{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,24]],"date-time":"2026-10-24T01:48:37Z","timestamp":1792806517614,"version":"4.5.0"},"reference-count":31,"publisher":"National Academy of Sciences","issue":"21","content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2003,10,14]]},"abstract":"<jats:p>\n                    Tuberculosis remains a leading cause of death worldwide, despite the availability of effective chemotherapy and a vaccine. Bacillus Calmette\u2013Gu\u00e9rin (BCG), the tuberculosis vaccine, is an attenuated mutant of\n                    <jats:italic>Mycobacterium bovis<\/jats:italic>\n                    that was isolated after serial subcultures, yet the functional basis for this attenuation has never been elucidated. A single region (\n                    <jats:italic>RD1<\/jats:italic>\n                    ), which is absent in all BCG substrains, was deleted from virulent\n                    <jats:italic>M. bovis<\/jats:italic>\n                    and\n                    <jats:italic>Mycobacterium tuberculosis<\/jats:italic>\n                    strains, and the resulting \u0394\n                    <jats:italic>RD1<\/jats:italic>\n                    mutants were significantly attenuated for virulence in both immunocompromised and immunocompetent mice. The\n                    <jats:italic>M. tuberculosis<\/jats:italic>\n                    \u0394\n                    <jats:italic>RD1<\/jats:italic>\n                    mutants were also shown to protect mice against aerosol challenge, in a similar manner to BCG. Interestingly, the \u0394\n                    <jats:italic>RD1<\/jats:italic>\n                    mutants failed to cause cytolysis of pneumocytes, a phenotype that had been previously used to distinguish virulent\n                    <jats:italic>M. tuberculosis<\/jats:italic>\n                    from BCG. A specific transposon mutation, which disrupts the\n                    <jats:italic>Rv3874 Rv3875<\/jats:italic>\n                    (\n                    <jats:italic>cfp-10 esat-6<\/jats:italic>\n                    ) operon of\n                    <jats:italic>RD1<\/jats:italic>\n                    , also caused loss of the cytolytic phenotype in both pneumocytes and macrophages. This mutation resulted in the attenuation of virulence in mice, as the result of reduced tissue invasiveness. Moreover, specific deletion of each transcriptional unit of\n                    <jats:italic>RD1<\/jats:italic>\n                    revealed that three independent transcriptional units are required for virulence, two of which are involved in the secretion of ESAT-6 (6-kDa early secretory antigenic target). We conclude that the primary attenuating mechanism of bacillus Calmette\u2013Gu\u00e9rin is the loss of cytolytic activity mediated by secreted ESAT-6, which results in reduced tissue invasiveness.\n                  <\/jats:p>","DOI":"10.1073\/pnas.1635213100","type":"journal-article","created":{"date-parts":[[2003,10,14]],"date-time":"2003-10-14T15:05:46Z","timestamp":1066143946000},"page":"12420-12425","update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":674,"title":["The primary mechanism of attenuation of bacillus Calmette\u2013Gu\u00e9rin is a loss of secreted lytic function required for invasion of lung interstitial tissue"],"prefix":"10.1073","volume":"100","author":[{"given":"Tsungda","family":"Hsu","sequence":"first","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Suzanne M.","family":"Hingley-Wilson","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Bing","family":"Chen","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mei","family":"Chen","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Annie Z.","family":"Dai","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paul M.","family":"Morin","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Carolyn B.","family":"Marks","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeevan","family":"Padiyar","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Celia","family":"Goulding","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mari","family":"Gingery","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David","family":"Eisenberg","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Robert G.","family":"Russell","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Steven C.","family":"Derrick","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; 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Harold","family":"King","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"William R.","family":"Jacobs","sequence":"additional","affiliation":[{"name":"Howard Hughes Medical Institute, Departments of Pathology and Microbiology and Immunology, and Analytic Imaging Facility, Albert Einstein College of Medicine, Bronx, NY 10461; UCLA\u2013DOE Institute of Genomics and Proteomics, University of California, Los Angeles, CA 90095-1570; Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892; and Division of Infectious Diseases, Department of Medicine, Emory School of Medicine, Atlanta, GA 30303"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2003,10,13]]},"reference":[{"key":"e_1_3_2_1_2","doi-asserted-by":"crossref","unstructured":"Bloom B. 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