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Analysis of Cultured Spiral Ligament Fibrocyles for revealing immune mechanisms of the inner ear

Research Project

Project/Area Number 11671689
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Otorhinolaryngology
Research InstitutionOita Medical University

Principal Investigator

ICHIMIYA Issei  Oita Medical University, Dept. of Immunology and Allergy (Otolaryngology), Associate Professor, 医学部, 助教授 (70223112)

Co-Investigator(Kenkyū-buntansha) YOSHIDA Kazuhide  Oita Medical University, Dept. of Immunology and Allergy (Otolaryngology), Assistant, 医学部, 助手 (10305055)
SUZUKI Masashi  Oita Medical University, Dept. of Immunology and Allergy (Otolaryngology), Professor, 医学部, 教授 (60211314)
MOGI Goro  Oita Medical University, Vice President, 医学部, 副学長 (20035190)
Project Period (FY) 1999 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2001: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2000: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1999: ¥1,000,000 (Direct Cost: ¥1,000,000)
Keywordsadhesion molecules / cell culture / chemokines / fibrocytes / glucocorticoid / immunocytochemistry / spiral ligament / tumor necrosis factor-α / 内耳 / 蝸牛 / ラセン靭帯 / 線維細胞 / 細胞培養 / ギャップ結合 / コネキシン26 / 免疫組織化学 / ラセン靱帯 / 繊維細胞 / サイトカイン / 接着分子 / RT-PCR / グルココルチコイド / ケモカイン / カルデスモン / S-100タンパク / Na-K-ATPase
Research Abstract

1. An immunocytochemical strategy to classify the three types of fibrocytes found in the murine spiral ligament (SL) was established. A combination of staining for caldesmon, S-100 protein, and Na-K-ATPase was useful for this classification.
2. Murine cochlear lateral wall was dissected for secondary culture. Cultures were positive for caldesmon and S-100 protein, and negative for Na-K-ATPase, suggesting that the cells were type I fibrocytes.
3. Cultures were stimulated with IL-1β or TNF-α, and secretion of various mediators was measured. After stimulation, TNF-α, IL-6, MCP-1, KC, MIP-2, slCAM-1, and VEGF levels were elevated. Secretion of these chemokines and other mediators could induce inflammatory cell movement, which would prolong the inflammatory response.
4. Cultures were stimulated with IL-1β or TNF-α, and expression of various adhesion molecules was investigated. Cultures without stimulation did not show staining for VCAM-1, ICAM-1, or MAdCAM-1. Staining was also absent after stimulation with IL-1β; however, VCAM-1 and ICAM-1 staining was observed after cells were stimulated with TNF-α. RT-PCR analysis showed that both VCAM-1 and ICAM-1 mRNAs were present after fibrocytes were stimulated with TNF-α. The findings suggest that activated fibrocytes may cause inflammatory cells to persist in the SL.
5. Immunostaining for glucocorticoid receptors (GRs) was performed, and GRs were found throughout in the murine inner ear. The SL was among the regions where GRs were strongly expressed.
6. Secretion of various mediators was measured after cultures were stimulated with TNF-α and dexamethasone, and the results were compared with those of cells stimulated with TNF-α alone. Dexamethasone suppressed secretion of IL-6, MCP-1, and KC by fibrocytes, suggesting that glucocorticoids have an anti-inflammatory effect on SL fibrocytes.

Report

(5 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • 2000 Annual Research Report
  • 1999 Annual Research Report
  • Research Products

    (47 results)

All Other

All Publications (47 results)

  • [Publications] 吉田和秀 他: "蝸牛ラセン靭帯線維細胞に及ぼす炎症性サイトカインの影響"耳鼻免疫アレルギー. 17. 192-193 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida K 他: "Effect of proinflammatory cytokines on cultured spiral ligament fivrocytes"Hear Res. 137. 155-159 (1999)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Suko T 他: "Classification and culture of spiral ligament fibrocytes from mice"Hear Res. 140. 137-144 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ichimiya I 他: "Significance of spiral ligament fibrocytes with cochlear inflammation"Int J Pediatr Otorhinolaryngol. 56. 45-51 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 一宮一成 他: "実験的長期中耳炎の蝸牛外側壁に及ぼす影響"耳鼻免疫アレルギー. 18. 46-47 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 一宮一成 他: "ラセン靭帯線維細胞におよぼす炎症性サイトカインの影響"Otol Jpn. 10. 155-158 (2000)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida K 他: "Chemokine production from cultured spiral ligament fibrocytes : the effect of corticosteroids"Recent Advances in Otitis Media. 351-354 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ichimiya I 他: "The effect of middle ear effusion on the cochlear lateral wall"Recent Acvances in Otitis Media. 361-364 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 一宮一成 他: "実験的内耳炎モデルの蝸牛におけるmatrix metalloproteinase発現"耳鼻免疫アレルギー. 19. 15-18 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 吉田和秀 他: "内耳ラセン靭帯線維細胞におけるスフェロイド培養の試み"耳鼻免疫アレルギー. 19. 98-99 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 茂木五郎 他: "内耳アレルギー"CLIENT 21(21世紀耳鼻咽喉科臨床の領域). 18. 211-220 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ohya K 他: "Influence of experimental otitis media on the cochlear lateral wall : in vivo and in vitro studies"Recent Advances in Otitis Media with Effusion. 427-428 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shimazaki T 他: "Localization of glucocorticoid receptors in the murine inner ear"Ann Otol Rhinol Laryngol. 111. 1133-1138 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 吉田和秀 他: "グルココルチコイドによるラセン靱帯線維細胞のサイトカイン分泌抑制"耳鼻免疫アレルギー. 20. 86-87 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 一宮一成 他: "内耳炎とラセン靭帯の関わり"耳鼻免疫アレルギー. 20. 132-133 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ichimiya I 他: "Expression of adhesion molecules by cultured spiral ligament fibrocytes stimulated with proinflammatory cytokines"Ann Otol Rhinol Laryngol. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ichimiya I 他: "Aspects of cochlear lateral wall inflammation"Audiological Medicine. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] 一宮一成: "ラセン靱帯の関わる難聴病態"Otol Jpn. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida K, et al.: "Effect of proinflammatory cytokines on cultured spiral ligament fibrocytes"Hear Res 1999. 137. 155-159 (1999)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Suko T, et al.: "Classification and culture of spiral ligament fibrocytes from mice"Hear Res 2000. 140. 137-144 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Ichimiya I, et al.: "Significance of spiral ligament fibrocytes with cochlear inflammation"Intl J Pediatr Otorhinolaryngol. 56. 45-51 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Icimiya I, et al.: "Effect of proinflammatory cytokines on spiral ligament fibrocytes"Ontol Jpn. 10. 155-158 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Yoshida K, et al.: "Chemokine production from cultured spiral ligament fibrocytes: effect of corticosteroids"

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Shimazaki T, Ichimiya I, et al.: "Localization of glucocorticoid receptors in the murine inner ear"Ann Otol Rhinl Laryngol. 111. 1133-1138 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 一宮一成, 鈴木正志, 茂木五郎: "実験的肺炎球菌性中耳炎蝸牛におけるNa-K-Cl共輸送体の分布"日耳鼻. 105. 533 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] 吉田和秀, 一宮一成: "グルココルチコイドによるラセン靭帯線維細胞のサイトカイン分泌抑制"耳鼻免疫アレルギー. 20. 86-87 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 一宮一成, 吉田和秀: "内耳炎とラセン靱帯の関わり"耳鼻免疫アレルギー. 20. 132-133 (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] Ichimiya I, Yoshida K, et al.: "Expression of adhesion molecules by cultured spiral ligament fibrocytes stimulated with proinflammatory cytokines"Ann Otol Rhinol Laryngol. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 一宮一成: "ラセン靭帯の関わる難聴病態"Otol Jpn. (in press). (2003)

    • Related Report
      2002 Annual Research Report
  • [Publications] 吉田和秀, 一宮一成 他: "内耳ラセン靱帯線維細胞におけるスフェロイド培養の試み"耳鼻免疫アレルギー. 19(2). 98-99 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 一宮一成, 嶋崎敏樹 他: "実験的内耳炎モデルの蝸牛におけるmatrix metalloproteinase発現"耳鼻免疫アレルギー. 19(4). 15-18 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 吉田和秀, 一宮一成 他: "グルココルチコイドによるラセン靭帯線維細胞のサイトカイン分泌抑制"日耳鼻. 104(4). 422-422 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 一宮一成, 鈴木正志 他: "走査電子顕微鏡を用いたラセン靱帯の観察"日耳鼻. 104(4). 424-424 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 一宮一成, 鈴木正志 他: "セロイジン包埋側頭骨切片の走査電子顕微鏡による観察"Otol Jpn. 11(4). 287-287 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 茂木五郎, 一宮一成: "内耳アレルギー"CLIENT21(21世紀耳鼻咽喉科臨床の領域). 18. 211-220 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 嶋崎敏樹,一宮一成 他: "マウス内耳におけるグルココルチコイドレセプターの発現"日耳鼻. 103. 103-103 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 一宮一成,吉田和秀 他: "ラセン靭帯線維細胞における細胞接着分子の発現-培養細胞による検討-"日耳鼻. 103. 452-452 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 一宮一成,前田一彦 他: "実験的長期中耳炎の蝸牛外側壁に及ぼす影響"耳鼻免疫アレルギー. 18. 46-47 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 一宮一成,吉田和秀 他: "ラセン靭帯線維細胞におよぼす炎症性サイトカインの影響"Otol Jpn. 10. 155-158 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] 前田恵子,一宮一成 他: "グルココルチコイドによるラセン靭帯線維細胞のサイトカイン分泌抑制"Otol Jpn. 10. 295-295 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Ichimiya I,Yoshida K, et al.: "Significance of spiral ligament fibrocytes with cochlear inflammation"Int J Pediatr Otorhinolaryngol. 56. 45-51 (2000)

    • Related Report
      2000 Annual Research Report
  • [Publications] Yoshida K,Ichimiya I,et al.: "Effect o proinflammatory cytokines on cultured spiral ligament fibrocytes"Hear Res. 137. 155-159 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 一宮一成,須小 毅,他: "マウスラセン靭帯線維細胞の免疫組織学的分類と培養系の確立"日耳鼻. 102. 516-516 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 吉田和秀,一宮一成,他: "蝸牛ラセン靭帯線維細胞に及ぼす炎症サイトカインの影響"耳鼻免疫アレルギー. 17. 192-193 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] 一宮一成,吉田和秀,他: "炎症性サイトカインのラセン靭帯線維細胞に及ぼす影響"Otel Jpn. 9. 502-502 (1999)

    • Related Report
      1999 Annual Research Report
  • [Publications] Ichimiya I,Suzuki M,et al.: "Age-related changes in the murine cochlear lateral wall"Hear Res. 139. 116-122 (2000)

    • Related Report
      1999 Annual Research Report
  • [Publications] Suko T,Ichimiya I,et al.: "Classification and culture of spiral ligament fibrocytes from mice"Hear Res. 140. 137-144 (2000)

    • Related Report
      1999 Annual Research Report

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Published: 1999-04-01   Modified: 2016-04-21  

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