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B6.Rag2-deficient mice [Y. Shinkai et al., Cell 68, 855 (1992)]; B6.Thy1.1 and Thy1.2 Rag2-deficient mice bearing a TCRαβ Tg receptor specific for the HY male antigen [P. Kisielow, H. Bluthman, U. D. Staerz, M. Steinmetz, H. von Boehmer, Nature 333, 742 (1988)] were from the Centre Developement Techniques Avancées, Orleans, France.
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B6.Rag2-deficient mice [Y. Shinkai et al., Cell 68, 855 (1992)]; B6.Thy1.1 and Thy1.2 Rag2-deficient mice bearing a TCRαβ Tg receptor specific for the HY male antigen [P. Kisielow, H. Bluthman, U. D. Staerz, M. Steinmetz, H. von Boehmer, Nature 333, 742 (1988)] were from the Centre Developement Techniques Avancées, Orleans, France.
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1842280612
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note
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Host mice were irradiated at 600 rayleigh to spare BM precursors, but few peripheral cells were left. Peripheral T cell reconstitution requires previous recolonization of the thymus by host BM precursors, and host T cells start to emerge 1 month after irradiation. Irradiation may prevent the rejection of donor cells. It is also fundamental to allow quantification of T cell decays after adoptive transfer, because donor cells must be initially present at relatively high frequencies, requiring previous depletion of the host's lymphoid organs. In contrast to T cell decays, T cell survival or expansion can be quantified after adoptive transfer into intact mice.
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19
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26544476390
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in preparation
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2M-deficient mice [B. H. Koller, P. Marrack, J. W. Kapler, O. Smithies, Science 248, 1227 (1990)] were bred at the Pasteur Institute.
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Pérarnau, B.1
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20
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0030971268
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2M-deficient mice [B. H. Koller, P. Marrack, J. W. Kapler, O. Smithies, Science 248, 1227 (1990)] were bred at the Pasteur Institute.
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Pascolo, S.1
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2M-deficient mice [B. H. Koller, P. Marrack, J. W. Kapler, O. Smithies, Science 248, 1227 (1990)] were bred at the Pasteur Institute.
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Koller, B.H.1
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24
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1842352657
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unpublished data
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+ mice [B. Rocha, unpublished data].
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Rocha, B.1
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25
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1842386407
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note
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5 lymphocytes. In MHC-deficient hosts in which donor cells virtually disappear, we sorted T cells from all LN cell suspensions and half the spleen, to calculate their frequency.
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27
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1842324625
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note
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Bromodeoxyuridine (Sigma, 1 mg/mouse) was administered intraperitoneally twice a day for 3 days. Cell populations identified by surface staining were sorted in a FACS-Vintage (Becton Dickinson). Labeling with fluorescein isothiocyanate (FITC)-anti-BrdU mAb (Becton Dickinson) was done as described (10).
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29
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0030249953
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+ T cells, reported to survive for 6 months in mice lacking class II [S. Takeda, H.-R. Rodewald, H. Arakawa, H. Bluethman, T. Shimizu, Immunity 5, 217 (1996)]. However, fetal and adult T cell populations have such intrinsic differences in kinetic behavior [reviewed in (36)] and signal transduction mechanisms that the behavior of fetal cells may not be identical to those of adult T cells.
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Takeda, S.1
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Shimizu, T.5
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30
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1842392273
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unpublished data
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We used this system to immunize Tg cells rather than directly stimulating Tg female mice with male cells to generate homogeneous populations of activated Tg cells. In fact, when female Tg mice are immunized with male cells, a substantial fraction of Tg lymphocytes is never activated by male antigen (B. Rocha, unpublished data).
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Rocha, B.1
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31
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0029094319
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+, and antigen-presenting MHC class II-positive cells by magnetic sorting with coated Dynabeads (Dynal, Byosis, France). For surface staining, we used the following mAbs: biotin-labeled T3.70 (anti-TCRVα Tg) (15) (visualized with streptavidin TRICOLOR (Caltag, San Francisco, CA) or streptavidin-allophycocyanin (APC, Molecular Probes, Cambridge, UK); phycoerythrin-labeled mAbs to CD8, B220, and CD44 (Pharmingen, San Diego, CA); Red 16-labeled mAbs to CD8 (Gibco-BRL, UK); and FITC-labeled anti-Ly5.1 (A1042.1) and anti-Thy1.1 (19XE5) [M. W. Lostrom et al., Virology 98, 336 (1979)]. Cell populations were analyzed, and sorted in a FACS-Vintage (Becton Dickinson).
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Malissen, M.1
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32
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0018567955
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+, and antigen-presenting MHC class II-positive cells by magnetic sorting with coated Dynabeads (Dynal, Byosis, France). For surface staining, we used the following mAbs: biotin-labeled T3.70 (anti-TCRVα Tg) (15) (visualized with streptavidin TRICOLOR (Caltag, San Francisco, CA) or streptavidin-allophycocyanin (APC, Molecular Probes, Cambridge, UK); phycoerythrin-labeled mAbs to CD8, B220, and CD44 (Pharmingen, San Diego, CA); Red 16-labeled mAbs to CD8 (Gibco-BRL, UK); and FITC-labeled anti-Ly5.1 (A1042.1) and anti-Thy1.1 (19XE5) [M. W. Lostrom et al., Virology 98, 336 (1979)]. Cell populations were analyzed, and sorted in a FACS-Vintage (Becton Dickinson).
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Virology
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Lostrom, M.W.1
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34
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0024332461
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+ mice (70%). This difference is to be expected, because it has previously been demonstrated that the rate of cell expansion is dependent on "space" and is increased after adoptive transfer in mice containing few T cells [B. Rocha, N. Dautigny, P. Pereira, Eur. J. Immunol. 19, 905 (1989)].
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b also varies in different organs (B. Pérarnau et al., in preparation).
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46
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1842278655
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note
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+ cells persist in these hosts.
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47
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1842322744
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note
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b knockout mice were constructed.
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