A phosphoramidon-sensitive metalloprotease induces apoptosis of human endothelial cells by Group B Streptococcus
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ORIGINAL PAPER
A phosphoramidon-sensitive metalloprotease induces apoptosis of human endothelial cells by Group B Streptococcus Michelle Hanthequeste Bittencourt dos Santos • Andre´ia Ferreira Eduardo da Costa Beatriz Jandre Ferreira • Simone Lima Souza • Pamella da Silva Lannes • Gabriela Silva Santos • Ana Luiza Mattos-Guaraldi • Prescilla Emy Nagao
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Received: 5 June 2013 / Accepted: 6 September 2013 / Published online: 20 September 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract We explored Group B Streptococcus (GBS)-induced apoptosis in human umbilical vein endothelial cells (HUVEC) and the role of phosphoramidon, a zinc metalloprotease inhibitor, in this process. GBS 90186 strain (serotype V, a blood isolate) and concentrated supernatant (CS) were used to investigate the viability and morphological alterations in HUVEC by Trypan blue uptake, electrophoresis in 2 % agarose gel and scanning electron microscopy assays. Apoptosis before and after phosphoramidon-
M. H. B. dos Santos A. F. E. da Costa B. J. Ferreira P. da Silva Lannes G. S. Santos P. E. Nagao (&) Departamento de Biologia Celular, Instituto de Biologia Roberto Alcantara Gomes, Universidade do Estado do Rio de Janeiro, R. Sa˜o Francisco Xavier, 524 – PHLC, 5° andar, sala 501B, Maracana˜, Rio de Janeiro CEP 20.550-013, Brazil e-mail: [email protected] S. L. Souza Departamento de Patologia Geral e Laborato´rios, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil e-mail: [email protected] A. L. Mattos-Guaraldi Disciplina de Microbiologia e Imunologia, Faculdade de Cieˆncias Me´dicas, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil e-mail: [email protected]
treatment were verified by flow cytometry using annexin V-FITC labeling. Differences were considered significant when P \ 0.05 using unpaired Student’s t test. GBS and CS induced HUVEC death by apoptosis (76.5 and 32 %, respectively) with an increasing pro-apoptotic Bax expression and decreasing anti-apoptotic Bcl-2 expression. Caspase-3 was activated during GBS-induced endothelial apoptosis. Phosphoramidon reduced 89.3 and 100 % of GBS and CS cell death by apoptosis, respectively. Some GBS strains may induce cell death by apoptosis with involvement of metalloproteases and signaling through the intrinsic pathway of apoptosis, which may contribute to GBS survival during sepsis of adults and neonates. Keywords Group B Streptococcus Endothelial cells Metalloprotease Phosphoramidon Apoptosis
Introduction Group B Streptococcus (GBS; Streptococcus agalactiae) emerged in the 1960s as a major cause of neonatal morbidity and mortality in the United States and Europe. Three decades later, GBS arose as a pathogen responsible for various infections in non pregnant adults, particularly in elderly subjects with
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underlying conditions (Farley 1993). Primary bacteremia is the most serious clinical syndrome reported in adults and meningitis is associated with very high mortality rates (27–34 %) (Salloum et al. 2011). Disruption of
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