Skip to main content
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1977 Aug;60(2):465–472. doi: 10.1172/JCI108797

The Production of Antibody against Human Leukocytic Pyrogen

Charles A Dinarello 1, Lois Renfer 1, Sheldon M Wolff 1
PMCID: PMC372389  PMID: 559692

Abstract

Human peripheral blood leukocytes were stimulated with killed staphylococci in vitro to release leukocytic pyrogen (LP). Supernates from these stimulated leukocytes were concentrated, emulsified in Freund's complete adjuvant, and injected intradermally into rabbits. After seven monthly booster injections, rabbit antiserum destroyed the pyrogenic activity of human LP, and the titer of this neutralizing ability increased in the subsequent 7 mo. The pyrogen-neutralizing capacity of the rabbit antiserum was recovered in the globulin fraction, the IgG and IgM peaks of Sephadex G-200, and the acid-eluted fraction of a goat anti-rabbit IgG immunoadsorbant. The neutralizing antibody was specific for human LP inasmuch as it had no effect on rabbit, guinea pig, or monkey LP. When coupled to Sepharose, this antibody bound human LP; after acid elution from this immunoadsorbant, LP was recovered without loss of biologic or chemical characteristics. The antiserum was also absorbed with stimulated leukocyte supernates which did not contain LP, and this had no effect on the titer of anti-LP. Crude human LP, eluted from immunoadsorbant columns prepared from absorbed antiserum, contained significantly reduced contaminating protein when evaluated by polyacrylamide gel electrophoresis. These studies have established that specific antibody to human leukocytic pyrogen can be produced. This antibody is useful in the further study and purification of leukocytic pyrogen and its role in the pathogenesis of human fever.

Full text

PDF
465

Selected References

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

  1. Anfinsen C. B., Bose S., Corley L., Gurari-Rotman D. Partial purification of human interferon by affinity chromatography. Proc Natl Acad Sci U S A. 1974 Aug;71(8):3139–3142. doi: 10.1073/pnas.71.8.3139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Atkins E., Bodel P. Fever. N Engl J Med. 1972 Jan 6;286(1):27–34. doi: 10.1056/NEJM197201062860109. [DOI] [PubMed] [Google Scholar]
  3. Bodel P., Atkins E. Human leukocyte pyrogen producing fever in rabbits. Proc Soc Exp Biol Med. 1966 Mar;121(3):943–946. doi: 10.3181/00379727-121-30931. [DOI] [PubMed] [Google Scholar]
  4. Bodel P., Miller H. Pyrogen from mouse macrophages causes fever in mice. Proc Soc Exp Biol Med. 1976 Jan;151(1):93–96. doi: 10.3181/00379727-151-39150. [DOI] [PubMed] [Google Scholar]
  5. Böyum A. Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g. Scand J Clin Lab Invest Suppl. 1968;97:77–89. [PubMed] [Google Scholar]
  6. Cuatrecasas P., Wilchek M., Anfinsen C. B. Selective enzyme purification by affinity chromatography. Proc Natl Acad Sci U S A. 1968 Oct;61(2):636–643. doi: 10.1073/pnas.61.2.636. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DIXON M. A nomogram for ammonium sulphate solutions. Biochem J. 1953 Jun;54(3):457–458. doi: 10.1042/bj0540457. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dinarello C. A., Goldin N. P., Wolff S. M. Demonstration and characterization of two distinct human leukocytic pyrogens. J Exp Med. 1974 Jun 1;139(6):1369–1381. doi: 10.1084/jem.139.6.1369. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Greisman S. E., Hornick R. B. On the demonstration of circulating human endogenous pyrogen. Proc Soc Exp Biol Med. 1972 Feb;139(2):690–697. doi: 10.3181/00379727-139-36217. [DOI] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Mickenberg I. D., Root R. K., Wolff S. M. Bactericidal and metabolic properties of human eosinophils. Blood. 1972 Jan;39(1):67–80. [PubMed] [Google Scholar]
  12. Murphy P. A., Chesney P. J., Wood W. B., Jr Further purification of rabbit leukocyte pyrogen. J Lab Clin Med. 1974 Feb;83(2):310–322. [PubMed] [Google Scholar]
  13. Nordlund J. J., Root R. K., Wolff S. M. Studies on the origin of human leukocytic pyrogen. J Exp Med. 1970 Apr 1;131(4):727–743. doi: 10.1084/jem.131.4.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Perlow M., Dinarello C. A., Wolff S. M. A primate model for the study of human fever. J Infect Dis. 1975 Aug;132(2):157–164. doi: 10.1093/infdis/132.2.157. [DOI] [PubMed] [Google Scholar]
  15. Rawlins M. D., Cranston W. I., Luff R. H. Transferable pyrogen in human experimental fever. Clin Sci. 1971 Feb;40(2):193–196. doi: 10.1042/cs0400193. [DOI] [PubMed] [Google Scholar]
  16. Root R. K., Nordlund J. J., Wolff S. M. Factors affecting the quantitative production and assay of human leukocytic pyrogen. J Lab Clin Med. 1970 Apr;75(4):679–693. [PubMed] [Google Scholar]
  17. WOLFF S. M., MULHOLLAND J. H., WARD S. B. QUANTITATIVE ASPECTS OF THE PYROGENIC RESPONSE OF RABBITS TO ENDOTOXIN. J Lab Clin Med. 1965 Feb;65:268–275. [PubMed] [Google Scholar]
  18. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES