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Biologically active ADAMTS13 is expressed in renal tubular epithelial cells

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Abstract

ADAMTS13 mRNA, which encodes the von Willebrand factor-cleaving protease, has been detected in a variety of tissues, including the kidney. The aim of our study was to characterize tubular expression and bioactivity of ADAMTS13. ADAMTS13 mRNA was detected in cultured primary human renal tubular epithelial cells (HRTEC) and in A498 cells, a human renal carcinoma cell line, by real-time PCR. Protein was detected using immunofluorescence and immunoblotting. Immunoblots demonstrated that the protein was secreted. The protease was proteolytically active in both cell lysates and cleaved the FRETS–VWF73 substrate. ADAMTS13 was demonstrated in situ in the renal cortex by immunohistochemistry. Protease was detected in both the proximal and distal renal tubules in normal renal tissue (n = 3) as well as in patients with tubular disorders (n = 3). Immunoblotting revealed that ADAMTS13 was present in the urine of patients with tubulopathy (n = 5) but not in normal urine. ADAMTS13 in urine had a molecular size similar to that in plasma, which indicates that the protease originates in the tubuli because such large proteins do not normally pass the glomerular filter. In conclusion, human renal tubular epithelial cells synthesize biologically active ADAMTS13 which may, after release from tubuli, regulate hemostasis in the local microenvironment.

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Acknowledgments

The authors thank Professor Lars Holmberg (Department of Pediatrics, Lund University, Lund, Sweden) for sharing the SU19 anti-ADAMTS13 polyclonal antibody; Professor Catharina Svanborg and Dr. Majlis Svensson (Department of Microbiology, Immunology, and Glycobiology, Lund University) for the A498 cells; Dr. Andreas Hillarp, Department of Clinical Chemistry, Malmö University Hospital, Malmö, Sweden for assistance in performing the FRETS-VWF73 ADAMTS13 activity assay.

This study was supported by grants from The Swedish Research Council (K2007-64X-14008-07-3), Sven Jerring Foundation, the Crafoord Foundation, The Fund for Renal Research, Crown Princess Lovisa’s Society for Child Care, The Maggie Stephen’s Foundation, The Magnus Bergvall Foundation, The Foundation for Fighting Blood Disease, Konung Gustaf V:s 80-årsfond, and the Fanny Ekdahl Foundation (all to DK). Diana Karpman is the recipient of a clinical-experimental research fellowship from the Royal Swedish Academy of Sciences.

This paper was presented orally at the 3rd International Workshop, Thrombotic Microangiopathies, Jena Germany, October 4–6, 2007. A previous version of the paper appeared in the PhD thesis of Dr. Minola Manea.

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  1. Department of Pediatrics, Clinical Sciences Lund, Lund University, 22185, Lund, Sweden

    Minola Manea, Ramesh Tati, Jessica Karlsson, Zivile D. Békássy & Diana Karpman

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  1. Minola Manea
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  2. Ramesh Tati
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  3. Jessica Karlsson
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  4. Zivile D. Békássy
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  5. Diana Karpman
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Correspondence to Diana Karpman.

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Manea, M., Tati, R., Karlsson, J. et al. Biologically active ADAMTS13 is expressed in renal tubular epithelial cells. Pediatr Nephrol 25, 87–96 (2010). https://doi.org/10.1007/s00467-009-1262-2

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