Multiplexed combinatorial tetramer staining in a mouse model of virus infection
Multiplexed combinatorial tetramer staining in a mouse model of virus infection
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DOI：
10.1016/j.jim.2010.06.003
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发表时间：
2010-08-31
影响因子：
2.2
通讯作者：
Kedzierska, Katherine
中科院分区：
文献类型：
--
作者：
Cukalac, Tania;Valkenburg, Sophie A.;Kedzierska, Katherine
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 (D(b)NP(366)(+)CD8(+) and D(b)PA(224)(+)CD8(+)) and subdominant (K(b)PB1(703)(+)CD8(+), D(b)PB1-F2(62)(+)CD8(+) and K(b)NS2(114)(+)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. (C) 2010 Elsevier B.V. All rights reserved.
