Studies to Demonstrate Inhibition of Functional Activity of Neutrophil Lysosomal Enzymes with ANCA

  • Linus Chang
  • Judy Savige
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 336)

Summary

Many autoantibodies that have enzymes as their targets bind to the catalytic sites and inhibit enzymatic activity (1). We have established functional assays for the targets of anti-neutrophil cytoplasm antibodies (ANCA) in order to determine whether these antibodies are directed against the corresponding catalytic sites. Anti-proteinase 3 activity was measured by the cleavage of alpha-naphthyl acetate; myeloperoxidase activity by peroxidation of monochlorodimedon or guaiacol using hydrogen peroxide; and elastase activity by a fluorimetric assay of the hydrolytic product of N-succ (ala)3 amido-methyl coumarin. The addition of immunoglobulin from patients with ANCA to these assays did not result in inhibition of functional activity of the corresponding enzyme when compared with normal immunoglobulin. Furthermore the removal of specific immunoglobulin by solid phase adsorption to the corresponding antigen did not result in an increase in enzymatic activity compared with the starting material. However preliminary results suggest that anti-neutrophil proteinase 3 binding may be inhibited in a solid-phase ELISA by preincubation with a peptide corresponding to the catalytic site.

Keywords

Catalytic Site Functional Assay Myeloperoxidase Activity Elastase Activity Fluorimetric Assay 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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References

  1. 1.
    Tan, E.M., 1989, Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology, Adv Immunol. 44: 93–151.PubMedCrossRefGoogle Scholar
  2. 2.
    Kao, R.C., Wehmer,.G., Skubitz, K.M., Gray, B.H. and Hoidal, J.R., 1988, Proteinase 3. A distinct human polymorphonuclear leukocyte proteinase that produces emphysema in hamsters, J Clin Invest. 82: 1963–1973.Google Scholar
  3. 3.
    Lüdeman, J., Csernok, E., Ulmer, M., Lemke, H., Utecht, B., Rautman, A., and Gross, W.1., 1991, Anti-neutrophil cytoplasm antibodies in Wegener’s granulomatosis. immunodiagnostic value, monoclonal antibodies and characterisation of the target antigen, Netherlands J Med. 36: 157162.Google Scholar
  4. 4.
    Lesavre, P., 1991, Anti-neutrophil cytoplasmic autoantibodies antigen specificity, Am J Kid Diseases, 18: 159–163.Google Scholar
  5. 5.
    Jenne, D.E., Tscopp, J., Lüdeman, J., Utecht, B., and Gross, W.L., 1990, Wegener’s autoantigen decoded, Nature, 346: 520.PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1993

Authors and Affiliations

  • Linus Chang
    • 1
  • Judy Savige
    • 2
  1. 1.Department of HaematologyHeidelberg Repatriation HospitalVictoriaAustralia
  2. 2.Department of MedicineAustin Hospital, HeidelbergVictoriaAustralia

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