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Baylor College of Medicine

8 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Generation of Multivirus-specific T Cells to Prevent/treat Viral Infections after Allogeneic Hematopoietic Stem Cell Transplant
Ulrike Gerdemann 1, Juan F. Vera 1, Cliona M. Rooney 1, Ann M. Leen 1
1Center for Cell and Gene Therapy, Baylor College of Medicine

A rapid, simple and cost-effective protocol for the generation of donor-derived multivirus-specific CTLs (rCTL) for infusion to allogeneic hematopoietic stem cell transplant (HSCT) recipients at risk of developing CMV, Adv or EBV infections. This manufacturing process is GMP-compliant and should ensure the broader implementation of T-cell immunotherapy beyond specialized centers.

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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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Bioengineering

In vitro Assembly of Semi-artificial Molecular Machine and its Use for Detection of DNA Damage
Candace L. Minchew 1,2, Vladimir V. Didenko 1,2,3
1Neurosurgery, Baylor College of Medicine , 2Michael E. DeBakey Veterans Affairs Medical Center, 3Molecular & Cellular Biology, Baylor College of Medicine

We demonstrate the assembly and application of a molecular-scale device powered by a topoisomerase protein. The construct is a bio-molecular sensor which labels two major types of DNA breaks in tissue sections by attaching two different fluorophores to their ends.

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Medicine

Analytical Techniques for Assaying Nitric Oxide Bioactivity
Hong Jiang 1, Deepa Parthasarathy 1, Ashley C. Torregrossa 1, Asad Mian 2, Nathan S. Bryan 1
1Texas Therapeutics Institute, University of Texas Health Science Center at Houston , 2Deptartment of Pediatrics, Baylor College of Medicine

The endogenous production of nitric oxide (NO) regulates a wide variety of biological functions. It is becoming increasingly clear that disruption or dysregulation of NO based signaling is involved in many human diseases. Methods to quantify relevant NO metabolites may provide novel diagnostic or prognostic biomarkers for human disease.

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Immunology and Infection

piggyBac Transposon System Modification of Primary Human T Cells
Sunandan Saha 1,2, Yozo Nakazawa 3, Leslie E. Huye 4,5, Joseph E. Doherty 2,6, Daniel L. Galvan 2, Cliona M. Rooney 4,5,7, Matthew H. Wilson 1,2,4,8
1Program in Translational Biology and Molecular Medicine, Baylor College of Medicine , 2Department of Medicine, Division of Nephrology, Baylor College of Medicine , 3Department of Immunology and Pathology, Shinshu University School of Medicine, 4Center for Cell and Gene Therapy, Baylor College of Medicine , 5Department of Pediatrics, Baylor College of Medicine , 6Program in Cell and Molecular Biology, Baylor College of Medicine , 7Department of Molecular Virology and Microbiology, Baylor College of Medicine , 8Michael E. DeBakey VA Medical Center

We describe a method to genetically modify primary human T cells with a transgene using the non-viral piggyBac transposon system. T cells modified to using the piggyBac transposon system exhibit stable transgene expression.

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Medicine

Neo-Islet Formation in Liver of Diabetic Mice by Helper-dependent Adenoviral Vector-Mediated Gene Transfer
Rongying Li 1,2, Kazuhiro Oka 1,2,3, Vijay Yechoor 1,2,3
1Department of Medicine, Baylor College of Medicine , 2Division of Diabetes, Endocrinology & Metabolism, Diabetes & Endocrinology Research Center, Baylor College of Medicine , 3Department of Molecular & Cellular Biology, Baylor College of Medicine

We describe hepatic neo-islet formation in STZ (streptozotocin)-induced diabetic mice by gene transfer of Neurogenin3 (Ngn3) and Betacellulin (Btc) using helper-dependent adenoviral vector (HDAd) and the reversal of hyperglycemia. Our method takes advantages of helper-dependent adenoviral vectors with their highly efficient in vivo transduction and the long lasting gene expression.

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Medicine

Assessing Phagocytic Clearance of Cell Death in Experimental Stroke by Ligatable Fluorescent Probes
Candace L. Minchew 1,2, Vladimir V. Didenko 1,2
1Baylor College of Medicine, 2Michael E. DeBakey Veterans Affairs Medical Center

We present a new fluorescence technique for selective in situ labeling of active phagocytic cells, which clear off cell corpses in stroke. The approach is important for assessing brain reaction to ischemia because only a small proportion of phagocytes present in ischemic brain participate in clearance of cell death.

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Biology

Hydrodynamic Renal Pelvis Injection for Non-viral Expression of Proteins in the Kidney
Lauren E. Woodard 1,2,3, Richard C. Welch 2, Felisha M. Williams 2, Wentian Luo 2, Jizhong Cheng 3, Matthew H. Wilson 1,2,3
1Department of Veterans Affairs, Tennessee Valley Healthcare System, 2Departments of Medicine and Pharmacology, Vanderbilt University Medical Center, 3Department of Medicine, Baylor University College of Medicine

This protocol describes a method to inject plasmid DNA into the mouse kidney via the renal pelvis to produce transgene expression specifically in the kidney.

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