Invasion of Ureaplasma diversum in Hep-2 cells

Marques, Lucas Miranda; Ueno, Priscilla M; Buzinhani, Melissa; Cortez, Beatriz Araujo; Neto, R. L.; Yamaguti, Maurício; Oliveira, Renata Cristina Gobbi de; Guimarães, Ana Márcia Sá; Monezi, Telma Alves; Braga, Antônio Carlos Ricardo; Machado-Santelli, Gláucia Maria; Timenetsky, Jorge

doi:10.1186/1471-2180-10-83

Cited by 22 publications

(47 citation statements)

References 26 publications

(35 reference statements)

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“…It is important to note that the assay used by DeSilva measures combined activity of PLC and phospholipase D (PLD) because both cleavage products are in the soluble fraction and the radioactively labeled hydrogen would be found in both cleavage products [39]. PLC activity has been reported in Ureaplasma diversum cells as well, and has been suggested to play a role in ureaplasma invasion in mammalian cells [40]. However, the detection method used the artificial substrate p-nitrophenylphosphorylcholine (p-NPPC), which can be hydrolyzed by several other enzymes that can hydrolyze phosphate esters, including PLD [41].…”

Section: Resultsmentioning

confidence: 99%

Comparative genome analysis of 19 Ureaplasma urealyticum and Ureaplasma parvumstrains

Paralanov

Duffy

et al. 2012

BMC Microbiol

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BackgroundUreaplasma urealyticum (UUR) and Ureaplasma parvum (UPA) are sexually transmitted bacteria among humans implicated in a variety of disease states including but not limited to: nongonococcal urethritis, infertility, adverse pregnancy outcomes, chorioamnionitis, and bronchopulmonary dysplasia in neonates. There are 10 distinct serotypes of UUR and 4 of UPA. Efforts to determine whether difference in pathogenic potential exists at the ureaplasma serovar level have been hampered by limitations of antibody-based typing methods, multiple cross-reactions and poor discriminating capacity in clinical samples containing two or more serovars.ResultsWe determined the genome sequences of the American Type Culture Collection (ATCC) type strains of all UUR and UPA serovars as well as four clinical isolates of UUR for which we were not able to determine serovar designation. UPA serovars had 0.75−0.78 Mbp genomes and UUR serovars were 0.84−0.95 Mbp. The original classification of ureaplasma isolates into distinct serovars was largely based on differences in the major ureaplasma surface antigen called the multiple banded antigen (MBA) and reactions of human and animal sera to the organisms. Whole genome analysis of the 14 serovars and the 4 clinical isolates showed the mba gene was part of a large superfamily, which is a phase variable gene system, and that some serovars have identical sets of mba genes. Most of the differences among serovars are hypothetical genes, and in general the two species and 14 serovars are extremely similar at the genome level.ConclusionsComparative genome analysis suggests UUR is more capable of acquiring genes horizontally, which may contribute to its greater virulence for some conditions. The overwhelming evidence of extensive horizontal gene transfer among these organisms from our previous studies combined with our comparative analysis indicates that ureaplasmas exist as quasi-species rather than as stable serovars in their native environment. Therefore, differential pathogenicity and clinical outcome of a ureaplasmal infection is most likely not on the serovar level, but rather may be due to the presence or absence of potential pathogenicity factors in an individual ureaplasma clinical isolate and/or patient to patient differences in terms of autoimmunity and microbiome.

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Section: Resultsmentioning

confidence: 99%

Comparative genome analysis of 19 Ureaplasma urealyticum and Ureaplasma parvumstrains

Paralanov

Duffy

et al. 2012

BMC Microbiol

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“…The pellets were suspended by washing twice with PBS and incubated with carbocyanine dye solution (Vybrant TM DiI cell‐labeling solution DiI; Molecular Probes, Eugene, OR, USA). Five microliters of Vibrant DiI (dilution, 1:200) was added to U. parvum cells in 1 ml of PBS and incubated for 40 min at 37°C (Marques et al., ). The labeled U. parvum cells were separated by centrifugation for 15 min at 15,000 × g at 20°C, washed twice with PBS, gently suspended in 2% FBS in DMEM cell culture medium.…”

Section: Methodsmentioning

confidence: 99%

Intracellular fate of Ureaplasma parvum entrapped by host cellular autophagy

et al. 2017

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Genital mycoplasmas, including Ureaplasma spp., are among the smallest human pathogenic bacteria and are associated with preterm birth. Electron microscopic observation of U. parvum showed that these prokaryotes have a regular, spherical shape with a mean diameter of 146 nm. U. parvum was internalized into HeLa cells by clathrin‐mediated endocytosis and survived for at least 14 days around the perinuclear region. Intracellular U. parvum reached endosomes in HeLa cells labeled with EEA1, Rab7, and LAMP‐1 within 1 to 3 hr. After 3 hr of infection, U. parvum induced the cytosolic accumulation of galectin‐3 and was subsequently entrapped by the autophagy marker LC3. However, when using atg7 −/− MEF cells, autophagy was inadequate for the complete elimination of U. parvum in HeLa cells. U. parvum also colocalized with the recycling endosome marker Rab11. Furthermore, the exosomes purified from infected HeLa cell culture medium included U. parvum. In these purified exosomes ureaplasma lipoprotein multiple banded antigen, host cellular annexin A2, CD9, and CD63 were detected. This research has successfully shown that Ureaplasma spp. utilize the host cellular membrane compartments possibly to evade the host immune system.

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“…However, there was not observed positive samples in vaginal lavages. U. diversum is capable of adhesion to tissue and it has been reported the rapid invasion of Hep-2 cell being located in the perinuclear region of the cell (Marques et al 2010). Thus, the absence of DNA found could be associated to the accession to the tissue or an intracell infection in the endometrium.…”

Section: Discussionmentioning

confidence: 99%

Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice

Silva

Ferreira

Oliveira

et al. 2016

An. Acad. Bras. Ciênc.

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Ureaplasma diversum is an opportunistic pathogen associated with uterine infl ammation, impaired embryo implantation, infertility, abortions, premature birth of calves and neonatal pneumonia in cattle. It has been suggested that the intra-uterine infection by Ureaplasma diversum can cause vascular changes that hinder the success of pregnancy. Thus, the aim of this study was to evaluate the changes of intrauterine site of A/J mice in estrus or proestrus phase inoculated with Ureaplasma diversum. The infection was monitored at 24, 48 and 72 hours by the PCR methodology to detect the Ureaplasma in the inoculation site and the profi le of circulating blood cells. Morphological changes, intensity of infl ammation and the production of cytokines were compared. The infected mice showed local infl ammation through the production of IFN-γ and TNF-α. Ureaplasma diversum infections in the reproductive tract of studied mice seemed to be associated with the production of pro-infl ammatory cytokines in uterine parenchyma. The levels of TNF-α of infected mice were dependent on the bacterial load of inoculated Ureaplasma. Uterine experimental infections by Ureaplasma diversum have not been mentioned yet and herein we presented the fi rst report of an intrauterine infection model in mice.

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Invasion of Ureaplasma diversum in Hep-2 cells

Cited by 22 publications

References 26 publications

Comparative genome analysis of 19 Ureaplasma urealyticum and Ureaplasma parvumstrains

Comparative genome analysis of 19 Ureaplasma urealyticum and Ureaplasma parvumstrains

Intracellular fate of Ureaplasma parvum entrapped by host cellular autophagy

Intra-uterine experimental infection by Ureaplasma diversum induces TNF-α mediated womb inflammation in mice

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