Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in Goats
Mastitis results in great economic loss to the dairy goat industry. Many approaches have attempted to decrease the morbidity associated with this disease, and among these, transgenic strategy have been recognized as a potential approach. A previous mammalian study reports that interferon-gamma (IFN-...
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| Format: | Article |
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Frontiers Media S.A.
2022-06-01
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| Series: | Frontiers in Veterinary Science |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fvets.2022.898635/full |
| _version_ | 1811236020372373504 |
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| author | Ying Liu Hongyan Zhang Shasha Dong Boyu Li Weiming Ma Lijiang Ge Zhiyong Hu Feng Su |
| author_facet | Ying Liu Hongyan Zhang Shasha Dong Boyu Li Weiming Ma Lijiang Ge Zhiyong Hu Feng Su |
| author_sort | Ying Liu |
| collection | DOAJ |
| description | Mastitis results in great economic loss to the dairy goat industry. Many approaches have attempted to decrease the morbidity associated with this disease, and among these, transgenic strategy have been recognized as a potential approach. A previous mammalian study reports that interferon-gamma (IFN-γ) has potential anti-bacterial bioactivity against infection in vitro; however, its capacity in vivo is ambiguous. In this study, we initially constructed targeting and homologous recombination vectors (containing the IFN-γ gene) and then transferred the vectors into goat mammary gland epithelial cells (GMECs). Enzyme digestion and sequencing analysis indicated that the vectors used in this study were built correctly. Subsequently, monoclonal cells were selected using puromycin and the polymerase chain reaction (PCR) test indicated that IFN-γ was correctly inserted downstream of the casein promoter. Monoclonal cells were then assessed for reducible expression, and reverse transcriptase-PCR (RT-PCR) and Western blot tests confirmed that monoclonal cells could express IFN-γ. Finally, anti-bacterial capacity was evaluated using bacterial counts and flow cytometry analysis. Decreased bacterial counts and cell apoptosis rates in transgenic GMECs demonstrated that the secretion of IFN-γ could inhibit bacterial proliferation. Therefore, IFN-γ gene transfection in goat mammary epithelial cells could inhibit bacterial proliferation and reduce the risk of mammary gland infection in goats. |
| first_indexed | 2024-04-12T12:02:19Z |
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| id | doaj.art-31c73221d4a64601bfd2e49859f38e91 |
| institution | Directory Open Access Journal |
| issn | 2297-1769 |
| language | English |
| last_indexed | 2024-04-12T12:02:19Z |
| publishDate | 2022-06-01 |
| publisher | Frontiers Media S.A. |
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| spelling | doaj.art-31c73221d4a64601bfd2e49859f38e912022-12-22T03:33:48ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692022-06-01910.3389/fvets.2022.898635898635Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in GoatsYing Liu0Hongyan Zhang1Shasha Dong2Boyu Li3Weiming Ma4Lijiang Ge5Zhiyong Hu6Feng Su7Shandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science, Shandong Agricultural University, Taian, ChinaShandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science, Shandong Agricultural University, Taian, ChinaDepartment of Cardiology, The Second Affiliated Hospital of Shandong First Medical University, Taian, ChinaShandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science, Shandong Agricultural University, Taian, ChinaShandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science, Shandong Agricultural University, Taian, ChinaShandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science, Shandong Agricultural University, Taian, ChinaShandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science, Shandong Agricultural University, Taian, ChinaShandong Provincial Key Laboratory of Animal Biotechnology and Disease Control and Prevention, College of Animal Science, Shandong Agricultural University, Taian, ChinaMastitis results in great economic loss to the dairy goat industry. Many approaches have attempted to decrease the morbidity associated with this disease, and among these, transgenic strategy have been recognized as a potential approach. A previous mammalian study reports that interferon-gamma (IFN-γ) has potential anti-bacterial bioactivity against infection in vitro; however, its capacity in vivo is ambiguous. In this study, we initially constructed targeting and homologous recombination vectors (containing the IFN-γ gene) and then transferred the vectors into goat mammary gland epithelial cells (GMECs). Enzyme digestion and sequencing analysis indicated that the vectors used in this study were built correctly. Subsequently, monoclonal cells were selected using puromycin and the polymerase chain reaction (PCR) test indicated that IFN-γ was correctly inserted downstream of the casein promoter. Monoclonal cells were then assessed for reducible expression, and reverse transcriptase-PCR (RT-PCR) and Western blot tests confirmed that monoclonal cells could express IFN-γ. Finally, anti-bacterial capacity was evaluated using bacterial counts and flow cytometry analysis. Decreased bacterial counts and cell apoptosis rates in transgenic GMECs demonstrated that the secretion of IFN-γ could inhibit bacterial proliferation. Therefore, IFN-γ gene transfection in goat mammary epithelial cells could inhibit bacterial proliferation and reduce the risk of mammary gland infection in goats.https://www.frontiersin.org/articles/10.3389/fvets.2022.898635/fullIFN-γmastitisgene-editinganti-mastitis activityinfection risk |
| spellingShingle | Ying Liu Hongyan Zhang Shasha Dong Boyu Li Weiming Ma Lijiang Ge Zhiyong Hu Feng Su Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in Goats Frontiers in Veterinary Science IFN-γ mastitis gene-editing anti-mastitis activity infection risk |
| title | Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in Goats |
| title_full | Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in Goats |
| title_fullStr | Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in Goats |
| title_full_unstemmed | Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in Goats |
| title_short | Secretion of IFN-γ by Transgenic Mammary Epithelial Cells in vitro Reduced Mastitis Infection Risk in Goats |
| title_sort | secretion of ifn γ by transgenic mammary epithelial cells in vitro reduced mastitis infection risk in goats |
| topic | IFN-γ mastitis gene-editing anti-mastitis activity infection risk |
| url | https://www.frontiersin.org/articles/10.3389/fvets.2022.898635/full |
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