The influence of centrifugation and incubation temperatures on various veterinary and human chlamydial species

Onorini, Delia; Donati, Manuela; Marti, H. R.; Biondi, Roberta; Levi, Aurora; Nufer, Lisbeth; Prähauser, Barbara; Rigamonti, Sara; Vicari, Nadia; Borel, Nicole

doi:10.1016/j.vetmic.2019.04.012

Cited by 14 publications

(12 citation statements)

References 36 publications

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“…CtE was propagated as previously reported in LLC cells ( Marangoni et al., 2014 ), and crude stocks were suspended in a sucrose phosphate glutamate buffer (SPG; 218 mM sucrose (Sigma-Aldrich/MilliporeSigma); 3.76 mM KH 2 PO 4 (Sigma-Aldrich/MilliporeSigma), 7.1 mM K 2 HPO 4 (Sigma-Aldrich/MilliporeSigma), 5 mM GlutaMAX (Gibco, Thermo Fisher Scientific)) and stored at -80°C until use. Inclusion forming unit (IFU)/well determination was carried out in LLC and HeLa cells as previously described ( Leonard et al., 2015 ; Onorini et al., 2019 ). Equivalent infection of HeLa, as compared to LLC, resulted in similar stock IFU/mL determination.…”

Section: Methodsmentioning

confidence: 99%

“…At experimental endpoints, culture medium was removed from infected cells, cells were fixed with ice cold absolute methanol for 10 minutes and methanol was replaced with sterile PBS. Chlamydiae were immunostained as previously reported ( Onorini et al., 2019 ) with some modification of antibody concentrations: 1:400-diluted Chlamydiaceae family-specific mouse monoclonal chlamydial anti-lipopolysaccharide (LPS) antibody (Clone ACI-P, Progen, Heidelberg, Germany) and 1:750-diluted Alexa Fluor 488-conjugated secondary goat anti-mouse antibody (Alexa 488; Life Technologies, Thermo Fisher Scientific). Host and chlamydial DNA were stained with 1 μg/ml 4’, 6-diamidino-2’-phenylindole dihydrochloride (DAPI); Molecular Probes, Thermo Fisher Scientific).…”

Section: Methodsmentioning

confidence: 99%

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Neisseria gonorrhoeae Limits Chlamydia trachomatis Inclusion Development and Infectivity in a Novel In Vitro Co-Infection Model

Onorini

Borel

Schoborg

et al. 2022

Front. Cell. Infect. Microbiol.

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Chlamydia trachomatis (Ct) and Neisseria gonorrhoeae (Ng) are the most common bacterial sexually transmitted infections (STIs) worldwide. The primary site of infection for both bacteria is the epithelium of the endocervix in women and the urethra in men; both can also infect the rectum, pharynx and conjunctiva. Ct/Ng co-infections are more common than expected by chance, suggesting Ct/Ng interactions increase susceptibility and/or transmissibility. To date, studies have largely focused on each pathogen individually and models exploring co-infection are limited. We aimed to determine if Ng co-infection influences chlamydial infection and development and we hypothesized that Ng-infected cells are more susceptible to chlamydial infection than uninfected cells. To address this hypothesis, we established an in vitro model of Ct/Ng co-infection in cultured human cervical epithelial cells. Our data show that Ng co-infection elicits an anti-chlamydial effect by reducing chlamydial infection, inclusion size, and subsequent infectivity. Notably, the anti-chlamydial effect is dependent on Ng viability but not extracellular nutrient depletion or pH modulation. Though this finding is not consistent with our hypothesis, it provides evidence that interaction of these bacteria in vitro influences chlamydial infection and development. This Ct/Ng co-infection model, established in an epithelial cell line, will facilitate further exploration into the pathogenic interplay between Ct and Ng.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Neisseria gonorrhoeae Limits Chlamydia trachomatis Inclusion Development and Infectivity in a Novel In Vitro Co-Infection Model

Onorini

Borel

Schoborg

et al. 2022

Front. Cell. Infect. Microbiol.

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“…For the higher concentration of each pharmaceutic (Mara 20 µM, Cel 1 µM and Az 20 µM), glutaraldehyde fixation and transmission electron microscopy preparation were performed to observe ultrastructural changes of chlamydial inclusions [30, 31]. TEM analyses were performed from two coverslips of one experiment.…”

Section: Methodsmentioning

confidence: 99%

Maraviroc, celastrol and azelastine alter Chlamydia trachomatis development in HeLa cells

Kuratli

Leonard

Nufer

et al. 2020

Journal of Medical Microbiology

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Introduction . Chlamydia trachomatis (Ct) is an obligate intracellular bacterium, causing a range of diseases in humans. Interactions between chlamydiae and antibiotics have been extensively studied in the past. Hypothesis/Gap statement: Chlamydial interactions with non-antibiotic drugs have received less attention and warrant further investigations. We hypothesized that selected cytokine inhibitors would alter Ct growth characteristics in HeLa cells. Aim. To investigate potential interactions between selected cytokine inhibitors and Ct development in vitro. Methodology. The CCR5 receptor antagonist maraviroc (Mara; clinically used as HIV treatment), the triterpenoid celastrol (Cel; used in traditional Chinese medicine) and the histamine H1 receptor antagonist azelastine (Az; clinically used to treat allergic rhinitis and conjunctivitis) were used in a genital in vitro model of Ct serovar E infecting human adenocarcinoma cells (HeLa). Results. Initial analyses revealed no cytotoxicity of Mara up to 20 µM, Cel up to 1 µM and Az up to 20 µM. Mara exposure (1, 5, 10 and 20 µM) elicited a reduction of chlamydial inclusion numbers, while 10 µM reduced chlamydial infectivity. Cel 1 µM, as well as 10 and 20 µM Az, reduced chlamydial inclusion size, number and infectivity. Morphological immunofluorescence and ultrastructural analysis indicated that exposure to 20 µM Az disrupted chlamydial inclusion structure. Immunofluorescence evaluation of Cel-incubated inclusions showed reduced inclusion sizes whilst Mara incubation had no effect on inclusion morphology. Recovery assays demonstrated incomplete recovery of chlamydial infectivity and formation of structures resembling typical chlamydial inclusions upon Az removal. Conclusion. These observations indicate that distinct mechanisms might be involved in potential interactions of the drugs evaluated herein and highlight the need for continued investigation of the interaction of commonly used drugs with Chlamydia and its host.

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“…Confirmation of chlamydial infection usually requires obtaining an appropriate clinical sample from the animal, followed by direct detection of the bacterium using an applicable laboratory diagnostic test: direct impression smears and cytological staining, agent isolation on cell cultures, immunofluorescence tests, enzyme immunoassays, or tests based on nucleic acids amplification [8,2].…”

Section: Introductionmentioning

confidence: 99%

Retrospective analysis of the spread of cat chlamydia and features of its treatment with a drug based on azithromycin

et al. 2020

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The Chlamydiales order bacteria are eukaryotic cells obligate intracellular parasites. The major purpose of the research was to study the prevalence, clinical progression and treatment of Chlamydia in cats with an azithromycin-based drug. A total of 3,388 samples for chlamydia were studied in 2019, of which 243 proved to be positive. Chlamydia was diagnosed using the polymerase chain reaction (PCR) technique. The Azithronit drug was administered 0.5 ml per animal, 1 time per day, for 7 days. Hematological parameters were determined using the IDEXX Laser Cyte hematological analyzer, while the biochemical parameters were determined using the IDEXX Catalist analyzer (USA). In Moscow, chlamydial infection plays a significant role in the cats inflammatory eye diseases etiology. The overall infection rate of the Moscow cat population totalled 7.2%. In young animals, the incidence was higher (28.3%) than in adults (19.5%). In terms of gender, the incidence of chlamydia in female cats (56%) was higher than in males (44%). The main forms of chlamydia manifestations were those of purulent and serous conjunctivitis - 69.8% and 20.5%, respectively, keratitis - 8.7%. The treatment with azithromycin-based drugs requires only a single daily administration for 7 days to ensure the clinical signs elimination.

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The influence of centrifugation and incubation temperatures on various veterinary and human chlamydial species

Cited by 14 publications

References 36 publications

Neisseria gonorrhoeae Limits Chlamydia trachomatis Inclusion Development and Infectivity in a Novel In Vitro Co-Infection Model

Neisseria gonorrhoeae Limits Chlamydia trachomatis Inclusion Development and Infectivity in a Novel In Vitro Co-Infection Model

Maraviroc, celastrol and azelastine alter Chlamydia trachomatis development in HeLa cells

Retrospective analysis of the spread of cat chlamydia and features of its treatment with a drug based on azithromycin

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