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MD Anderson Cancer Center - University of Texas

4 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques
Hong He 1, Amy N. Courtney 1, Eric Wieder 2, K. Jagannadha Sastry 1
1Department of Immunology, MD Anderson Cancer Center - University of Texas, 2Department of Medicine, University of Miami

We demonstrate the utility of multicolor flow cytometry for detailed phenotypic and functional characterization of total as well as memory subsets of CD4+ and CD8+ T cells in rhesus macaques, the ideal model for HIV/AIDS vaccine studies.

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Immunology and Infection

Expansion, Purification, and Functional Assessment of Human Peripheral Blood NK Cells
Srinivas S. Somanchi 1, Vladimir V. Senyukov 1, Cecele J. Denman 1, Dean A. Lee 1
1Division of Pediatrics, MD Anderson Cancer Center - University of Texas

Here we describe a method to efficiently expand and purify large numbers of human NK cells and assess their function.

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Medicine

Assessment of Sarcoplasmic Reticulum Calcium Reserve and Intracellular Diastolic Calcium Removal in Isolated Ventricular Cardiomyocytes
Jing Gao *1, Xiaolu Shi *2, Hong He 1, Juhong Zhang 1, Ding Lin 3, Guosheng Fu 1, Dongwu Lai 1
1Department of Cardiology, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, 2Experimental Research Center, China Academy of Chinese Medical Sciences, 3Department of Cardiology, The Third Hospital of Hangzhou City

Intracellular calcium recycling plays a critical role in regulation of systolic and diastolic function in cardiomyocytes. Here, we describe a protocol to evaluate sarcoplasmic reticulum Ca2+ reserve and diastolic calcium removal function in cardiomyocytes by a calcium imaging system.

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Immunology and Infection

Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins
Meisam Naeimi Kararoudi 1, Hamid Dolatshad 2, Prashant Trikha 1, Syed-Rehan A. Hussain 3, Ezgi Elmas 1, Jennifer A. Foltz 1, Jena E. Moseman 1, Aarohi Thakkar 1, Robin J. Nakkula 1, Margaret Lamb 1, Nitin Chakravarti 1, K. John McLaughlin 3, Dean A. Lee 1
1Center for Childhood Cancer and Blood Disease, Nationwide Children's Hospital, 2Nuffield Division of Clinical Laboratory Sciences, Radcliffe Department of Medicine, University of Oxford, 3Center for Clinical and Translational Research, Nationwide Children's Hospital Research Institute

Here, we present a protocol to genetically modify primary or expanded human natural killer (NK) cells using Cas9 Ribonucleoproteins (Cas9/RNPs). By using this protocol, we generated human NK cells deficient for transforming growth factor–b receptor 2 (TGFBR2) and hypoxanthine phosphoribosyltransferase 1 (HPRT1).

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