirus

Patrick J. Hanley¹, Sharon Lam^1,2, Elizabeth J. Shpall³, 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.

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.