{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,22]],"date-time":"2025-10-22T17:39:30Z","timestamp":1761154770764},"reference-count":29,"publisher":"Microbiology Society","issue":"5","content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2005,5,1]]},"abstract":"\n Actinobacillus actinomycetemcomitans<\/jats:italic> is a facultatively intracellular pathogen and the aetiological agent of localized aggressive periodontitis. Screening of the genome of A. actinomycetemcomitans<\/jats:italic> for in vivo<\/jats:italic>-induced antigen determinants previously demonstrated that the proteome of this organism differs in laboratory culture compared with conditions found during active infection. The aim of the present study was to determine whether the bacterial gene expression pattern inferred with in vivo<\/jats:italic>-induced antigen technology (IVIAT) in human infections was consistent with the gene expression pattern occurring upon epithelial cell association. To this end, a real-time PCR method was developed and used to quantify absolute and relative bacterial gene expression of A. actinomycetemcomitans<\/jats:italic> grown extra- and intracellularly in two human epithelial cell lines (HeLa and IHGK). The amount of template used in the assay was normalized using the total count of viable bacteria (c.f.u.) as a reference point and performed in duplicate in at least two independent experiments. Controls for this experiment included 16S rRNA and gapdh<\/jats:italic>. Transcription of all eight ORFs tested increased significantly (P<\/jats:italic> < 0.05) in HeLa and IHGK cells compared with bacteria grown extracellularly. The concurrence of gene expression patterns found in the two models suggests that these epithelial cells are valid in vitro<\/jats:italic> models of infection for the genes tested. IVIAT is an experimental platform that can be used as a validation tool to assess the reliability of animal and other models of infection and is applicable to most pathogens.<\/jats:p>","DOI":"10.1099\/jmm.0.45949-0","type":"journal-article","created":{"date-parts":[[2005,4,11]],"date-time":"2005-04-11T23:38:02Z","timestamp":1113262682000},"page":"497-504","source":"Crossref","is-referenced-by-count":18,"title":["Concurrence between the gene expression pattern of Actinobacillus actinomycetemcomitans in localized aggressive periodontitis and in human epithelial cells"],"prefix":"10.1099","volume":"54","author":[{"given":"Joseph","family":"Richardson","sequence":"first","affiliation":[{"name":"Center for Molecular Microbiology and Department of Oral Biology, 1600 SW Archer Road, University of Florida, Gainesville, FL 32610-0424, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Justin Corey","family":"Craighead","sequence":"additional","affiliation":[{"name":"Center for Molecular Microbiology and Department of Oral Biology, 1600 SW Archer Road, University of Florida, Gainesville, FL 32610-0424, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Sam Linsen","family":"Cao","sequence":"additional","affiliation":[{"name":"Center for Molecular Microbiology and Department of Oral Biology, 1600 SW Archer Road, University of Florida, Gainesville, FL 32610-0424, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Martin","family":"Handfield","sequence":"additional","affiliation":[{"name":"Center for Molecular Microbiology and Department of Oral Biology, 1600 SW Archer Road, University of Florida, Gainesville, FL 32610-0424, USA"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"345","reference":[{"key":"H1-15","doi-asserted-by":"crossref","first-page":"1125","DOI":"10.1046\/j.1365-2958.2003.03748.x","article-title":"& 12 other authors","volume":"50","author":"Asakawa","year":"2003","journal-title":"Mol Microbiol"},{"key":"H2-15","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1111\/j.1574-6968.2004.tb09683.x","article-title":"In vivo induced antigenic determinants of Actinobacillus actinomycetemcomitans","volume":"237","author":"Cao","year":"2004","journal-title":"FEMS Microbiol Lett"},{"key":"H3-15","doi-asserted-by":"crossref","first-page":"245","DOI":"10.1034\/j.1399-302X.2002.170407.x","article-title":"Kinetics of KB and HEp-2 cell responses to an invasive, cytolethal distending toxin-producing strain of Actinobacillus actinomycetemcomitans","volume":"17","author":"DiRienzo","year":"2002","journal-title":"Oral Microbiol Immunol"},{"key":"H4-15","doi-asserted-by":"crossref","first-page":"136","DOI":"10.1111\/j.1600-0757.1999.tb00161.x","article-title":"Virulence factors of Actinobacillus actinomycetemcomitans","volume":"20","author":"Fives-Taylor","year":"1999","journal-title":"Periodontol"},{"key":"H5-15","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1016\/S0966-842X(00)01775-3","article-title":"IVIAT: a novel method to identify microbial genes expressed specifically during human infections","volume":"8","author":"Handfield","year":"2000","journal-title":"Trends Microbiol"},{"key":"H6-15","first-page":"225","article-title":"In vivo expression of bacterial genes during human infections","volume":"71","author":"Handfield","year":"2003","journal-title":"Methods Mol Med"},{"key":"H7-15","doi-asserted-by":"crossref","DOI":"10.1111\/j.1462-5822.2005.00513.x","article-title":"Distinct transcriptional profiles characterize oral epithelium\u2013microbiota interactions","author":"Handfield","year":"2005","journal-title":"Cell Microbiol (in press)"},{"key":"H8-15","doi-asserted-by":"crossref","first-page":"928","DOI":"10.1128\/IAI.62.3.928-935.1994","article-title":"Characteristics of adherence of Actinobacillus actinomycetemcomitans to epithelial cells","volume":"62","author":"Meyer","year":"1994","journal-title":"Infect Immun"},{"key":"H9-15","doi-asserted-by":"crossref","first-page":"224","DOI":"10.1016\/S0966-842X(97)01055-X","article-title":"The role of Actinobacillus actinomycetemcomitans in the pathogenesis of periodontal disease","volume":"5","author":"Meyer","year":"1997","journal-title":"Trends Microbiol"},{"key":"H10-15","doi-asserted-by":"crossref","first-page":"2719","DOI":"10.1128\/IAI.59.8.2719-2726.1991","article-title":"Evidence for invasion of a human oral cell line by Actinobacillus actinomycetemcomitans","volume":"59","author":"Meyer","year":"1991","journal-title":"Infect Immun"},{"key":"H11-15","doi-asserted-by":"crossref","first-page":"2988","DOI":"10.1128\/IAI.64.8.2988-2997.1996","article-title":"Invasion of epithelial cells by Actinobacillus actinomycetemcomitans : a dynamic, multistep process","volume":"64","author":"Meyer","year":"1996","journal-title":"Infect Immun"},{"key":"H12-15","doi-asserted-by":"crossref","first-page":"6518","DOI":"10.1128\/IAI.67.12.6518-6525.1999","article-title":"Microtubules are associated with intracellular movement and spread of the periodontopathogen Actinobacillus actinomycetemcomitans","volume":"67","author":"Meyer","year":"1999","journal-title":"Infect Immun"},{"key":"H13-15","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1006\/excr.1996.0215","article-title":"HPV immortalization of human oral epithelial cells: a model for carcinogenesis","volume":"226","author":"Oda","year":"1996a","journal-title":"Exp Cell Res"},{"key":"H14-15","doi-asserted-by":"crossref","first-page":"2003","DOI":"10.1093\/carcin\/17.9.2003","article-title":"Chromosomal abnormalities in HPV-16-immortalized oral epithelial cells","volume":"17","author":"Oda","year":"1996b","journal-title":"Carcinogenesis"},{"key":"H15-15","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1902\/annals.1996.1.1.821","article-title":"Periodontal diseases: pathogenesis","volume":"1","author":"Offenbacher","year":"1996","journal-title":"Ann Periodontol"},{"key":"H16-15","doi-asserted-by":"crossref","first-page":"2700","DOI":"10.1128\/IAI.69.4.2700-2707.2001","article-title":"Intracellular Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in buccal epithelial cells collected from human subjects","volume":"69","author":"Rudney","year":"2001","journal-title":"Infect Immun"},{"key":"H17-15","doi-asserted-by":"crossref","first-page":"403","DOI":"10.1099\/jmm.0.05132-0","article-title":"Expression stability of six housekeeping genes : a proposal for resistance gene quantification studies of Pseudomonas aeruginosa by real-time quantitative RT-PCR","volume":"52","author":"Savli","year":"2003","journal-title":"J Med Microbiol"},{"key":"H18-15","doi-asserted-by":"crossref","first-page":"412","DOI":"10.1177\/00220345840630031101","article-title":"Black-pigmented Bacteroides species, Capnocytophaga species, and Actinobacillus actinomycetemcomitans in human periodontal disease: virulence factors in colonization, survival, and tissue destruction","volume":"63","author":"Slots","year":"1984","journal-title":"J Dent Res"},{"key":"H19-15","doi-asserted-by":"crossref","first-page":"239","DOI":"10.1016\/S0966-842X(98)01250-5","article-title":"What happens to bacterial pathogens in vivo ?","volume":"6","author":"Smith","year":"1998","journal-title":"Trends Microbiol"},{"key":"H20-15","doi-asserted-by":"crossref","first-page":"341","DOI":"10.1111\/j.1600-0757.1999.tb00165.x","article-title":"Ecological considerations in the treatment of Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis periodontal infections","volume":"20","author":"Socransky","year":"1999","journal-title":"Periodontol"},{"key":"H21-15","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1902\/annals.2002.7.1.38","article-title":"Genes of periodontopathogens expressed during human disease","volume":"7","author":"Song","year":"2002","journal-title":"Ann Periodontol"},{"key":"H22-15","doi-asserted-by":"crossref","first-page":"5598","DOI":"10.1128\/IAI.71.10.5598-5604.2003","article-title":"Augmentation of Actinobacillus actinomycetemcomitans invasion of human oral epithelial cells and up-regulation of interleukin-8 production by saliva CD14","volume":"71","author":"Takayama","year":"2003","journal-title":"Infect Immun"},{"key":"H23-15","doi-asserted-by":"crossref","first-page":"1086","DOI":"10.1016\/S0006-291X(03)00471-6","article-title":"Expression cloning of a periodontitis-associated apoptotic effector, cagE homologue, in Actinobacillus actinomycetemcomitans","volume":"303","author":"Teng","year":"2003","journal-title":"Biochem Biophys Res Commun"},{"key":"H24-15","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1006\/cyto.2001.0863","article-title":"Expression of IL-1\u03b2 and IL-8 by human gingival epithelial cells in response to Actinobacillus actinomycetemcomitans","volume":"14","author":"Uchida","year":"2001","journal-title":"Cytokine"},{"key":"H25-15","doi-asserted-by":"crossref","first-page":"7094","DOI":"10.1128\/JB.183.24.7094-7101.2001","article-title":"Quantification of expression of Staphylococcus epidermidis housekeeping genes with Taqman quantitative PCR during in vitro growth and under different conditions","volume":"183","author":"Vandecasteele","year":"2001","journal-title":"J Bacteriol"},{"key":"H26-15","doi-asserted-by":"crossref","first-page":"122","DOI":"10.1111\/j.1600-0757.1999.tb00160.x","article-title":"Actinobacillus actinomycetemcomitans and Porphyromonas gingivalis in nonoral infections","volume":"20","author":"van Winkelhoff","year":"1999","journal-title":"Periodontol"},{"key":"H27-15","doi-asserted-by":"crossref","first-page":"51","DOI":"10.1111\/j.1574-6968.2001.tb10779.x","article-title":"Quantification of the carbazole 1,9a-dioxygenase gene by real-time competitive PCR combined with co-extraction of internal standards","volume":"202","author":"Widada","year":"2001","journal-title":"FEMS Microbiol Lett"},{"key":"H28-15","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1111\/j.1600-051X.1985.tb01348.x","article-title":"Actinobacillus actinomycetemcomitans in human periodontal disease","volume":"12","author":"Zambon","year":"1985","journal-title":"J Clin Periodontol"},{"key":"H29-15","doi-asserted-by":"crossref","first-page":"38","DOI":"10.1128\/IAI.72.1.38-45.2004","article-title":"Long-term effect of heat shock protein 60 from Actinobacillus actinomycetemcomitans on epithelial cell viability and mitogen-activated protein kinases","volume":"72","author":"Zhang","year":"2004","journal-title":"Infect Immun"}],"container-title":["Journal of Medical Microbiology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.45949-0?crawler=true","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2020,9,23]],"date-time":"2020-09-23T03:17:42Z","timestamp":1600831062000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.microbiologyresearch.org\/content\/journal\/jmm\/10.1099\/jmm.0.45949-0"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2005,5,1]]},"references-count":29,"journal-issue":{"issue":"5","published-print":{"date-parts":[[2005,5,1]]}},"URL":"https:\/\/doi.org\/10.1099\/jmm.0.45949-0","relation":{},"ISSN":["0022-2615","1473-5644"],"issn-type":[{"value":"0022-2615","type":"print"},{"value":"1473-5644","type":"electronic"}],"subject":[],"published":{"date-parts":[[2005,5,1]]}}}