MHC Class II Tetramer Labeling of Human Primary CD4⁺ T Cells from HIV Infected Patients.

Content Provider	Europe PMC
Author	Galperin, Moran Benati, Daniela Claireaux, Mathieu Mukhopadhyay, Madhura Chakrabarti, Lisa A.
Copyright Year	2017
Description	Major Histocompatibility Complex (MHC) tetramers have been used for two decades to detect, isolate and characterize T cells specific for various pathogens and tumor antigens. In the context of Human Immunodeficiency Virus (HIV) infection, antigen-specific CD8+ T cells have been extensively studied ex vivo, as they can be readily detected by HIV peptide-loaded MHC class I tetramers. In contrast, the detection of HIV-specific CD4+ T cells has proven more challenging, due to the intrinsically lower clonal expansion rates of CD4+ T cells, and to the preferential depletion of HIV-specific CD4+ T cells in the course of HIV infection. In the following protocol, we describe a simple method that facilitates the identification of CD4+ T cells specific for an HIV-1 capsid epitope using peptide-loaded MHC class II tetramers. Tetramer labeled CD4+ T cells can be analyzed for their cell surface phenotype and/or FACS-sorted for further downstream applications. A key point for successful detection of specific CD4+ T cells ex vivo is the choice of a peptide/MHC II combination that results in high-affinity T Cell Receptor (TCR) binding ( Benati et al., 2016 ). A second key point for reliable detection of MHC II tetramer-positive cells is the systematic use of a control tetramer loaded with an irrelevant peptide, with the sample and control tubes being processed in identical conditions.
Abstract	Major Histocompatibility Complex (MHC) tetramers have been used for two decades to detect, isolate and characterize T cells specific for various pathogens and tumor antigens. In the context of Human Immunodeficiency Virus (HIV) infection, antigen-specific CD8+ T cells have been extensively studied ex vivo, as they can be readily detected by HIV peptide-loaded MHC class I tetramers. In contrast, the detection of HIV-specific CD4+ T cells has proven more challenging, due to the intrinsically lower clonal expansion rates of CD4+ T cells, and to the preferential depletion of HIV-specific CD4+ T cells in the course of HIV infection. In the following protocol, we describe a simple method that facilitates the identification of CD4+ T cells specific for an HIV-1 capsid epitope using peptide-loaded MHC class II tetramers. Tetramer labeled CD4+ T cells can be analyzed for their cell surface phenotype and/or FACS-sorted for further downstream applications. A key point for successful detection of specific CD4+ T cells ex vivo is the choice of a peptide/MHC II combination that results in high-affinity T Cell Receptor (TCR) binding (Benati et al., 2016). A second key point for reliable detection of MHC II tetramer-positive cells is the systematic use of a control tetramer loaded with an irrelevant peptide, with the sample and control tubes being processed in identical conditions.
Related Links	https://europepmc.org/backend/ptpmcrender.fcgi?accid=PMC8376602&blobtype=pdf
Volume Number	7
PubMed Central reference number	PMC8376602
Issue Number	6
PubMed reference number	34458496
Journal	Bio-protocol [Bio Protoc]
e-ISSN	23318325
DOI	10.21769/bioprotoc.2187
Language	English
Publisher	Bio-Protocol
Publisher Date	2017-03-20
Publisher Place	1075 Lorne Way, Sunnyvale, CA 94087, USA
Access Restriction	Open
Rights License	Copyright © 2017 The Authors; exclusive licensee Bio-protocol LLC.
Subject Keyword	Major histocompatibility complex class II Tetramer T cell receptor CD4+ T cell HIV
Content Type	Text
Resource Type	Article
Subject	Immunology and Microbiology Neuroscience Plant Science Biochemistry, Genetics and Molecular Biology