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Rapid analysis of T-cell selection in vivo using T cell–receptor retrogenic mice

Nature Methods volume 3, pages 191–197 (2006)Cite this article

An Erratum to this article was published on 01 April 2006

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Abstract

Although T-cell receptor (TCR) transgenic as well as knockout and knockin mice have had a large impact on our understanding of T-cell development, signal transduction and function, the need to cross these mice delays experiments considerably. Here we provide a methodology for the rapid expression of TCRs in mice using 2A peptide–linked multicistronic retroviral vectors to transduce stem cells of any background before adoptive transfer into RAG-1−/− mice. For simplicity, we refer to these as retrogenic mice. We demonstrate that these retrogenic mice are comparable to transgenic mice expressing three commonly used TCRs (OT-I, OT-II and AND). We also show that retrogenic mice expressing male antigen–specific TCRs (HY, MataHari and Marilyn) facilitated the analysis of positive and negative selection in female and male mice, respectively. We examined various tolerance mechanisms in epitope-coupled TCR retrogenic mice. This powerful resource could expedite the identification of proteins involved in T-cell development and function.

* Note: In the version of this article initially published, the name of one of the receptors mentioned in the abstract was incorrectly stated as OY-II instead of OT-II. The correct sentence is: “We demonstrate that these retrogenic mice are comparable to transgenic mice expressing three commonly used TCRs (OT-I, OT-II, and AND). “ This error has been corrected in the HTML and PDF versions of the article.

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Figure 1: Comparison of TCR Rg and Tg mice.
Figure 2: Comparison of TCR Rg and Tg mice.
Figure 3: Analysis of tolerance induction using male HY antigen–specific Rg mice.
Figure 4: Analysis of tolerance induction using male HY antigen–specific Rg mice.
Figure 5: Analysis of CD4+ T-cell selection using H-2Eα–coupled TEa TCR Rg mice.
Figure 6: Analysis of CD8+ T-cell selection using peptide-coupled OT-I Rg mice.

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  • 01 March 2006

    In the version of this article initially published, the name of one of the receptors mentioned in the abstract was incorrectly stated as OY-II instead of OT-II. The correct sentence is: “We demonstrate that these retrogenic mice are comparable to transgenic mice expressing three commonly used TCRs (OT-I, OT-II, and AND). “ This error has been corrected in the HTML and PDF versions of the article.

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Acknowledgements

We are indebted to P. Matzinger, A. Rudensky, F. Carbone, M. Rubenstein, E. Parganas, S. Gingras and J. Ihle for reagents and information. We are grateful to S. Dilioglou and E. Vincent for technical assistance, K. Forbes for mouse colony management, R. Cross, J. Hoffrage and J. Smith for FACS analysis and sorting, S. Rowe for cytokine analysis, staff of the Flow Cytometry Shared Resource facility for MACS and the staff in the Hartwell Center for oligonucleotide synthesis and DNA sequencing. This work was supported by the NIH (AI–52199), a Cancer Center Support CORE grant (CA–21765) and the American Lebanese Syrian Associated Charities (ALSAC) (all to D.A.A.V.).

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Authors and Affiliations

  1. Department of Immunology, St. Jude Children's Research Hospital, 332 N. Lauderdale, Memphis, 38105, Tennessee, USA

    Jeff Holst, Kate M Vignali, Amanda R Burton & Dario A A Vignali

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  1. Jeff Holst
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  2. Kate M Vignali
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  3. Amanda R Burton
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  4. Dario A A Vignali
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Correspondence to Dario A A Vignali.

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Supplementary information

Supplementary Fig. 1

Comparison between transgenic and retrogenic mice. (PDF 135 kb)

Supplementary Fig. 2

Characterization of AND retrogenic mice. (PDF 78 kb)

Supplementary Fig. 3

Characterization of TEa retrogenic mice. (PDF 121 kb)

Supplementary Fig. 4

Characterization of male-specific TCR retrogenic mice. (PDF 120 kb)

Supplementary Fig. 5

Characterization of epitope-coupled OT-I retrogenic mice. (PDF 160 kb)

Supplementary Table 1

Description of the TCR constructs used in this study. (PDF 76 kb)

Supplementary Methods (PDF 33 kb)

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Holst, J., Vignali, K., Burton, A. et al. Rapid analysis of T-cell selection in vivo using T cell–receptor retrogenic mice. Nat Methods 3, 191–197 (2006). https://doi.org/10.1038/nmeth858

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