title: Apoptosis in HEp-2 cells infected with Ureaplasma diversum.
abstract: BACKGROUND
  Bacterial pathogens have many strategies for infecting and persisting in host cells. Adhesion, invasion and intracellular life are important features in the biology of mollicutes. The intracellular location of Ureaplasma diversum may trigger disturbances in the host cell. This includes activation or inhibition of pro and anti-apoptotic factors, which facilitate the development of host damage. The aim of the present study was to associate U. diversum infection in HEp-2 cells and apoptosis induction. Cells were infected for 72hs with four U. diversum clinical isolates and an ATCC strain. The U. diversum invasion was analyzed by Confocal Laser Scanning Microscopy and gentamicin invasion assay. The apoptosis was evaluated using pro-apoptotic and anti-apoptotic gene expression, and FITC Annexin V/Dead Cell Apoptosis Kit.
  
  
  RESULTS
  The number of internalized ureaplasma in HEp-2 cells increased significantly throughout the infection. The flow cytometry analysis with fluorochromes to detect membrane depolarization and gene expression for caspase 2, 3 and 9 increased in infected cells after 24 hours. However, after 72 hours a considerable decrease of apoptotic cells was observed.
  
  
  CONCLUSIONS
  The data suggests that apoptosis may be initially induced by some isolates in association with HEp-2 cells, but over time, there was no evidence of apoptosis in the presence of ureaplasma and HEp-2 cells. The initial increase and then decrease in apoptosis could be related to bacterial pathogen-associated molecular pattern (PAMPS). Moreover, the isolates of U. diversum presented differences in the studied parameters for apoptosis. It was also observed that the amount of microorganisms was not proportional to the induction of apoptosis in HEp-2 cells.
authors.affiliation: Instituto Multidisciplinar em Saúde, Núcleo de Tecnologia em Saúde, Universidade Federal da Bahia, Vitória da Conquista, Brazil. aline.amorim@usp.br.
authors.forename: Aline Teixeira
authors.initials: AT
authors.lastname: Amorim
authors.affiliation: Instituto Multidisciplinar em Saúde, Núcleo de Tecnologia em Saúde, Universidade Federal da Bahia, Vitória da Conquista, Brazil. lucasm@usp.br.|Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, São Paulo, Brazil. lucasm@usp.br.
authors.forename: Lucas Miranda
authors.initials: LM
authors.lastname: Marques
authors.affiliation: Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, São Paulo, Brazil. angelita.gusmao@hotmail.com.
authors.forename: Angelita Maria Oliveira Gusmão
authors.initials: AM
authors.lastname: Santos
authors.affiliation: Instituto Multidisciplinar em Saúde, Núcleo de Tecnologia em Saúde, Universidade Federal da Bahia, Vitória da Conquista, Brazil. hellenbahia@hotmail.com.
authors.forename: Hellen Braga
authors.initials: HB
authors.lastname: Martins
authors.affiliation: Instituto Multidisciplinar em Saúde, Núcleo de Tecnologia em Saúde, Universidade Federal da Bahia, Vitória da Conquista, Brazil. maysabarbosa_06@hotmail.com.
authors.forename: Maysa Santos
authors.initials: MS
authors.lastname: Barbosa
authors.affiliation: Instituto Multidisciplinar em Saúde, Núcleo de Tecnologia em Saúde, Universidade Federal da Bahia, Vitória da Conquista, Brazil. izarezende90@hotmail.com.
authors.forename: Izadora Souza
authors.initials: IS
authors.lastname: Rezende
authors.affiliation: Instituto Multidisciplinar em Saúde, Núcleo de Tecnologia em Saúde, Universidade Federal da Bahia, Vitória da Conquista, Brazil. ewerton_etu@hotmail.com.
authors.forename: Ewerton Ferraz
authors.initials: EF
authors.lastname: Andrade
authors.affiliation: Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, São Paulo, Brazil. guilhermebcampos@hotmail.com.
authors.forename: Guilherme Barreto
authors.initials: GB
authors.lastname: Campos
authors.affiliation: Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, São Paulo, Brazil. tassianl@hotmail.com.
authors.forename: Tássia Neves
authors.initials: TN
authors.lastname: Lobão
authors.affiliation: Instituto de Ciências Biomédicas, Departamento de Biologia Celular e do Desenvolvimento, Universidade de São Paulo, São Paulo, Brazil. biacortez@yahoo.com.br.
authors.forename: Beatriz Araujo
authors.initials: BA
authors.lastname: Cortez
authors.affiliation: Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, São Paulo, Brazil. maamo@usp.br.
authors.forename: Telma Alvez
authors.initials: TA
authors.lastname: Monezi
authors.affiliation: Instituto de Ciências Biomédicas, Departamento de Biologia Celular e do Desenvolvimento, Universidade de São Paulo, São Paulo, Brazil. glaucia.santelli@gmail.com.
authors.forename: Glaucia Maria
authors.initials: GM
authors.lastname: Machado-Santelli
authors.affiliation: Instituto de Ciências Biomédicas, Departamento de Microbiologia, Universidade de São Paulo, São Paulo, Brazil. joti@usp.br.
authors.forename: Jorge
authors.initials: J
authors.lastname: Timenetsky
mesh_terms: D008841:Actin Cytoskeleton / Q000648:ultrastructure; D017209:Apoptosis / Q000502:physiology*; D001422:Bacterial Adhesion; D053143:Caspase 2 / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D053453:Caspase 9 / Q000378:metabolism; D002470:Cell Survival; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression; D005839:Gentamicins / Q000494:pharmacology; D006367:HeLa Cells / Q000382:microbiology; D006801:Humans; D018613:Microscopy, Confocal; D000069452:Pathogen-Associated Molecular Pattern Molecules / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D018709:Statistics, Nonparametric; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014509:Ureaplasma / Q000187:drug effects / Q000472:pathogenicity*; D016869:Ureaplasma Infections / Q000503:physiopathology*
chemical_list: D005839:Gentamicins; D000069452:Pathogen-Associated Molecular Pattern Molecules; D014409:Tumor Necrosis Factor-alpha; D053143:Caspase 2; D053148:Caspase 3; D053453:Caspase 9