Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1980 Sep;66(3):457–464. doi: 10.1172/JCI109876

Inhibition of polymorphonuclear leukocyte chemiluminescence for detection of immune complexes in human sera.

N J Doll, M R Wilson, J E Salvaggio
PMCID: PMC371673  PMID: 6156953

Abstract

An assay for the detection and quantitation of immune complexes is described. Experimental immune complexes or aggregated human gamma globulin (AHG) were incubated with polymorphonuclear leukocytes (PMN). After challenge of the PMN with opsonized zymosan, chemiluminescence was recorded in a scintillation spectrometer. A quantitative inhibition of chemiluminescence could be demonstrated by the interaction of PMN with immune complexes or AHG. Experimental immune complexes of bovine serum albumin-anti-bovine serum albumin were formed and tested by this assay, and immune complexes formed near antigen excess were best described by this technique. The technique was used to demonstrate immune complexes in the sera from patients with systemic lupus erythematosus, rheumatoid arthritis, and vasculitis. Immune complexes were quantitated by reference to a standard curve using AHG. By this technique, normal human sera had < 10 micrograms of AHG per milliliter of serum. Immune complexes at levels above this were detected in 9/15 patients with systemic lupus erythematosus, 18/30 patients with rheumatoid arthritis, and 2/5 patients with vasculitis. Therefore, this assay is a sensitive, simple method for measurement of circulating immune complexes in the sera of patients with certain connective tissue diseases.

Full text

PDF
457

Selected References

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

  1. Agnello V., Koffler D., Eisenberg J. W., Winchester R. J., Kundel H. G. C1g precipitins in the sera of patients with systemic lupus erythematosus and other hypocomplementemic states: characterization of high and low molecular weight types. J Exp Med. 1971 Sep 1;134(3 Pt 2):228s–241s. [PubMed] [Google Scholar]
  2. Agnello V., Winchester R. J., Kunkel H. G. Precipitin reactions of the C1q component of complement with aggregated gamma-globulin and immune complexes in gel diffusion. Immunology. 1970 Dec;19(6):909–919. [PMC free article] [PubMed] [Google Scholar]
  3. Allen R. C. Evaluation of serum opsonic capacity by quantitating the initial chemiluminescent response from phagocytizing polymorphonuclear leukocytes. Infect Immun. 1977 Mar;15(3):828–833. doi: 10.1128/iai.15.3.828-833.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Allen R. C., Stjernholm R. L., Reed M. A., Harper T. B., 3rd, Gupta S., Steele R. H., Waring W. W. Correlation of metabolic and chemiluminescent responses of granulocytes from three female siblings with chronic granulomatous disease. J Infect Dis. 1977 Oct;136(4):510–518. doi: 10.1093/infdis/136.4.510. [DOI] [PubMed] [Google Scholar]
  5. CHRISTIAN C. L. Studies of aggregated gamma-globulin. I. Sedimentation, electrophoretic and anticomplementary properties. J Immunol. 1960 Jan;84:112–116. [PubMed] [Google Scholar]
  6. Camussi G., Tetta C., Cappio F. C. Detection of immune complexes on the surface of polymorphonuclear neutrophils. Int Arch Allergy Appl Immunol. 1979;58(2):135–139. doi: 10.1159/000232185. [DOI] [PubMed] [Google Scholar]
  7. Chia D., Barnett E. V., Yamagata J., Knutson D., Restivo C., Furst D. Quantitation and characterization of soluble immune complexes precipitated from sera by polyethylene glycol (PEG). Clin Exp Immunol. 1979 Sep;37(3):399–407. [PMC free article] [PubMed] [Google Scholar]
  8. Faber V., Elling P. Leucocyte-specific anti-nuclear factors in patients with felty's syndrome, rheumatoid arthritis, systemic lupus erythematosus and other diseases. Acta Med Scand. 1966 Mar;179(3):257–267. doi: 10.1111/j.0954-6820.1966.tb05457.x. [DOI] [PubMed] [Google Scholar]
  9. Goldstein I. M., Feit F., Weissmann G. Enhancement of nitroblue tetrazolium dye reduction by leukocytes exposed to a component of complement in the absence of phagocytosis. J Immunol. 1975 Jan;114(1 Pt 2):516–518. [PubMed] [Google Scholar]
  10. Goldstein I. M., Kaplan H. B., Radin A., Frosch M. Independent effects of IgG and complement upon human polymorphonuclear leukocyte function. J Immunol. 1976 Oct;117(4):1282–1287. [PubMed] [Google Scholar]
  11. Gupta R. C., McDuffie F. C., Huston K. A., Tappeiner G., Meurer M., Jordon R. E., Luthra H. S., Hunder G. G., Ilstrup D. Comparison of three immunoassays for immune complexes in rheumatoid arthritis. Arthritis Rheum. 1979 May;22(5):433–439. doi: 10.1002/art.1780220501. [DOI] [PubMed] [Google Scholar]
  12. Gupta R. C., McDuffie F. C., Tappeiner G., Jordon R. E. Binding of soluble immune complexes to Raji lymphocytes. Role of receptors for complement components, C1q and C3-C3b. Immunology. 1978 Apr;34(4):751–761. [PMC free article] [PubMed] [Google Scholar]
  13. Henson P. M. Interaction of cells with immune complexes: adherence, release of constituents, and tissue injury. J Exp Med. 1971 Sep 1;134(3 Pt 2):114s–135s. [PubMed] [Google Scholar]
  14. Henson P. M., Johnson H. B., Spiegelberg H. L. The release of granule enzymes from human neutrophils stimulated by aggregated immunoglobulins of different classes and subclasses. J Immunol. 1972 Dec;109(6):1182–1192. [PubMed] [Google Scholar]
  15. Henson P. M., Oades Z. G. Stimulation of human neutrophils by soluble and insoluble immunoglobulin aggregates. Secretion of granule constituents and increased oxidation of glucose. J Clin Invest. 1975 Oct;56(4):1053–1061. doi: 10.1172/JCI108152. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Henson P. M. The immunologic release of constituents from neutrophil leukocytes. II. Mechanisms of release during phagocytosis, and adherence to nonphagocytosable surfaces. J Immunol. 1971 Dec;107(6):1547–1557. [PubMed] [Google Scholar]
  17. Johnston R. B., Jr, Lehmeyer J. E. Elaboration of toxic oxygen by-products by neutrophils in a model of immune complex disease. J Clin Invest. 1976 Apr;57(4):836–841. doi: 10.1172/JCI108359. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Luthra H. S., McDuffie F. C., Hunder G. G., Samayoa E. A. Immune complexes in sera and synovial fluids of patients with rheumatoid arthritis. Radioimmunoassay with monocylonal rheumatoid factor. J Clin Invest. 1975 Aug;56(2):458–466. doi: 10.1172/JCI108112. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mackel S. E., Tappeiner G., Brumfield H., Jordan R. E. Circulating immune complexes in cutaneous vasculitis. Detection with C1q and monoclonal rheumatoid factor. J Clin Invest. 1979 Dec;64(6):1652–1660. doi: 10.1172/JCI109627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mantovani B. Different roles of IgG and complement receptors in phagocytosis by polymorphonuclear leukocytes. J Immunol. 1975 Jul;115(1):15–17. [PubMed] [Google Scholar]
  21. Matheisz J. S., Allen J. C. Fatigue of human polymorphonuclear leukocyte function associated with bacterial ingestion. Clin Immunol Immunopathol. 1979 Feb;12(2):125–131. doi: 10.1016/0090-1229(79)90001-1. [DOI] [PubMed] [Google Scholar]
  22. McDougal J. S., Redecha P. B., Inman R. D., Christian C. L. Binding of immunoglobulin G aggregates and immune complexes in human sera to Staphylococci containing protein A. J Clin Invest. 1979 Apr;63(4):627–636. doi: 10.1172/JCI109345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Newman S. L., Johnston R. B., Jr Role of binding through C3b and IgG in polymorphonuclear neutrophil function: studies with trypsin-generated C3b. J Immunol. 1979 Oct;123(4):1839–1846. [PubMed] [Google Scholar]
  24. Nishimaki T., Kano K., Milgrom F. Studies on immune complexes in rheumatoid arthritis. Arthritis Rheum. 1978 Jul-Aug;21(6):639–644. doi: 10.1002/art.1780210605. [DOI] [PubMed] [Google Scholar]
  25. Nydegger U. E., Lambert P. H., Gerber H., Miescher P. A. Circulating immune complexes in the serum in systemic lupus erythematosus and in carriers of hepatitis B antigen. Quantitation by binding to radiolabeled C1q. J Clin Invest. 1974 Aug;54(2):297–309. doi: 10.1172/JCI107765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schocket A. L., Kohler P. F. Lymphocytotoxic antibodies in systemic lupus erythematosus and clinically related diseases. Arthritis Rheum. 1979 Oct;22(10):1060–1063. doi: 10.1002/art.1780221003. [DOI] [PubMed] [Google Scholar]
  27. Scribner D. J., Fahrney D. Neutrophil receptors for IgG and complement: their roles in the attachment and ingestion phases of phagocytosis. J Immunol. 1976 Apr;116(4):892–897. [PubMed] [Google Scholar]
  28. Steffelaar J. W., Ten Kate F. J., Nap M., Swaak A. J., De Graaffreitsma C. B., Van Elven E. H., Feltkamp-Vroom T. M. Immune complex detection by immunofluorescence on polymorphonuclear leucocytes. Clin Exp Immunol. 1977 Mar;27(3):391–396. [PMC free article] [PubMed] [Google Scholar]
  29. Theofilopoulos A. N., Wilson C. B., Dixon F. J. The Raji cell radioimmune assay for detecting immune complexes in human sera. J Clin Invest. 1976 Jan;57(1):169–182. doi: 10.1172/JCI108257. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. UCLA conference. Circulating immune complexes: their immunochemistry, detection, and importance. Ann Intern Med. 1979 Sep;91(3):430–440. doi: 10.7326/0003-4819-91-3-430. [DOI] [PubMed] [Google Scholar]
  31. Weissmann G., Zurier R. B., Spieler P. J., Goldstein I. M. Mechanisms of lysosomal enzyme release from leukocytes exposed to immune complexes and other particles. J Exp Med. 1971 Sep 1;134(3 Pt 2):149s–165s. [PubMed] [Google Scholar]
  32. Winchester R. J., Kunkel H. G., Agnello V. Occurrence of -globulin complexes in serum and joint fluid of rheumatoid arthritis patients: use of monoclonal rheumatoid factors as reagents for their demonstration. J Exp Med. 1971 Sep 1;134(3 Pt 2):286s–295s. [PubMed] [Google Scholar]

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

RESOURCES