Virus-Mediated Metalloproteinase 1 Induction Revealed by Transcriptome Profiling of Bovine Herpesvirus 4-Infected Bovine Endometrial Stromal Cells.
Viral infections can cause genital tract disorders (including abortion) in cows, and bovine herpesvirus 4 (BoHV-4) is often present in endometritis-affected animals. A major problem with cattle uterine viral infections in general, and BoHV-4 in particular, is our limited understanding of the pathogenic role(s) that these infections play in the endometrium. A similar lack of knowledge holds for the molecular mechanisms utilized, and the host cell pathways affected, by BoHV-4. To begin to fill these gaps, we set up optimized conditions for BoHV-4 infection of a pure population of bovine endometrial stromal cells (BESCs) to be used as source material for RNA sequencing-based transcriptome profiling. Many genes were found to be upregulated (417) or downregulated (181) after BoHV-4 infection. As revealed by enrichment functional analysis on differentially expressed genes, BoHV-4 infection affects various pathways related to cell proliferation and cell surface integrity, at least three of which were centered on upregulation of matrix metalloproteinase 1 (MMP1) and interleukin 8 (IL8). This was confirmed by reverse transcription PCR, real-time PCR, Western-immunoblot analysis, and a luciferase assay with a bovine MMP1-specific promoter reporter construct. Further, it was found that MMP1 transcription was upregulated by the BoHV-4 transactivator IE2/RTA, leading to abnormally high metalloproteinase tissue levels, potentially leading to defective endometrium healing and unresolved inflammation. Based on these findings, we propose a new model for BoHV-4 action centered on IE2-mediated MMP1 upregulation and novel therapeutic interventions based on IFN gamma-mediated MMP1 downregulation.
Department of Medical-Veterinary Science, University of Parma, Parma, Italy.
Giulia
G
Tebaldi
Department of Medical-Veterinary Science, University of Parma, Parma, Italy.
Sarah
S
Jacca
Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Parma, Italy.
Barbara
B
Montanini
National Research Council (CNR), Institute of Agricultural Biology and Biotechnology, Lodi, Italy.
Emanuele
E
Capra
Department of Medical-Veterinary Science, University of Parma, Parma, Italy.
Alfonso
A
Rosamilia
Department of Medical-Veterinary Science, University of Parma, Parma, Italy.
Arianna
A
Sala
National Research Council (CNR), Institute of Agricultural Biology and Biotechnology, Lodi, Italy.
Alessandra
A
Stella
National Research Council (CNR), Institute of Agricultural Biology and Biotechnology, Lodi, Italy.
Bianca
B
Castiglioni
Department of Life Sciences, Biochemistry and Molecular Biology Unit, University of Parma, Parma, Italy.
Simone
S
Ottonello
Department of Medical-Veterinary Science, University of Parma, Parma, Italy gaetano.donofrio@unipr.it.
Gaetano
G
Donofrio
D000818:Animals; D002417:Cattle; D004717:Endometrium / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D005260:Female; D020869:Gene Expression Profiling; D026021:Herpesvirus 4, Bovine; D007249:Inflammation / Q000378:metabolism / Q000473:pathology / Q000821:virology; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000235:genetics / Q000378:metabolism*; D017154:Stromal Cells / Q000378:metabolism* / Q000473:pathology / Q000821:virology; D015854:Up-Regulation
D016209:Interleukin-8; D020781:Matrix Metalloproteinase 1
MMPs; bovine endometrial stromal cells; bovine herpesvirus 4; postpartum metritis; transcriptome profiling