POL Scientific / JBM / Volume 5 / Issue 3 / DOI: 10.14440/jbm.2018.246
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RNA isolation from Peyer's patch lymphocytes and mononuclear phagocytes to determine gene expression profiles using NanoString technology

Navjot Singh2 Heather C. Gallagher1,4 Renjie Song3 Jaskiran K. Dhinsa1,2 Gary R. Ostroff5 Magdia De Jesus1,4
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1 Division of Infectious Diseases, Wadsworth Center, New York State Department of Health, Albany NY, USA
2 Division of Molecular Genetics Wadsworth Center, New York State Department of Health, Albany NY, USA
3 Biochemistry and Immunology Core, Wadsworth Center, New York State Department of Health, Albany NY, USA
4 Department of Biomedical Sciences, University at Albany, School of Public Health Albany, Albany NY, USA
5 Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA
Published: 2 July 2018
© 2018 by the author. Licensee POL Scientific, USA. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )
Abstract

Sampling and immune surveillance within gut-associated lymphoid tissues (GALT) such as the intestinal Peyer’s patch (PP) occurs by an elegantly orchestrated effort that involves the epithelial barrier, B and T lymphocytes, and an extensive network of mononuclear phagocytes. Although we now understand more about the dynamics of antigen and microbial sampling within PPs, the gene expression changes that occur in individual cell subsets during sampling are not well characterized. This protocol describes the isolation of high-quality RNA from sorted PP, B and T-lymphocytes, and CD11c+ phagocytes for use with nCounter-nanostring technology. This method allows investigators to study gene expression changes within PPs in response to antigens, microbes, and oral vaccine delivery vehicles of interest that are sampled.

Keywords
B-lymphocytes
mononuclear phagocytes
Peyer
Nanostring technology
T-lymphocytes.
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Journal of Biological Methods, Electronic ISSN: 2326-9901 Print ISSN: TBA, Published by POL Scientific