Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken Follicles
The aim of this study was to determine the effects of Toll-like receptor (TLR) ligands on the expression of cytokines in chicken follicular theca and to investigate whether nuclear factor-κB (NFκB) was involved in their expression. The follicular theca was collected from the largest follicle of layi...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Japan Poultry Science Association
2018-10-01
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| Series: | The Journal of Poultry Science |
| Subjects: | |
| Online Access: | https://www.jstage.jst.go.jp/article/jpsa/55/4/55_0170217/_html/-char/en |
| _version_ | 1797816983933157376 |
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| author | Ye Kang Takahiro Nii Naoki Isobe Yukinori Yoshimura |
| author_facet | Ye Kang Takahiro Nii Naoki Isobe Yukinori Yoshimura |
| author_sort | Ye Kang |
| collection | DOAJ |
| description | The aim of this study was to determine the effects of Toll-like receptor (TLR) ligands on the expression of cytokines in chicken follicular theca and to investigate whether nuclear factor-κB (NFκB) was involved in their expression. The follicular theca was collected from the largest follicle of laying hens. In experiment 1, the expression of TLRs in the theca interna and externa was confirmed using RT-PCR. The theca tissues were then incubated with or without Pam3CSK4 (TLR2 ligand), poly I:C (TLR3 ligand), LPS (TLR4 ligand), flagellin (TLR5 ligand), R837 (TLR7 ligand), and CpG-ODN (TLR21 ligand) for 3 h, after which cytokine expression (IL-1β, IL-6, TNFSF15, CXCLi2, IFN-α, and IFN-β) was analyzed by real-time PCR. In experiment 2, the theca tissues were incubated in a medium containing Pam3CSK4, poly I:C, LPS, or CpG-ODN with or without BAY 11-7085 (an inhibitor of NFκB) for 3 h. The results of experiment 1 revealed that all TLRs, namely TLR1 (type 1 and 2), TLR2 (type 1 and 2), 3–5, 7, 15, and 21, were expressed in the follicular theca, although the PCR products of TLR1 (type 2) and TLR21 were faint. Moreover, Pam3CSK4 and LPS upregulated the expression of all detected cytokines, except for IFN-α, whose expression was not upregulated by LPS. Poly I:C upregulated the expression of IL-6, CXCLi2, and IFN-β, while CpG-ODN upregulated IL-1β. Flagellin and R837 did not significantly affect cytokine expression. In experiment 2, the expression of IL-1β, IL-6, CXCLi2 and IFN-β in tissues incubated with LPS was downregulated by BAY 11-7085. These results suggest that the innate immune system, including pattern recognition by TLRs and cytokine synthesis, occur in the theca; whereas, functions for recognition of bacterial patterns is more developed than that of viral ones. |
| first_indexed | 2024-03-13T08:45:43Z |
| format | Article |
| id | doaj.art-0a9c757f962242b69fc7c6443d715cc0 |
| institution | Directory Open Access Journal |
| issn | 1346-7395 1349-0486 |
| language | English |
| last_indexed | 2024-03-13T08:45:43Z |
| publishDate | 2018-10-01 |
| publisher | Japan Poultry Science Association |
| record_format | Article |
| series | The Journal of Poultry Science |
| spelling | doaj.art-0a9c757f962242b69fc7c6443d715cc02023-05-30T01:52:42ZengJapan Poultry Science AssociationThe Journal of Poultry Science1346-73951349-04862018-10-0155428830010.2141/jpsa.0170217jpsaEffects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken FolliclesYe Kang0Takahiro Nii1Naoki Isobe2Yukinori Yoshimura3Graduate School of Biosphere Science, Hiroshima UniversityGraduate School of Biosphere Science, Hiroshima UniversityGraduate School of Biosphere Science, Hiroshima UniversityGraduate School of Biosphere Science, Hiroshima UniversityThe aim of this study was to determine the effects of Toll-like receptor (TLR) ligands on the expression of cytokines in chicken follicular theca and to investigate whether nuclear factor-κB (NFκB) was involved in their expression. The follicular theca was collected from the largest follicle of laying hens. In experiment 1, the expression of TLRs in the theca interna and externa was confirmed using RT-PCR. The theca tissues were then incubated with or without Pam3CSK4 (TLR2 ligand), poly I:C (TLR3 ligand), LPS (TLR4 ligand), flagellin (TLR5 ligand), R837 (TLR7 ligand), and CpG-ODN (TLR21 ligand) for 3 h, after which cytokine expression (IL-1β, IL-6, TNFSF15, CXCLi2, IFN-α, and IFN-β) was analyzed by real-time PCR. In experiment 2, the theca tissues were incubated in a medium containing Pam3CSK4, poly I:C, LPS, or CpG-ODN with or without BAY 11-7085 (an inhibitor of NFκB) for 3 h. The results of experiment 1 revealed that all TLRs, namely TLR1 (type 1 and 2), TLR2 (type 1 and 2), 3–5, 7, 15, and 21, were expressed in the follicular theca, although the PCR products of TLR1 (type 2) and TLR21 were faint. Moreover, Pam3CSK4 and LPS upregulated the expression of all detected cytokines, except for IFN-α, whose expression was not upregulated by LPS. Poly I:C upregulated the expression of IL-6, CXCLi2, and IFN-β, while CpG-ODN upregulated IL-1β. Flagellin and R837 did not significantly affect cytokine expression. In experiment 2, the expression of IL-1β, IL-6, CXCLi2 and IFN-β in tissues incubated with LPS was downregulated by BAY 11-7085. These results suggest that the innate immune system, including pattern recognition by TLRs and cytokine synthesis, occur in the theca; whereas, functions for recognition of bacterial patterns is more developed than that of viral ones.https://www.jstage.jst.go.jp/article/jpsa/55/4/55_0170217/_html/-char/enchicken follicular thecacytokinesnfκbtlr ligands |
| spellingShingle | Ye Kang Takahiro Nii Naoki Isobe Yukinori Yoshimura Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken Follicles The Journal of Poultry Science chicken follicular theca cytokines nfκb tlr ligands |
| title | Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken Follicles |
| title_full | Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken Follicles |
| title_fullStr | Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken Follicles |
| title_full_unstemmed | Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken Follicles |
| title_short | Effects of TLR Ligands on the Expression of Cytokines and Possible Role of NFκB in its Process in the Theca of Chicken Follicles |
| title_sort | effects of tlr ligands on the expression of cytokines and possible role of nfκb in its process in the theca of chicken follicles |
| topic | chicken follicular theca cytokines nfκb tlr ligands |
| url | https://www.jstage.jst.go.jp/article/jpsa/55/4/55_0170217/_html/-char/en |
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