1994 Fiscal Year Final Research Report Summary
Primary culture of proximal tubular cells (PTC) from normal mouse kidney as an in vitro model to study mechanisms of development of tubulointerstitial nephritis
| Project/Area Number |
05670944
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
Kidney internal medicine
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| Research Institution | CHIBA UNIVERSITY |
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Principal Investigator |
UEDA Shiro CHIBA UNIVERSITY,SCHOOL OF MEDICINE,LECTURER, 医学部, 講師 (50201348)
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| 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)
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| Project Period (FY) |
1993 – 1994
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| Keywords | Tubular basement membrane (TBM) anrigen / Cell culture / Proximal tubular cells (PTC) / Tubulointerstitial nephritis / Intercellular adhesion molecule-1 (ICAM-1) |
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| 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.
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