Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis
AIM: To investigate the expression of macrophage migration inhibitory factor (MIF) and detect its role in the innate immune response of fungal keratitis (FK). METHODS: We collected the paraffin-embedded cornea tissues from 10 FK and 6 ocular trauma patients to explore the MIF expression by immunohi...
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| Format: | Article |
| Language: | English |
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Press of International Journal of Ophthalmology (IJO PRESS)
2019-05-01
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| Series: | International Journal of Ophthalmology |
| Subjects: | |
| Online Access: | http://www.ijo.cn/en_publish/2019/5/20190503.pdf |
| _version_ | 1818321310495801344 |
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| author | Qiang Xu Li-Ting Hu Qian Wang Jing Lin Nan Jiang Cui Li Gui-Qiu Zhao |
| author_facet | Qiang Xu Li-Ting Hu Qian Wang Jing Lin Nan Jiang Cui Li Gui-Qiu Zhao |
| author_sort | Qiang Xu |
| collection | DOAJ |
| description | AIM: To investigate the expression of macrophage migration inhibitory factor (MIF) and detect its role in the innate immune response of fungal keratitis (FK).
METHODS: We collected the paraffin-embedded cornea tissues from 10 FK and 6 ocular trauma patients to explore the MIF expression by immunohistochemistry. Then we cultured telomease-immortalized human corneal epithelial cells (THCEs), stimulated by the hyphae suspension of Aspergillus fumigatus (A. fumigatus) to detect the change of MIF with or without the pretreatment of MIF inhibitor [4-Iodo-6-phenylpyrimidine (4-IPP)] by real-time polymerase chain reaction (PCR). The protein level of MIF was also tested by immunohistochemistry, and the level of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were compared between normal, hyphae stimulated and 4-IPP pretreated groups by real-time PCR to study the influence of MIF on the expression of TNF-α and IL-6. Corneal severity of rats’ FK models was documented by clinical scores, and real-time PCR. Western blot and immunohistochemistry were used to test the expression of MIF, TNF-α and IL-6 in rats’ corneas.
RESULTS: In the corneas of FK patients, there was much stronger expression of MIF than that in the normal group showed by immunohistochemistry. In cultured THCEs stimulated by A. fumigatus, the expression of MIF became stronger in both immunohistochemistry and PCR at 16, 24, 32 and 48h post infection (p.i.; P<0.01, P<0.01, P<0.01, P<0.05). After pretreated with 4-IPP, the expression of MIF reduced at 4, 8, 16h p.i. (P<0.05, P<0.05, P<0.05) and the downstream TNF-α and IL-6 decreased obviously (P<0.05, P<0.01). In rats with A. fumigatus keratitis, the relative mRNA and protein level of MIF increased than those in the normal group by PCR (at 1d: P<0.01, 3d: P<0.01, 5d: P<0.01), Western blot and immunohistochemistry. After blocked MIF with 4-IPP, the clinical outcomes of rat keratitis showed markedly reduced inflammatory response (P<0.01), with TNF-α and IL-6 decreased in accordance with those in THCEs by PCR (P<0.05, P<0.01).
CONCLUSION: The expression of MIF increased significantly in FK patients, THCEs and rats stimulated by A. fumigatus. After blocked with 4-IPP, the expression of MIF reduced, and so did its downstream cytokines: TNF-α and IL-6. The inflammation reaction of the rats’ corneas lightened after pretreated with 4-IPP. MIF may play a role in the innate immune response of the corneal resistance against A. fumigatus. |
| first_indexed | 2024-12-13T10:38:52Z |
| format | Article |
| id | doaj.art-307480808dd1440787fc69a7ee73e78c |
| institution | Directory Open Access Journal |
| issn | 2222-3959 2227-4898 |
| language | English |
| last_indexed | 2024-12-13T10:38:52Z |
| publishDate | 2019-05-01 |
| publisher | Press of International Journal of Ophthalmology (IJO PRESS) |
| record_format | Article |
| series | International Journal of Ophthalmology |
| spelling | doaj.art-307480808dd1440787fc69a7ee73e78c2022-12-21T23:50:37ZengPress of International Journal of Ophthalmology (IJO PRESS)International Journal of Ophthalmology2222-39592227-48982019-05-0112571171610.18240/ijo.2019.05.03Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitisQiang Xu0Li-Ting Hu1Qian Wang2Jing Lin3Nan Jiang4Cui Li5Gui-Qiu Zhao6Department of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, ChinaDepartment of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, ChinaDepartment of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, ChinaDepartment of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, ChinaDepartment of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, ChinaDepartment of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, ChinaDepartment of Ophthalmology, the Affiliated Hospital of Qingdao University, Qingdao 266000, Shandong Province, ChinaAIM: To investigate the expression of macrophage migration inhibitory factor (MIF) and detect its role in the innate immune response of fungal keratitis (FK). METHODS: We collected the paraffin-embedded cornea tissues from 10 FK and 6 ocular trauma patients to explore the MIF expression by immunohistochemistry. Then we cultured telomease-immortalized human corneal epithelial cells (THCEs), stimulated by the hyphae suspension of Aspergillus fumigatus (A. fumigatus) to detect the change of MIF with or without the pretreatment of MIF inhibitor [4-Iodo-6-phenylpyrimidine (4-IPP)] by real-time polymerase chain reaction (PCR). The protein level of MIF was also tested by immunohistochemistry, and the level of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) mRNA were compared between normal, hyphae stimulated and 4-IPP pretreated groups by real-time PCR to study the influence of MIF on the expression of TNF-α and IL-6. Corneal severity of rats’ FK models was documented by clinical scores, and real-time PCR. Western blot and immunohistochemistry were used to test the expression of MIF, TNF-α and IL-6 in rats’ corneas. RESULTS: In the corneas of FK patients, there was much stronger expression of MIF than that in the normal group showed by immunohistochemistry. In cultured THCEs stimulated by A. fumigatus, the expression of MIF became stronger in both immunohistochemistry and PCR at 16, 24, 32 and 48h post infection (p.i.; P<0.01, P<0.01, P<0.01, P<0.05). After pretreated with 4-IPP, the expression of MIF reduced at 4, 8, 16h p.i. (P<0.05, P<0.05, P<0.05) and the downstream TNF-α and IL-6 decreased obviously (P<0.05, P<0.01). In rats with A. fumigatus keratitis, the relative mRNA and protein level of MIF increased than those in the normal group by PCR (at 1d: P<0.01, 3d: P<0.01, 5d: P<0.01), Western blot and immunohistochemistry. After blocked MIF with 4-IPP, the clinical outcomes of rat keratitis showed markedly reduced inflammatory response (P<0.01), with TNF-α and IL-6 decreased in accordance with those in THCEs by PCR (P<0.05, P<0.01). CONCLUSION: The expression of MIF increased significantly in FK patients, THCEs and rats stimulated by A. fumigatus. After blocked with 4-IPP, the expression of MIF reduced, and so did its downstream cytokines: TNF-α and IL-6. The inflammation reaction of the rats’ corneas lightened after pretreated with 4-IPP. MIF may play a role in the innate immune response of the corneal resistance against A. fumigatus.http://www.ijo.cn/en_publish/2019/5/20190503.pdfmacrophage migration inhibitory factorfungal keratitisinnate immunea. fumigatuscorneal epithelial cellsrats |
| spellingShingle | Qiang Xu Li-Ting Hu Qian Wang Jing Lin Nan Jiang Cui Li Gui-Qiu Zhao Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis International Journal of Ophthalmology macrophage migration inhibitory factor fungal keratitis innate immune a. fumigatus corneal epithelial cells rats |
| title | Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis |
| title_full | Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis |
| title_fullStr | Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis |
| title_full_unstemmed | Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis |
| title_short | Expression of macrophage migration inhibitory factor in Aspergillus fumigatus keratitis |
| title_sort | expression of macrophage migration inhibitory factor in aspergillus fumigatus keratitis |
| topic | macrophage migration inhibitory factor fungal keratitis innate immune a. fumigatus corneal epithelial cells rats |
| url | http://www.ijo.cn/en_publish/2019/5/20190503.pdf |
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