Pathway-specific complement activity in pigs evaluated with a human functional complement assay

Mol Immunol. 2009 May;46(8-9):1620-5. doi: 10.1016/j.molimm.2009.02.028. Epub 2009 Mar 27.

Abstract

Background: The complement system is an important part of innate immunity. Complement deficiencies or inappropriate activation of complement may cause severe diseases. The complement functional test, Wielisa, assesses all three complement activation pathways in humans. It is important to have assays available to determine the functional complement activity in research animals. Since the pig is a relevant animal in experimental research, the aim of the present study was to evaluate the applicability of this human complement assay in pigs.

Methods: Normal pig serum was serially diluted and assayed in the Wielisa test which is based on the activation of complement detected with an antibody against activated C9. The specificity of the three pathways was assessed using purified human MBL and mouse monoclonal antibodies against human C1q and pig factor D. Sera from 103 pigs and 38 newborn pigs were analyzed. Finally, functional activity of all pathways was assessed in vitro and in vivo in the absence and presence of complement inhibitors.

Results: The detection antibody showed cross-reactivity against pig. Normal pig serum showed activity in all pathways however about 10-fold more serum was required to obtain values comparable to human serum. Anti-human C1q and anti-pig factor D antibodies abolished classical and alternative pathway activity, respectively. Sera with low lectin pathway activity reconstituted with purified human MBL, fully recovered this activity. No deficiencies were found in classical or alternative pathway, whereas the lectin pathway showed reduced activity in a substantial number of pigs, similar to the situation in humans. Finally, the assay was successfully used to evaluate and monitor inhibition of pig complement in vitro and in vivo.

Conclusions: The human complement Wielisa test can be used for functional evaluation of all complement pathways in pig serum.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn / blood
  • Animals, Newborn / immunology
  • Complement Activation / physiology*
  • Complement System Proteins / immunology
  • Complement System Proteins / physiology
  • Feasibility Studies
  • Humans
  • Immunologic Tests / methods*
  • Signal Transduction / immunology
  • Species Specificity
  • Swine / blood
  • Swine / immunology*

Substances

  • Complement System Proteins