Dataset Information

Proteomic analysis of bovine mammary epithelial cells after in vitro incubation with S. agalactiae: potential biomarkers

ABSTRACT: Streptococcus agalactiae is one of the causative agents of subclinical mastitis, a common disease of dairy cows that causes great economic losses in the industry worldwide. It is thought that pathology is mainly due to infammatory damage of bovine mammary epithelial cells (bMECs); however, the mechanism by which S. agalactiae damages the bMECs is not clear. The aim of this study was to evaluate the infammatory efects of S. agalactiae on bMECs and the resulting changes in protein profles. The bMECs were incubated with S. agalactiae for diferent times and assayed for cell viability by MTT assay, apoptosis by annexin V and propidium iodide dual staining, and morphological and ultrastructural changes by scanning and transmission electron microscopy. Quantitative real-time PCR was used to determine the efect of S. agalactiae on expression of mRNA of infammatory factors in bMECs and protein levels were quantitated by liquid chromatography/mass spectrometry. Exposure to S. agalactiae signifcantly decreased the cell viability and triggered apoptosis, as well as up-regulating TNF-α, IL-1β and IL-6 mRNA, and inhibiting IL-8 expression. S. agalactiae also induced morphological and ultrastructural changes. Furthermore, we identifed 325 up-regulated and 704 down-regulated proteins in the treated vs control group. All signifcant diferentially expressed proteins (DSEPs) were classifed into three major areas by function: biological processes, cellular components and molecular functions. These diferentially expressed proteins included enzymes and proteins associated with various metabolic processes and cellular immunity. Pathway enrichment analysis showed that eight down-regulated signaling pathways were signifcantly enriched. Exposure to even subclinical levels of S. agalactiae can lead to infammation and bMEC damage. Our data suggest some possible molecular mechanisms for the harmful efects of subclinical mastitis in dairy cows.

INSTRUMENT(S): timsTOF Pro

ORGANISM(S): Bos Taurus (bovine)

TISSUE(S): Cell Culture, Mammary Gland Epithelial Cell

DISEASE(S): Breast Cancer

SUBMITTER: Xin Li

LAB HEAD: Jinjin Tong

PROVIDER: PXD028585 | Pride | 2022-10-13

REPOSITORIES: Pride

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Publications

The Analysis of the Ubiquitylomic Responses to <i>Streptococcus agalactiae</i> Infection in Bovine Mammary Gland Epithelial Cells.

Tong Jinjin J Ji Xintong X Zhang Hua H Xiong Benhai B Cui Defeng D Jiang Linshu L

Journal of inflammation research 20220727

<h4>Purpose</h4><i>Streptococcus agalactiae</i> is one of the primary pathogens responsible for subclinical mastitis, a significant economic burden for dairy farms. An essential component of the immune response to infection is ubiquitination, which plays important roles in the complex interactions between the pathogen and host.<h4>Materials and methods</h4>In the present study, quantitative ubiquitylomics was performed to profile changes in the global ubiquitinome of bovine mammary gland epithel ...[more]

PMID: 35923910

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Project description:Bovine mastitis, the infection of the mammary gland which leads to great health and economic challenges for dairy farmers is accompanied by dramatic changes in the milk proteome. In this study of naturally occurring mastitis not only have the changes in the milk proteome been quantified in subclinical and clinical mastitis but simultaneous changes in the serum proteome have also been characterised and quantified. Milk and serum samples from healthy dairy cows (n=12) were compared to those of cows with subclinical (n=10) and clinical mastitis (n=112) using TMT label-based proteomic approach. The study included the milk and serum samples taken from thirty-two dairy cows ( kept on private farms located in Croatia. All cows were checked by physical examination. Somatic cells count (SCC) and mastitis test in milk samples were performed. According to the results, cows were assigned into three groups: Group I (control, n=10) consisted of healthy cows with SCC below 400,000 cells/ml on the monthly check-up and a negative mastitis test and without any clinical sign of mastitis. Group II (subclinical mastitis, n=12) comprised cows without clinical signs of mastitis but with SCC above 400,000 cells/ml on the monthly basis and a positive mastitis test at the time of sampling. Group III (clinical mastitis, n=10) consisted of cows with clinical signs of mastitis which include changes in milk appearance (flakes and clots in milk), different stages of udder inflammation (hyperemia, edema, pain, udder enlargement and elevated udder temperature) and disturbance of general health (depression, relaxed cold ears, dehydration, elevated body temperature, increased heart and respiratory rate, decreased ruminal contraction and decreased appetite). Blood samples were taken from v. coccygea and centrifuged at 3000 g for 15 min after clotting for two hours at room temperature. Serum samples were stored at -80°C until analysis. Milk samples were taken aseptically before the morning milking. First few streams were discarded. Teat ends were disinfected with cotton swabs soaked with 70% ethanol. Samples were taken into sterile tubes and transported to laboratory on ice within a few hours.

Dataset Information

Proteomic analysis of bovine mammary epithelial cells after in vitro incubation with S. agalactiae: potential biomarkers

Publications

The Analysis of the Ubiquitylomic Responses to <i>Streptococcus agalactiae</i> Infection in Bovine Mammary Gland Epithelial Cells.

Similar Datasets

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