Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells
Abstract Background Innate immune responses induced by in vitro stimulation of primary mammary epithelial cells (MEC) using Gram-negative lipopolysaccharide (LPS) and Gram-positive lipoteichoic acid (LTA) bacterial cell wall components are well- characterized in bovine species. The objective of the...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2017-04-01
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| Series: | Journal of Animal Science and Biotechnology |
| Subjects: | |
| Online Access: | http://link.springer.com/article/10.1186/s40104-017-0162-8 |
| _version_ | 1819119992386355200 |
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| author | Omar Bulgari Xianwen Dong Alfred L. Roca Anna M. Caroli Juan J. Loor |
| author_facet | Omar Bulgari Xianwen Dong Alfred L. Roca Anna M. Caroli Juan J. Loor |
| author_sort | Omar Bulgari |
| collection | DOAJ |
| description | Abstract Background Innate immune responses induced by in vitro stimulation of primary mammary epithelial cells (MEC) using Gram-negative lipopolysaccharide (LPS) and Gram-positive lipoteichoic acid (LTA) bacterial cell wall components are well- characterized in bovine species. The objective of the current study was to characterize the downstream regulation of the inflammatory response induced by Toll-like receptors in primary goat MEC (pgMEC). We performed quantitative real-time RT-PCR (qPCR) to measure mRNA levels of 9 genes involved in transcriptional regulation or antibacterial activity: Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), prostaglandin-endoperoxide synthase 2 (PTGS2), interferon induced protein with tetratricopeptide repeats 3 (IFIT3), interferon regulatory factor 3 (IRF3), myeloid differentiation primary response 88 (MYD88), nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (NFKB1), Toll interacting protein (TOLLIP), and lactoferrin (LTF). Furthermore, we analyzed 7 cytokines involved in Toll-like receptor signaling pathways: C-C motif chemokine ligand 2 (CCL2), C-C motif chemokine ligand 5 (CCL5), C-X-C motif chemokine ligand 6 (CXCL6), interleukin 8 (CXCL8), interleukin 1 beta (IL1B), interleukin 6 (IL6), and tumor necrosis factor alpha (TNF). Results Stimulation of pgMEC with LPS for 3 h led to an increase in expression of CCL2, CXCL6, IL6, CXCL8, PTGS2, IFIT3, MYD88, NFKB1, and TLR4 (P < 0.05). Except for IL6, and PTGS2, the same genes had greater expression than controls at 6 h post-LPS (P < 0.05). Expression of CCL5, PTGS2, IFIT3, NFKB1, TLR4, and TOLLIP was greater than controls after 3 h of incubation with LTA (P < 0.05). Compared to controls, stimulation with LTA for 6 h led to greater expression of PTGS2, IFIT3, NFKB1, and TOLLIP (P < 0.05) whereas the expression of CXCL6, CXCL8, and TLR4 was lower (P < 0.05). At 3 h incubation with both toxins compared to controls a greater expression (P < 0.05) of CCL2, CCL5, CXCL6, CXCL8, IL6, PTGS2, IFIT3, IRF3, MYD88, and NFKB1 was detected. After 6 h of incubation with both toxins, the expression of CCL2, CXCL6, IFIT3, MYD88, NFKB1, and TLR4 was higher than the controls (P < 0.05). Conclusions Data indicate that in the goat MEC, LTA induces a weaker inflammatory response than LPS. This may be related to the observation that gram-positive bacteria cause chronic mastitis more often than gram-negative infections. |
| first_indexed | 2024-12-22T06:13:35Z |
| format | Article |
| id | doaj.art-3878fb1dbd8240d193c9ff74b9c942cc |
| institution | Directory Open Access Journal |
| issn | 2049-1891 |
| language | English |
| last_indexed | 2024-12-22T06:13:35Z |
| publishDate | 2017-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Animal Science and Biotechnology |
| spelling | doaj.art-3878fb1dbd8240d193c9ff74b9c942cc2022-12-21T18:36:09ZengBMCJournal of Animal Science and Biotechnology2049-18912017-04-018111010.1186/s40104-017-0162-8Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cellsOmar Bulgari0Xianwen Dong1Alfred L. Roca2Anna M. Caroli3Juan J. Loor4Department of Animal Sciences and Division of Nutritional Sciences, University of Illinois at Urbana-ChampaignDepartment of Animal Sciences and Division of Nutritional Sciences, University of Illinois at Urbana-ChampaignDepartment of Animal Sciences and Division of Nutritional Sciences, University of Illinois at Urbana-ChampaignDepartment of Molecular and Translational Medicine, University of BresciaDepartment of Animal Sciences and Division of Nutritional Sciences, University of Illinois at Urbana-ChampaignAbstract Background Innate immune responses induced by in vitro stimulation of primary mammary epithelial cells (MEC) using Gram-negative lipopolysaccharide (LPS) and Gram-positive lipoteichoic acid (LTA) bacterial cell wall components are well- characterized in bovine species. The objective of the current study was to characterize the downstream regulation of the inflammatory response induced by Toll-like receptors in primary goat MEC (pgMEC). We performed quantitative real-time RT-PCR (qPCR) to measure mRNA levels of 9 genes involved in transcriptional regulation or antibacterial activity: Toll-like receptor 2 (TLR2), Toll-like receptor 4 (TLR4), prostaglandin-endoperoxide synthase 2 (PTGS2), interferon induced protein with tetratricopeptide repeats 3 (IFIT3), interferon regulatory factor 3 (IRF3), myeloid differentiation primary response 88 (MYD88), nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 (NFKB1), Toll interacting protein (TOLLIP), and lactoferrin (LTF). Furthermore, we analyzed 7 cytokines involved in Toll-like receptor signaling pathways: C-C motif chemokine ligand 2 (CCL2), C-C motif chemokine ligand 5 (CCL5), C-X-C motif chemokine ligand 6 (CXCL6), interleukin 8 (CXCL8), interleukin 1 beta (IL1B), interleukin 6 (IL6), and tumor necrosis factor alpha (TNF). Results Stimulation of pgMEC with LPS for 3 h led to an increase in expression of CCL2, CXCL6, IL6, CXCL8, PTGS2, IFIT3, MYD88, NFKB1, and TLR4 (P < 0.05). Except for IL6, and PTGS2, the same genes had greater expression than controls at 6 h post-LPS (P < 0.05). Expression of CCL5, PTGS2, IFIT3, NFKB1, TLR4, and TOLLIP was greater than controls after 3 h of incubation with LTA (P < 0.05). Compared to controls, stimulation with LTA for 6 h led to greater expression of PTGS2, IFIT3, NFKB1, and TOLLIP (P < 0.05) whereas the expression of CXCL6, CXCL8, and TLR4 was lower (P < 0.05). At 3 h incubation with both toxins compared to controls a greater expression (P < 0.05) of CCL2, CCL5, CXCL6, CXCL8, IL6, PTGS2, IFIT3, IRF3, MYD88, and NFKB1 was detected. After 6 h of incubation with both toxins, the expression of CCL2, CXCL6, IFIT3, MYD88, NFKB1, and TLR4 was higher than the controls (P < 0.05). Conclusions Data indicate that in the goat MEC, LTA induces a weaker inflammatory response than LPS. This may be related to the observation that gram-positive bacteria cause chronic mastitis more often than gram-negative infections.http://link.springer.com/article/10.1186/s40104-017-0162-8Gene expressionInflammationLactationMastitis |
| spellingShingle | Omar Bulgari Xianwen Dong Alfred L. Roca Anna M. Caroli Juan J. Loor Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells Journal of Animal Science and Biotechnology Gene expression Inflammation Lactation Mastitis |
| title | Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells |
| title_full | Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells |
| title_fullStr | Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells |
| title_full_unstemmed | Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells |
| title_short | Innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells |
| title_sort | innate immune responses induced by lipopolysaccharide and lipoteichoic acid in primary goat mammary epithelial cells |
| topic | Gene expression Inflammation Lactation Mastitis |
| url | http://link.springer.com/article/10.1186/s40104-017-0162-8 |
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