Information de reference pour ce titreAccession Number: | 00007529-200107130-00025.
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Author: | Dupuis, Stephanie 1,2; Dargemont, Catherine 3; Fieschi, Claire 1; Thomassin, Nicolas 4; Rosenzweig, Sergio 5; Harris, Jeff 6; Holland, Steven M. 5; Schreiber, Robert D. 7; Casanova, Jean-Laurent 1,8*
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Institution: | (1)Laboratoire de Genetique Humaine des Maladies Infectieuses, Universite de Paris Rene Descartes-INSERM UMR550, Faculte de Medecine Necker-Enfants Malades, 75015 Paris, France. (2)INSERM U429, Hopital Necker-Enfants Malades, 75015 Paris, France. (3)Institut Jacques Monod, CNRS-Universite Paris VI-Universite Paris VII-CNRS UMR7592, 75005 Paris, France. (4)Department de Pediatrie, Centre Hospitalier General, 58000 Nevers, France. (5)Laboratory of Host Defenses, National Institute of Allergy and Infectious Diseases, Bethesda, MD 20892-1886, USA. (6)Department of Pediatrics, University of California at San Francisco, San Francisco, CA 94117, USA. (7)Department of Pathology, Washington University, St. Louis, MO 63110, USA. (8)Unite d'Hematologie-Immunologie Pediatrique, Hopital Necker-Enfants Malades, 75015 Paris, France.
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Title: | Impairment of Mycobacterial But Not Viral Immunity by a Germline Human STAT1 Mutation.[Report]
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Source: | Science. Vaccines and Immunity. 293(5528):300-303, July 13, 2001.
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Abstract: | Interferons (IFN) [alpha]/[beta] and [gamma] induce the formation of two transcriptional activators: gamma-activating factor (GAF) and interferon-stimulated gamma factor 3 (ISGF3). We report a natural heterozygous germline STAT1 mutation associated with susceptibility to mycobacterial but not viral disease. This mutation causes a loss of GAF and ISGF3 activation but is dominant for one cellular phenotype and recessive for the other. It impairs the nuclear accumulation of GAF but not of ISGF3 in heterozygous cells stimulated by IFNs. Thus, the antimycobacterial, but not the antiviral, effects of human IFNs are principally mediated by GAF.
Copyright (C) 2001 by the American Association for the Advancement of Science
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21. The STAT1 cDNA was amplified with sense (5'-TCgACAgTCTTggCACCTAACgTgC-3') and antisense (5'-TgCTATCAACaggTTgCAgCg-3') primers. The STAT1 exon encoding L706 was amplified with sense (5'-TCggTTgATggAAagCgTA-3') and antisense (5'-CTCTTCTgTgTTCACTTAC-3') primers. Genomic DNA from various tissues (blood, hair roots, and buccal cells) and cell lines (EBV-B cells and SV40 fibroblasts) was amplified. The products were sequenced as previously described (10).
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23. Two days after electroporation with pcDNA3-WT-STAT-1 (provided by M. J. Holtzman), pcDNA3-L706S-STAT-1, or a mock vector, mouse STAT-1-deficient embryonic fibroblasts were stimulated with murine IFN-[gamma] (R&D) or IFN-[alpha] (Gibco BRL) for 30 min. Indirect immunofluorescence, with a mouse monoclonal antibody to human STAT-1 (N-Terminus; Transduction Laboratories) or a rabbit antibody specific to STAT-1-phosphotyrosine 701 (9171L; New England Biolabs) was performed as in (18). Double immunofluorescence was performed by incubation with a mouse antibody to human STAT-1 (N-Terminus; Transduction Laboratories) and a rabbit antibody to mouse STAT-2 Ab (provided by C. Schindler) for 30 min, followed by a tetramethyl rhodamine isothiocyanate-conjugated mouse antibody (Jackson) and FITC-conjugated rabbit antibody (Jackson) for 30 min as described by C. Park et al. [Nucleic Acids Res. 27, 4191 (1999)]. The cells were visualized by confocal microscopy (Zeiss).
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33. We thank L. Abel, S. Pellegrini, J. Puck, and J. C. Weill for helpful discussions and M. J. Holtzman, D. Frank, D. Recan, and C. Schindler for providing cells or reagents. J.-L.C. would like to thank A. Munnich for support and encouragement. This work was supported by grants from Action Incitative Concertee Blanche, Banque Nationale de Paris-Paribas, Fondation pour la Recherche Medicale, Fondation Schlumberger, Legs Poix, Programme National de Recherche Fondamentale en Microbiologie, Maladies Infectieuses et Parasitaires, and Institut Universitaire de France.
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Language: | English.
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Document Type: | Research: Reports.
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Journal Subset: | Life Sciences. Physical Science & Engineering.
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ISSN: | 0036-8075
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NLM Journal Code: | 0404511, uj7
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