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. 1985 Aug;61(2):224–231.

The role of metabolism in the immunogenicity of drugs: production of antibodies to a horseradish peroxidase generated conjugate paracetamol.

J Chesham, G E Davies
PMCID: PMC1577289  PMID: 2412741

Abstract

The allergic response to small chemically inert molecules in thought to require their enzymatic conversion to reactive metabolites which are then endowed with the capacity to bind covalently to host proteins and produce immunogenic hapten-carrier conjugates. In contrast to previous studies in which hapten-carrier conjugates have been generated following chemical modification of drugs we have examined the immunogenicity of paracetamol following direct conjugation to carrier proteins with horseradish peroxidase (HRP). Highly substituted conjugates of paracetamol with keyhole limpet haemocyanin (KLH) or bovine serum albumin (BSA) were generated using HRP. The KLH conjugate was used to immunize Balb/C mice. IgM and IgG (predominantly IgG1) responses were observed and shown by enzyme-linked immunosorbent assay (ELISA) to be hapten-specific. Manipulations of HRP levels permitted substitution of KLH to varying extents with paracetamol. Such conjugates were tested for their ability to induce a hapten-specific immune response. It was determined that substitution of 1 mol of KLH with 700 mol of paracetamol was sufficient to generate an anti-hapten response. These data suggest a mechanism by which protein-non-reactive drugs may be rendered immunogenic and provide a method for demonstrating the presence of serum antibodies reactive with drug metabolites.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. CARR E. A., Jr Drug allergy. Pharmacol Rev. 1954 Dec;6(4):365–424. [PubMed] [Google Scholar]
  2. Drayer D. E. Pharmacologically active metabolites of drugs and other foreign compounds. Clinical, pharmacological, therapeutic and toxicological considerations. Drugs. 1982 Dec;24(6):519–542. doi: 10.2165/00003495-198224060-00003. [DOI] [PubMed] [Google Scholar]
  3. Eisner E. V., Shahidi N. T. Immune thrombocytopenia due to a drug metabolite. N Engl J Med. 1972 Aug 24;287(8):376–381. doi: 10.1056/NEJM197208242870803. [DOI] [PubMed] [Google Scholar]
  4. FARR R. S. A quantitative immunochemical measure of the primary interaction between I BSA and antibody. J Infect Dis. 1958 Nov-Dec;103(3):239–262. doi: 10.1093/infdis/103.3.239. [DOI] [PubMed] [Google Scholar]
  5. Gelboin H. V. A microsome-dependent binding of benzo[a]pyrene to DNA. Cancer Res. 1969 Jun;29(6):1272–1276. [PubMed] [Google Scholar]
  6. Green T., Hathway D. E. Interactions of vinyl chloride with rat-liver DNA in vivo. Chem Biol Interact. 1978 Sep;22(2-3):211–224. doi: 10.1016/0009-2797(78)90126-6. [DOI] [PubMed] [Google Scholar]
  7. Jollow D. J., Mitchell J. R., Potter W. Z., Davis D. C., Gillette J. R., Brodie B. B. Acetaminophen-induced hepatic necrosis. II. Role of covalent binding in vivo. J Pharmacol Exp Ther. 1973 Oct;187(1):195–202. [PubMed] [Google Scholar]
  8. Kappus H., Remmer H. Irreversible protein binding of [14-C]imipramine with rat and human liver microsomes. Biochem Pharmacol. 1975 May 15;24(10):1079–1084. doi: 10.1016/0006-2952(75)90193-8. [DOI] [PubMed] [Google Scholar]
  9. Levine B. B. Immunochemical mechanisms of drug allergy. Annu Rev Med. 1966;17:23–38. doi: 10.1146/annurev.me.17.020166.000323. [DOI] [PubMed] [Google Scholar]
  10. Mitchison N. A. The carrier effect in the secondary response to hapten-protein conjugates. II. Cellular cooperation. Eur J Immunol. 1971 Jan;1(1):18–27. doi: 10.1002/eji.1830010104. [DOI] [PubMed] [Google Scholar]
  11. Nelson S. D., Dahlin D. C., Rauckman E. J., Rosen G. M. Peroxidase-mediated formation of reactive metabolites of acetaminophen. Mol Pharmacol. 1981 Jul;20(1):195–199. [PubMed] [Google Scholar]
  12. Potter W. Z., Davis D. C., Mitchell J. R., Jollow D. J., Gillette J. R., Brodie B. B. Acetaminophen-induced hepatic necrosis. 3. Cytochrome P-450-mediated covalent binding in vitro. J Pharmacol Exp Ther. 1973 Oct;187(1):203–210. [PubMed] [Google Scholar]
  13. Rahimtula A. D., O'Brien P. J. Hydroperoxide catalyzed liver microsomal aromatic hydroxylation reactions involving cytochrome P-450. Biochem Biophys Res Commun. 1974 Sep 9;60(1):440–447. doi: 10.1016/0006-291x(74)90223-x. [DOI] [PubMed] [Google Scholar]

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