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MD Anderson Cancer Center - University of Texas

4 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Multicolor Flow Cytometry Analyses of Cellular Immune Response in Rhesus Macaques

Hong He¹, Amy N. Courtney¹, Eric Wieder², K. Jagannadha Sastry¹

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.

Immunology and Infection

Expansion, Purification, and Functional Assessment of Human Peripheral Blood NK Cells

Srinivas S. Somanchi¹, Vladimir V. Senyukov¹, Cecele J. Denman¹, Dean A. Lee¹

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.

Medicine

Assessment of Sarcoplasmic Reticulum Calcium Reserve and Intracellular Diastolic Calcium Removal in Isolated Ventricular Cardiomyocytes

Jing Gao^*1, Xiaolu Shi^*2, Hong He¹, Juhong Zhang¹, Ding Lin³, Guosheng Fu¹, Dongwu Lai¹

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.

Immunology and Infection

Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins

Meisam Naeimi Kararoudi¹, Hamid Dolatshad², Prashant Trikha¹, Syed-Rehan A. Hussain³, Ezgi Elmas¹, Jennifer A. Foltz¹, Jena E. Moseman¹, Aarohi Thakkar¹, Robin J. Nakkula¹, Margaret Lamb¹, Nitin Chakravarti¹, K. John McLaughlin³, Dean A. Lee¹

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).