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Primary culture of proximal tubular cells (PTC) from normal mouse kidney as an in vitro model to study mechanisms of development of tubulointerstitial nephritis

Research Project

Project/Area Number 05670944
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field Kidney internal medicine
Research InstitutionCHIBA UNIVERSITY

Principal Investigator

UEDA Shiro  CHIBA UNIVERSITY,SCHOOL OF MEDICINE,LECTURER, 医学部, 講師 (50201348)

Co-Investigator(Kenkyū-buntansha) HORI Junro  CHIBA UNIVERSITY,SCHOOL OF MEDICIN,FELLOW, 医学部・附属病院, 医員
OGAWA Makoto  CHIBA UNIVERSITY,CHIBA UNIVERSITY HOSPITAL,ASSISTANT, 医学部・附属病院, 助手 (50241956)
Project Period (FY) 1993 – 1994
Project Status Completed (Fiscal Year 1994)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1994: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1993: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsTubular basement membrane (TBM) anrigen / Cell culture / Proximal tubular cells (PTC) / Tubulointerstitial nephritis / Intercellular adhesion molecule-1 (ICAM-1) / 近位尿細管上皮細胞 / ドーム形成 / ICAM-1
Research Abstract

Cultured murine PTC clearly expressed ICAM-1 under the influence of TBM-primed T lymphocytes. These lymphocytes also demonstrated a high cytotoxic activity against PTC.Purified PTC from BALB/c mice was primary-cultured. PTC was prepared by a modified method of Bogaard et al (Biochem Pharmacol 39 : 13351345,1990). Briefly, kidney was perfused with buffer containing 0.08% (w/v) collagenase. Then, the cortical tissue was filtered through a nylon-gauze. Viable PTC was separated from other materials by isopycnic centrifugation on a discontinuous Nycodenz gradient. The confluent monolayr of PTC showed typical epithelial morphology with cobblestone-like appearanc. Typical dome formation was observed in PTC cultures. These cells also expressed gamma-GTP activity. TBM-primed nylon-column non-adherent lymphocytes were prepared from BALB/c mice immunized with mouse TBM Ag in adjuvant.
Cocultured PTC with TBM-primed lymphocytes induced ICAM-1 expression in PTC.The lymphocytes also had a strong cytotoxic activity without complement. Depletion of T cells from these lymphocytes removed the cytotoxic activity. Neither liver antigen-primed nor DNP-primed lymphocytes had any cytotoxicity against PTC.
This simple syngeneic model may allow further molecularbiological examination of tubulointerstitial nephritis.

Report

(3 results)
  • 1994 Annual Research Report   Final Research Report Summary
  • 1993 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] 堀潤朗他8名: "培養尿細管上皮細胞を用いての間質性腎炎発症機序の解析-尿細管基底膜(TBM)抗原感作リンパ球にBICAM-1の誘導" 日本腎臓学会試. 37(3月25日発行予定). (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 上田志朗: "Annual Review腎臓" 中外医学社, 5 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shiro Ueda: "Clinical Nephrotoxins" Decker,New York, 15 (1995)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Hori J,Ueda S,Anzai N,Itoh K,Ogawa M,Ohto M,Wakashin Y,Wakashin M: "Primary culture of proximal tubular cells(PTC)from normal mouse kidney as an in vitro model to study mechanisms of development of tubulointerstitial nephritis-Induction of ICAM-1 in PTC by antigen-primed lymphocytes." Jap J Nephrol. 37 (in press). (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shiro Ueda: Nephrotoxicity of D-penicillamine, gold salts, and alloprinol.in "Clinical Nephrotoxins" , edt by Bennett WN,De Broe ME,Porter GA,and Varpooten GA.Decker, New York, (in press), (1995)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] Shiro Ueda: Gold nephropathy, in "Annual Review Jinzo 1994" , edt by Kosikawa S,Nagasawa T,Koiso K,Itoh T.CHUGAI IGAKU Co.Tokyo, 102-106 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1994 Final Research Report Summary
  • [Publications] 堀潤朗(他7名): "培養尿細管上皮細胞を用いての間質性腎炎の発症機序の解析 -尿細管基底膜(TBM)抗原感作リンパ球によるICAM-1の誘導-" 日本腎臓学会誌. 37(発行予定). (1995)

    • Related Report
      1994 Annual Research Report
  • [Publications] 上田志郎: "Annual Review腎臓" 中外医学社, 5 (1994)

    • Related Report
      1994 Annual Research Report
  • [Publications] Shiro Ueda: "Clinical Nephrotoxins" Decker,New York, 15 (1995)

    • Related Report
      1994 Annual Research Report

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Published: 1993-04-01   Modified: 2016-04-21  

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