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Immunology and Infection

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JoVE Journal Immunology and Infection
Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques
 

Single-cell Quantitation of mRNA and Surface Protein Expression in Simian Immunodeficiency Virus-infected CD4+ T Cells Isolated from Rhesus macaques

Article DOI: 10.3791/57776-v • 13:13 min • September 25th, 2018
September 25th, 2018 •
1, 1, 1, 2, 1
1US Military HIV Research Program, Henry M. Jackson Foundation, Walter Reed Army Institute of Research, 2Vaccine Research Center, NIAID, NIH

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Described is a methodology to quantitate the expression of 96 genes and 18 surface proteins by single cells ex vivo, allowing for the identification of differentially expressed genes and proteins in virus-infected cells relative to uninfected cells. We apply the approach to study SIV-infected CD4+ T cells isolated from rhesus macaques.

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Single-cell Quantitation MRNA Surface Protein Expression Simian Immunodeficiency Virus CD4+ T Cells Rhesus Macaques Host-pathogen Interactions Productively Infected Cells Host Cofactors Therapeutic Interventions Quantitative Measures Protein And MRNA Commercially Available Reagents Cell-sorting Procedure Flow Cytometry Core Pre-PCR Molecular Biology Workstation Gene Expression Assays RNase DNase Free Tube Forward And Reverse Primers DNA Suspension Buffer 96 Well PCR Plate Surface Staining Viable Cells Compensation Samples Fluorescent Antibody Cocktail
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