NDLI: MHC Class II Tetramers Made from Isolated Recombinant α and β Chains Refolded with Affinity-Tagged Peptides

Content Provider	PubMed Central
Author	Peter, Braendstrup Justesen, Sune Østerbye, Thomas Nielsen, Lise Lotte Bruun Mallone, Roberto Lars, Vindeløv Stryhn, Anette Buus, Søren
Editor	Dieli, Francesco
Copyright Year	2013
Abstract	Targeting CD4+ T cells through their unique antigen-specific, MHC class II-restricted T cell receptor makes MHC class II tetramers an attractive strategy to identify, validate and manipulate these cells at the single cell level. Currently, generating class II tetramers is a specialized undertaking effectively limiting their use and emphasizing the need for improved methods of production. Using class II chains expressed individually in E. coli as versatile recombinant reagents, we have previously generated peptide-MHC class II monomers, but failed to generate functional class II tetramers. Adding a monomer purification principle based upon affinity-tagged peptides, we here provide a robust method to produce class II tetramers and demonstrate staining of antigen-specific CD4+ T cells. We also provide evidence that both MHC class II and T cell receptor molecules largely accept affinity-tagged peptides. As a general approach to class II tetramer generation, this method should support rational CD4+ T cell epitope discovery as well as enable specific monitoring and manipulation of CD4+ T cell responses.
Related Links	http://dx.doi.org/10.1371/journal.pone.0073648
Starting Page	73648
File Format	PDF
ISSN	19326203
e-ISSN	19326203
Journal	PLoS ONE
Issue Number	9
Volume Number	8
Language	English
Publisher	Public Library of Science
Publisher Date	2013-09-01
Access Restriction	Open
Rights Holder	Public Library of Science
Subject Keyword	Biochemistry, Genetics and Molecular Biology(all) Agricultural and Biological Sciences(all) Medicine(all) Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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Correction: MHC Class II Tetramers Made from Isolated Recombinant α and β Chains Refolded with Affinity-Tagged Peptides

Correction: MHC Class II Tetramers Made from Isolated Recombinant α and β Chains Refolded with Affinity-Tagged Peptides

Structural Properties of MHC Class II Ligands, Implications for the Prediction of MHC Class II Epitopes

Anchor Side Chains of Short Peptide Fragments Trigger Ligand-Exchange of Class II MHC Molecules

One-Pot, Mix-and-Read Peptide-MHC Tetramers

Preserved MHC Class II Antigen Processing in Monocytes from HIV-Infected Individuals

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MHC Class II Tetramers Made from Isolated Recombinant α and β Chains Refolded with Affinity-Tagged Peptides

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Correction: MHC Class II Tetramers Made from Isolated Recombinant α and β Chains Refolded with Affinity-Tagged Peptides

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MHC Class II Tetramers Made from Isolated Recombinant α and β Chains Refolded with Affinity-Tagged Peptides