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Responses of bovine endometrial epithelial cells to pathogens

Chanrot, Metasu (2017). Responses of bovine endometrial epithelial cells to pathogens. Diss. (sammanfattning/summary) Uppsala : Sveriges lantbruksuniv., Acta Universitatis agriculturae Sueciae, 1652-6880 ; 2017:16
ISBN 978-91-576-8805-7
eISBN 978-91-576-8806-4
[Doctoral thesis]

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Abstract

In dairy cows, clinical uterine infection (metritis) and subsequent persistent inflammation of the endometrium (endometritis) are major causes of infertility. Escherichia coli (E. coli) is a prevalent bacteria in metritis and endometritis and promotes infection with bovine herpes virus type 4 (BoHV-4) through mechanisms involving lipopolysaccharide endotoxins (LPS). In this thesis, interactions between E. coli LPS, BoHV-4 and the endometrial epithelium were studied using in vitro models following characterisation of tissue samples used for culture. Examination of cell proliferation, survival and apoptosis after challenges with various doses of LPS revealed that cow and tissue characteristics did not influence proliferation of bovine endometrial epithelial cells (bEEC) in response to LPS. However, E. coli LPS stimulated proliferation of bEEC (maximum observed at 8 µg/mL LPS). The strong increase in cell numbers by 72 h was not associated with an increase in apoptosis, but this occurred with higher LPS doses. Analysis of protein pro-files revealed de-regulation of 38 proteins belonging to many pathways, some related to the process of implantation. Morphological studies and ELISA were used to characterise the survival of cells and the cytokine response of bEEC to BoHV-4. In infected samples, the number of living cells started to decrease by Day 4 post-challenge and by Day 7 the number was lower than in controls. This change was associated with viral replication between Day 0 and Day 5, as demonstrated by immunofluorescence, titration and quantitative polymerase chain reaction (qPCR) results and changes in IL-8 and TNF-α profiles. Moreover, the results showing strong pathogenic effects of BoHV-4 on endometrial epithelial cells pave the way for future studies on sexual transmission of BoHV-4 at time of insemination.

The results obtained led to development of reliable models to study interactions between uterine epithelial cells and pathogens, which could be of translational use. In a time- and dose-dependent manner, E. coli LPS increased and BoHV-4 decreased the survival of bovine bEEC in vitro, while LPS induced strong alterations of protein profiles, especially those related to pathways activated at time of implantation. Such de-regulations may be part of the mechanism by which persistent inflammation following infection impairs fertility. This information can be exploited to identify new diagnostic markers of persistence of inflammation in the endometrium.

Authors/Creators:Chanrot, Metasu
Title:Responses of bovine endometrial epithelial cells to pathogens
Series/Journal:Acta Universitatis agriculturae Sueciae (1652-6880)
Year of publishing :2017
Depositing date:2017
Volume:2017:16
Number of Pages:80
Papers/manuscripts:
NumberReferences
IChanrot M*, Guo Y, Dalin AM, Persson E, Båge R, Svensson A, Gustafsson H & Humblot P. (2017). Dose-related effects of LPS on endometrial epithelial cell populations from dioestrous cows. Animal Reproduction Science, 177, pp. 12-24.
IIPiras C, Guo Y, Soggiu A, Chanrot M, Greco V, Urbani A, Charpigny G, Bonizzi L, Roncada P* & Humblot P. (2017). Changes in protein expression profiles in bovine endometrial epithelial cells exposed to E. coli LPS challenge. Molecular BioSystems, 13(2), pp. 392-405.
IIIChanrot M*, Blomqvist G, Guo Y, Ullman K, Juremalm M, Båge R, Valarcher J-F, Donofrio G & Humblot P. Bovine herpesvirus type 4 (BoHV-4) impairs the survival of bovine endometrial epithelial cells (bEEC). (submitted)
Place of Publication:Uppsala
Publisher:Department of Clinical Sciences, Swedish University of Agricultural Sciences
ISBN for printed version:978-91-576-8805-7
ISBN for electronic version:978-91-576-8806-4
ISSN:1652-6880
Language:English
Publication Type:Doctoral thesis
Full Text Status:Public
Agris subject categories.:L Animal production > L53 Animal physiology - Reproduction
L Animal production > L73 Animal diseases
Subjects:(A) Swedish standard research categories 2011 > 3 Medical and Health Sciences > 302 Clinical Medicine > Obstetrics, Gynaecology and Reproductive Medicine
(A) Swedish standard research categories 2011 > 3 Medical and Health Sciences > 304 Medical Biotechnology > Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
(A) Swedish standard research categories 2011 > 4 Agricultural Sciences > 403 Veterinary Science > Clinical Science
Agrovoc terms:dairy cows, endometrium, endometritis, epithelium, cell culture, oestrous cycle, inflammation, pathogens, in vitro experimentation, laboratory diagnosis
Keywords:cow, LPS, BoHV-4, endometrium, cell culture, proteomics, cell proliferation, endometritis, oestrous cycle
URN:NBN:urn:nbn:se:slu:epsilon-e-3998
Permanent URL:
http://urn.kb.se/resolve?urn=urn:nbn:se:slu:epsilon-e-3998
ID Code:14124
Faculty:VH - Faculty of Veterinary Medicine and Animal Science
Department:(VH) > Dept. of Clinical Sciences
External funders:Rajamangala University of Technology Srivijaya
Deposited By: Miss Metasu Chanrot
Deposited On:02 Mar 2017 12:27
Metadata Last Modified:06 Mar 2017 08:57

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