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T cells were isolated and stained and analyzed as described by P. Marrack, J. Kappler, and T. Mitchell [J. Exp. Med. 189, 521 (1999)] with antibodies from PharMingen (San Diego, CA). Cells were labeled with CFSE (Molecular Probes, OR) by the method of S. A. Weston and C. R. Parish [J. Immunol. Methods 133, 87 (1990)]. Incorporation of BrdU into cellular DNA was measured as described by P. Carayon and A. Bord [J. Immunol. Methods 147, 225 (1992)] with anti-BrdU (Becton Dickinson, San Jose, CA). Mice were purchased from the Jackson Laboratory (Bar Harbor, ME) or from the National Institute of Aging mouse colony at Charles River Laboratories (Willington, MA). Mice were maintained in a pathogen-free environment in accordance with institutional guidelines.
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note
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The authors thank S. Nishikawa, M. Miyasaka, E. Shevach, T. R. Malek, A. Troutt, T. Mossman, and R. Coffman for monoclonal antibody-producing cell lines; PharMingen (San Diego, CA) for providing cell lines with the producers' permission; T. Potter for the OT1 mice; K. Christiansen for preparation of antibody-containing supernatants; W. Townend and S. Sobus for flow cytometry advice; and T. Mitchell and B. Schaefer for comments on the manuscript. This work was supported by NIH grants Al-17134, Al-18785, Al-22295, and CA-46934 and a fellowship for research abroad from the Japan Society for the Promotion of Science (1999-2001) (M.M.).
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