半定量的RT-PCR法を用いたマウス網膜における抗原提示機能の可能性と解析

原, 宏二, 亀谷, 修平, 山木, 邦比古

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The use of semi-quantitative Reverse Transcription-Polymerase Chain Reaction (RT-PCR) to analyze antigen-presenting functions in mouse retina
半定量的RT-PCR法を用いたマウス網膜における抗原提示機能の可能性の解析
ハンテイリョウテキ RT PCRホウオモチイタマウスモウマクニオケルコウゲンテイジキノウノカノウセイノカイセキ

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To identify the possibility of antigen presentation in the retina, we studied the production of various cytokines and the expression of cell surface molecules in the retina by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Lipopolysaccharide (LPS) or inactivated Bordetella pertussis (BP) was injected into the vitreous cavity of C57BL/6J mice. The retinas were collected 3,6 and 24 hours after the injection. Expression of mRNA for MHC class II, CDllc, CDSO, CDS6, toll-like receptor (TLR) 4, TLR9, IL-12p35, IL-2, IFN-y, IL-4, IL-5, IL-6, IL-IO, IL-lj3, TNF-a and TGF-j3 was analyzed by semi-quantitative RT-PCR. Expression of mRNA for MHC class II, CDllc, CD80, CDS6, TLR4 and TLR9 was found in the untreated retina. The amount of expression of MHC class II, CDllc, CDSO and CD86 increased after injection of either LPS or BP. Expression of TLR4 mRNA increased only following injection with LPS, whereas TLR9 increased following injection with BP or LPS, not clear. The expression of cytokines, IL-12p35, IL-2, IFN-y, IL-4, IL-5, IL-6, IL-IO, IL-lj3, TNF-a and TGF-j3 increased after injection with either LPS or BP. A time course of mRNA expression indicated a peak at 3 hours after injection, which then gradually subsided. These results suggest that proinflammatory cytokines related to antigen presentation were upregulated by stimulation of TLR4 and TLR9.

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秋田医学

秋田医学 31 (2), 141-152, 2004-09-01

秋田医学会

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