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Expression of the Proteinase Specialized in Bone Resorption, Cathepsin K, in Granulomatous Inflammation

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Abstract

Background

The cysteine proteinase cathepsin K has aroused intense interest as the main effector in the digestion of extracellular matrix during bone resorption by osteoclasts. The enzyme is not a housekeeping lysosomal hydrolase, but is instead expressed with striking specificity in osteoclasts. In this work, we present evidence for the association of cathepsin K with the granulomatous reaction. Granulomas are inflammatory tissue reactions against persistent pathogens or foreign bodies. We came across cathepsin K while working on Echinococcus granulosus, a persistent tissue-dwelling, cyst-forming parasite that elicits a granulomatous response.

Materials and Methods

The walls of hydatid cysts from infected cattle were solubilized. Strong proteolytic activity was detected in the extracts. The proteinase responsible was purified by anion exchange and gel filtration. The purified protein was subjected to N-terminal sequencing, and its identity further confirmed by Western blotting, with a cathepsin K-specific antibody. The same antibody was used to localize the proteinase in paraffin-embedded sections of the parasite and the local host response.

Results

A proteinase was purified to near homogeneity from hydatid cyst extracts. The enzyme was unequivocally identified as host cathepsin K. Both the proenzyme and the mature enzyme forms were found. Cathepsin K was then immunolocalized both to the parasite cyst wall and to the epithelioid and giant multinucleated cells of the host granulomatous response.

Conclusions

In the granulomatous response to the hydatid cyst, cathepsin K is expressed by epithelioid and giant multinucleated cells. We propose that, by analogy with bone resorption, cathepsin K is secreted by the host in an attempt to digest the persistent foreign body. Both processes, bone resorption and granulomatous reactions, therefore tackle persistent extracellular material (the bone matrix or the foreign body), and utilize specialized cells of the monocytic lineage (osteoclasts or epithelioid/giant cells) secreting cathepsin K as an effector.

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Acknowledgments

We are grateful to Dr. A. Punturieri and Dr. S. J. Weiss, of the Division of Hematology and Oncology, Department of Internal Medicine, University of Michigan Medical Center, Ann Arbor, Michigan, U.S.A., for generously providing us with anticathepsin K antibodies and recombinant human cathepsin K.

We thank Jason Pope and Narinder Dass (Department of Pharmacology, University of Oxford, U.K.) for expert help with histochemical stains and with the photography of these, respectively.

We thank Hernán Carol and Silvia González (Cátedra de Inmunología, Facultad de Química, University of Uruguay) for providing us with processed E. granulosus cyst material used in immunohistochemistry.

We thank Miss B. Moffatt for expert technical assistance.

This work was funded by the European Commission’s Directorate General XII/B International Scientific Cooperation programme through a grant (CI1*-CT93-0307) for a joint research effort with the Cátedra de Inmunología, Facultad de Química, University of Uruguay. A.D. was additionally supported by scholarships from the Consejo Nacional de Investigaciones Científicas y Técnicas (CONICYT, Ministry of Education, Uruguay) and the Comisión Sectorial de Investigación Científica (CSIC, University of Uruguay).

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  1. MRC Immunochemistry Unit, Department of Biochemistry, University of Oxford, Oxford, UK

    Alvaro Díaz, Antony C. Willis & Robert B. Sim

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  1. Alvaro Díaz
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Correspondence to Alvaro Díaz.

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Díaz, A., Willis, A.C. & Sim, R.B. Expression of the Proteinase Specialized in Bone Resorption, Cathepsin K, in Granulomatous Inflammation. Mol Med 6, 648–659 (2000). https://doi.org/10.1007/BF03402045

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