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A genome survey implicated sialidase and hyaluronidase, potential promoters of CD95-mediated eukaryotic cell death, as virulence factors of<jats:italic>M. alligatoris<\/jats:italic>. We used immunofluorescence imaging and flow cytometry to examine the effects of<jats:italic>M. alligatoris<\/jats:italic>infection in vitro on CD95 expression and apoptosis by alligator cardiac fibroblasts, a major cell type of a target organ of<jats:italic>M. alligatoris<\/jats:italic>infection in vivo. A uniform distribution of CD95 in primary cultured cardiac, skeletal muscle, and embryonic fibroblasts was demonstrated by using polyclonal antibodies against the N or C terminus of mouse or human CD95. Anti-CD95 antibodies reacted on Western blots of fibroblast lysates with a band with the predicted apparent molecular weight of CD95, but soluble CD95 was not detected in plasma from control or<jats:italic>M. alligatoris<\/jats:italic>-infected alligators. The proportion of CD95-gated cardiac fibroblasts increased threefold (<jats:italic>P<\/jats:italic>&lt; 0.01) 48 h after inoculation with<jats:italic>M. alligatoris<\/jats:italic>. Infection induced morphological changes in cardiac fibroblasts, including translocation of CD95 characteristic of apoptosis and an eightfold increase (<jats:italic>P<\/jats:italic>&lt; 0.16) in 5-bromo-2\u2032-deoxyuridine (BrdU) incorporation measured in a terminal deoxynucleotide transferase dUTP nick end-labeling apoptosis assay. The proportion of BrdU-gated controls activated with agonistic immunoglobulin M against human CD95 also increased threefold (<jats:italic>P<\/jats:italic>&lt; 0.03 for muscle). Heat-inactivated<jats:italic>M. alligatoris<\/jats:italic>and sterile<jats:italic>M. alligatoris<\/jats:italic>-conditioned culture supernatant had no effect. This is the first report of a CD95 homolog in the class Reptilia and establishes a new model that can be used to test the direct bacterial interaction with upstream components of the CD95 signal transduction pathway.<\/jats:p>","DOI":"10.1128\/cdli.12.12.1370-1377.2005","type":"journal-article","created":{"date-parts":[[2005,12,7]],"date-time":"2005-12-07T20:07:41Z","timestamp":1133986061000},"page":"1370-1377","update-policy":"https:\/\/doi.org\/10.1128\/asmj-crossmark-policy-page","source":"Crossref","is-referenced-by-count":6,"title":["<i>Mycoplasma alligatoris<\/i>Infection Promotes CD95 (FasR) Expression and Apoptosis of Primary Cardiac Fibroblasts"],"prefix":"10.1128","volume":"12","author":[{"given":"M. E.","family":"Hunt","sequence":"first","affiliation":[{"name":"Department of Pathobiology, College of Veterinary Medicine, University of Florida, Gainesville, Florida 32611-0880"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"D. 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