Dynamic comparison of Histophilus somni Outer Membrane Vesicles (OMVs)

Date

2023-10-13

Author

Aktaş, Caner
Okay, Sezer
Özcan, Servet
Ayaz-Güner, Şerife
Özcengiz, Gülay

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Aim: Histophilus somni is a clinicaly important gram-negative pathogen that causes thromboembolic meningoencephalitis, pneumonia, polyarthritis and acute myocarditis in cattle, as well as infections of the urogenital tract and abortions and fetal septicemia in pregnant animals. Outer membrane vesicles (OMVs) are non-replicating and highly immunogenic spherical nanoparticles derived by Gram-negative bacteria. These particles effectively transmit virulence factors to the host, increasing pathogenesis and activating antigen-presenting host cells. In this project, the immunogenic and protective potentials of OMVs is investigated with the proteomic and RNA sequenscing analyzes. Methods: H. somni standart strain ATCC 43625 have been cultured in 5% sheep blood agar for 24 hours twice and transferred to Brain heart Infusion Broth supplemented with 0.1% tris-base, and 0.001% thiamine hydrochloride. Liquid culture samples are collected at the different stages of logaritmic growth phase and cells were pelleted. Acquired supernatant is subjected to series of ultracentrifugation to isolate OMVs. Concentration and size of OMVs analyzed with Nanoparticle Tracking analyses(NTA) and sample concentrations are equalized. Proteins are purified from whole OMV samples and identified by Liquid Chromatography-Tandem Mass Spectrometry. Similarly RNAs are isolated from whole OMVs with affinity column based purification method and single-read sequencing is conducted to acquire small RNA libraries.Results: It is found that OMVs are packed with many of the known surface antigens as well as structural proteins and porins with varying expression levels. similarly the unique small RNA content that are found in OMVs differs in terms of relative expression through different stages of logaritmic growth.Conclusion: With this dynamic comparison study it is concluded that the OMV contents show high similarity to the antigenic surface of the pathogen. However content of OMVs changes during the logaritmic growth of the bacteria in terms of relative abundance. Therefore it is important to consider the sample collection time.

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https://proteomikdernegi.org/tr/26/352/0/davet.html#davet
https://hdl.handle.net/11511/112457

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Department of Biology, Conference / Seminar