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- Publisher Website: 10.1016/j.jim.2010.06.003
- Scopus: eid_2-s2.0-77955657270
- PMID: 20558170
- WOS: WOS:000282147700018
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Article: Multiplexed combinatorial tetramer staining in a mouse model of virus infection
Title | Multiplexed combinatorial tetramer staining in a mouse model of virus infection |
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Authors | |
Keywords | Viral infection Mouse B6 model Cytotoxic T cells T cells PMHC tetramers |
Issue Date | 2010 |
Citation | Journal of Immunological Methods, 2010, v. 360, n. 1-2, p. 157-161 How to Cite? |
Abstract | Use of fluorescently labelled multimers, particularly tetramers of peptide and MHC class I glycoprotein (pMHC-I) complexes, is essential for the analysis of CD8+ T cell immunity in basic research and clinical settings. A recently described combinatorial approach using pMHC-I multimers coupled to a unique combination of distinct fluorochromes has facilitated the simultaneous screening of multiple T cell specificities within a single human blood sample. The present analysis establishes that this multiplexed tetramer staining protocol can also be applied in mouse models of a disease to detect multiple subdominant CD8+ T cell specificities in the presence of prominent immunodominant T cell sets at different stages of infection. We have established a modified protocol that concurrently identified influenza-specific CD8+ T cells at the acute and long-term memory phases of influenza virus infection in B6 mice. Highly dominant (DbNP366+CD8+ and DbPA224+CD8+) and subdominant (KbPB1703+CD8+, DbPB1-F262+CD8+ and KbNS2114+CD8+) T cell responses can be detected simultaneously at levels comparable to the conventional tetramer staining with this combinatorial approach. The technique proved particularly useful with aged mice, where we used 5-fold fewer animals, making the detection of multiple T cell specificities more cost-effective and less time-consuming. Overall, our study establishes that this comprehensive concurrent analysis of multiple T cell specificities is of value for analysing mouse models of disease, especially in situations where sample size and/or response magnitude is limiting. © 2010 Elsevier B.V. |
Persistent Identifier | http://hdl.handle.net/10722/241186 |
ISSN | 2021 Impact Factor: 2.287 2020 SCImago Journal Rankings: 0.870 |
ISI Accession Number ID |
DC Field | Value | Language |
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dc.contributor.author | Cukalac, Tania | - |
dc.contributor.author | Valkenburg, Sophie A. | - |
dc.contributor.author | La Gruta, Nicole L. | - |
dc.contributor.author | Turner, Stephen J. | - |
dc.contributor.author | Doherty, Peter C. | - |
dc.contributor.author | Kedzierska, Katherine | - |
dc.date.accessioned | 2017-05-26T03:37:03Z | - |
dc.date.available | 2017-05-26T03:37:03Z | - |
dc.date.issued | 2010 | - |
dc.identifier.citation | Journal of Immunological Methods, 2010, v. 360, n. 1-2, p. 157-161 | - |
dc.identifier.issn | 0022-1759 | - |
dc.identifier.uri | http://hdl.handle.net/10722/241186 | - |
dc.description.abstract | Use of fluorescently labelled multimers, particularly tetramers of peptide and MHC class I glycoprotein (pMHC-I) complexes, is essential for the analysis of CD8+ T cell immunity in basic research and clinical settings. A recently described combinatorial approach using pMHC-I multimers coupled to a unique combination of distinct fluorochromes has facilitated the simultaneous screening of multiple T cell specificities within a single human blood sample. The present analysis establishes that this multiplexed tetramer staining protocol can also be applied in mouse models of a disease to detect multiple subdominant CD8+ T cell specificities in the presence of prominent immunodominant T cell sets at different stages of infection. We have established a modified protocol that concurrently identified influenza-specific CD8+ T cells at the acute and long-term memory phases of influenza virus infection in B6 mice. Highly dominant (DbNP366+CD8+ and DbPA224+CD8+) and subdominant (KbPB1703+CD8+, DbPB1-F262+CD8+ and KbNS2114+CD8+) T cell responses can be detected simultaneously at levels comparable to the conventional tetramer staining with this combinatorial approach. The technique proved particularly useful with aged mice, where we used 5-fold fewer animals, making the detection of multiple T cell specificities more cost-effective and less time-consuming. Overall, our study establishes that this comprehensive concurrent analysis of multiple T cell specificities is of value for analysing mouse models of disease, especially in situations where sample size and/or response magnitude is limiting. © 2010 Elsevier B.V. | - |
dc.language | eng | - |
dc.relation.ispartof | Journal of Immunological Methods | - |
dc.subject | Viral infection | - |
dc.subject | Mouse B6 model | - |
dc.subject | Cytotoxic T cells | - |
dc.subject | T cells | - |
dc.subject | PMHC tetramers | - |
dc.title | Multiplexed combinatorial tetramer staining in a mouse model of virus infection | - |
dc.type | Article | - |
dc.description.nature | link_to_subscribed_fulltext | - |
dc.identifier.doi | 10.1016/j.jim.2010.06.003 | - |
dc.identifier.pmid | 20558170 | - |
dc.identifier.scopus | eid_2-s2.0-77955657270 | - |
dc.identifier.volume | 360 | - |
dc.identifier.issue | 1-2 | - |
dc.identifier.spage | 157 | - |
dc.identifier.epage | 161 | - |
dc.identifier.isi | WOS:000282147700018 | - |
dc.identifier.issnl | 0022-1759 | - |