title: Endocytosis‒Mediated Invasion and Pathogenicity of Streptococcus agalactiae in Rat Cardiomyocyte (H9C2).
abstract: Streptococcus agalactiae infection causes high mortality in cardiovascular disease (CVD) patients, especially in case of setting prosthetic valve during cardiac surgery. However, the pathogenesis mechanism of S. agalactiae associate with CVD has not been well studied. Here, we have demonstrated the pathogenicity of S. agalactiae in rat cardiomyocytes (H9C2). Interestingly, both live and dead cells of S. agalactiae were uptaken by H9C2 cells. To further dissect the process of S. agalactiae internalization, we chemically inhibited discrete parts of cellular uptake system in H9C2 cells using genistein, chlorpromazine, nocodazole and cytochalasin B. Chemical inhibition of microtubule and actin formation by nocodazole and cytochalasin B impaired S. agalactiae internalization into H9C2 cells. Consistently, reverse‒ transcription PCR (RT‒PCR) and quantitative real time‒PCR (RT-qPCR) analyses also detected higher levels of transcripts for cytoskeleton forming genes, Acta1 and Tubb5 in S. agalactiae‒infected H9C2 cells, suggesting the requirement of functional cytoskeleton in pathogenesis. Host survival assay demonstrated that S. agalactiae internalization induced cytotoxicity in H9C2 cells. S. agalactiae cells grown with benzyl penicillin reduced its ability to internalize and induce cytotoxicity in H9C2 cells, which could be attributed with the removal of surface lipoteichoic acid (LTA) from S. agalactiae. Further, the LTA extracted from S. agalactiae also exhibited dose‒dependent cytotoxicity in H9C2 cells. Taken together, our data suggest that S. agalactiae cells internalized H9C2 cells through energy‒dependent endocytic processes and the LTA of S. agalactiae play major role in host cell internalization and cytotoxicity induction.
authors.affiliation: Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625 021, India.
authors.forename: Sharma
authors.initials: S
authors.lastname: Pooja
authors.affiliation: Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625 021, India.
authors.forename: Muthuirulan
authors.initials: M
authors.lastname: Pushpanathan
authors.affiliation: Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625 021, India.
authors.forename: Paramasamy
authors.initials: P
authors.lastname: Gunasekaran
authors.affiliation: Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai, 625 021, India.
authors.forename: Jeyaprakash
authors.initials: J
authors.lastname: Rajendhran
mesh_terms: D000818:Animals; D002460:Cell Line; D004705:Endocytosis / Q000502:physiology*; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism; D032383:Myocytes, Cardiac / Q000378:metabolism / Q000382:microbiology*; D051381:Rats; D013290:Streptococcal Infections / Q000378:metabolism / Q000382:microbiology* / Q000473:pathology*; D013292:Streptococcus agalactiae / Q000472:pathogenicity*; D013682:Teichoic Acids / Q000378:metabolism; D014774:Virulence / Q000502:physiology
chemical_list: D008070:Lipopolysaccharides; D013682:Teichoic Acids; C009900:lipoteichoic acid