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Macrophages induce apoptosis in proximal tubule cells

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Abstract.

Macrophages play important roles during renal inflammation. Infiltrating macrophages produce proinflammatory mediators and induce apoptosis in a variety of target cells. Because proximal tubules are frequently damaged in inflammatory processes, we investigated murine macrophages (J774) in the induction of apoptosis in murine PKSV-PR proximal tubule cells. PKSV-PR cells were co-cultured with activated or non-activated macrophages. Apoptosis was assessed by Annexin-V-FITC/propidium iodide staining and analyzed by fluorescence-activated cell sorting. Macrophages were separated from tubule cells with transwell membranes to distinguish soluble factor-mediated from direct cell-to-cell contact-mediated apoptosis. Cell supernatants from activated and non-activated macrophages were analyzed for induction of apoptosis. Activated (but not non-activated) macrophages induced tubule cell apoptosis in co-culture. Soluble factors were mainly responsible for induction of apoptosis; membrane separation and transfer of cell supernatant from activated macrophages showed similar levels of apoptosis induction. Although tumor necrosis factor (TNF)-α and transforming growth factor (TGF)-β1, measured by ELISA, increased significantly in supernatants from activated macrophages, blocking TNF-α and TGF-β did not decrease apoptosis in PKSV-PR cells co-cultured with macrophages. Moreover, exogenous addition of TNF-α, TGF-β, anti-Fas antibody, or TRAIL failed to induce apoptosis in tubule cells. We conclude that inflammatory macrophages mediate proximal tubule cell death, directing apoptosis mainly via release of unidentified soluble factors.

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Acknowledgements.

This research was supported in part by NIDDK grants DK52612 and DK45179 to Dr. Robert L. Chevalier. Dr. Bärbel Lange-Sperandio is supported by a stipend from the German Research Foundation (DFG) La 1257/1–1 and by a Children′s Medical Center (CMC) grant, Charlottesville, University of Virginia.

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Authors and Affiliations

  1. Department of Pediatrics, University of Virginia, Charlottesville, VA 22908, USA

    Bärbel Lange-Sperandio & Robert L. Chevalier

  2. Department of Pediatrics, University of Ulm, 89075 Ulm, Germany

    Simone Fulda

  3. Unité 246, Faculty of Medicine Xavier Bichat , INSERM, BP 416, 75870 Paris, Cedex 18, France

    Alain Vandewalle

  4. Department of Pediatrics, University of Virginia Health Sciences Center, PO Box 800386, Charlottesville, VA 22908, USA

    Robert L. Chevalier

Authors
  1. Bärbel Lange-Sperandio
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  2. Simone Fulda
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  3. Alain Vandewalle
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  4. Robert L. Chevalier
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Correspondence to Robert L. Chevalier.

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Lange-Sperandio, B., Fulda, S., Vandewalle, A. et al. Macrophages induce apoptosis in proximal tubule cells. Pediatr Nephrol 18, 335–341 (2003). https://doi.org/10.1007/s00467-003-1116-2

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  • DOI: https://doi.org/10.1007/s00467-003-1116-2

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