Skip to main content
PMC home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1980 May;40(2):283–291.

Correlation of disease activity and Clq-binding immune complexes with the neutrophil inclusions which form in the presence of SLE sera

E R Hurd, H E Jasin, J N Gilliam
PMCID: PMC1536984  PMID: 6254704

Abstract

Intracytoplasmic inclusions containing immunoglobulin (Ig) and complement (C3) are found in normal neutrophils (PMN) after incubation with sera from patients with SLE. These inclusions are believed to be immune complexes removed by phagocytosis from the SLE patients' sera in vitro. Similar inclusions were also noted in the circulating PMN from some patients with SLE. In the present study we have examined the relationship between the presence of intracytoplasmic inclusions and various clinical and laboratory features of SLE. Blood from forty-five patients with SLE was drawn and separated at 37°C. Fresh heparinized blood was also obtained from normal volunteers and allowed to stand for 90 min at 37°C. The buffy coat cells from both normal and SLE groups were removed, centrifuged, washed and examined (direct method) or incubated in the SLE sera for 90 min at 37°C (indirect method). Slides of washed cells were prepared in the cytocentrifuge, stained with fluorescein-conjugated goat anti-human IgG, IgM, IgA and C3 and examined under ultra-violet light.

By the direct method, 24% of patients had small intracytoplasmic inclusions in their neutrophils when stained for IgG suggesting that immune complexes were phagocytosed in vivo. None of twenty-one normal controls had similar inclusions. By the indirect method, 62% of SLE patients were positive for IgG, 15% for IgM, 8% for IgA and 31% for C3. None of the twelve normal controls were positive.

By the indirect method, PMN inclusions containing both IgG and IgM correlated with clinical activity (P<0·001), depressed serum complement (CH50, P=0·026; and C3, P<0·051), cryoglobulinaemia (P=0·014), anti-nDNA antibodies (P<0·001) and Clq-binding immune complexes (P=0·008). A suggestive correlation with granulocytopenia was also observed. The presence of inclusions containing IgG alone did not correlate with any of these parameters. C3 and IgM appeared to be mutually exclusive, i.e. neither was present simultaneously. These findings suggest (1) that normal PMN on exposure to SLE sera develop intracytoplasmic inclusions by phagocytosis of immune complexes, (2) the presence of such complexes correlates with a number of parameters of disease activity, particularly when IgG and IgM are both present and (3) such complexes may be phagocytosed in vivo as suggested by the presence of inclusions in vivo and contribute to a number of granulocyte disturbances seen in patients with SLE. These abnormalities in granulocyte function may be important, predisposing factors for infection in patients with active SLE.

Full text

PDF
283

Selected References

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

  1. Arana R., Seligmann M. Antibodies to native and denatured deoxyribonucleic acid in systemic lupus erythematosus. J Clin Invest. 1967 Nov;46(11):1867–1882. doi: 10.1172/JCI105677. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bardana E. J., Jr, Harbeck R. J., Hoffman A. A., Pirofsky B., Carr R. I. The prognostic and therapeutic implications of DNA:anti-DNA immune complexes in systemic lupus erythematosus (SLE). Am J Med. 1975 Oct;59(4):515–522. doi: 10.1016/0002-9343(75)90259-4. [DOI] [PubMed] [Google Scholar]
  3. Brandt L., Hedberg H. Impaired phagocytosis by peripheral blood granulocytes in systemic lupus erythematosus. Scand J Haematol. 1969;6(5):348–353. doi: 10.1111/j.1600-0609.1969.tb02420.x. [DOI] [PubMed] [Google Scholar]
  4. Brouet J. C., Clauvel J. P., Danon F., Klein M., Seligmann M. Biologic and clinical significance of cryoglobulins. A report of 86 cases. Am J Med. 1974 Nov;57(5):775–788. doi: 10.1016/0002-9343(74)90852-3. [DOI] [PubMed] [Google Scholar]
  5. CHRISTIAN C. L., HATFIELD W. B., CHASE P. H. Systemic lupus erythematosus. Cryoprecipitation of sera. J Clin Invest. 1963 Jun;42:823–829. doi: 10.1172/JCI104774. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cats A., Lafeber G. J., Klein F. Immunoglobulin phagocytosis by granulocytes from sera and synovial fluids in various rheumatoid and nonrheumatoid diseases. Ann Rheum Dis. 1975 Apr;34(2):146–155. doi: 10.1136/ard.34.2.146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Duckham D. J., Rhyne R. L., Jr, Smith F. E., Williams R. C., Jr Retardation of colony growth of in vitro bone marrow culture using sera from patients with Felty's syndrome, disseminated lupus erythematosus (SLE), rheumatoid arthritis, and other disease states. Arthritis Rheum. 1975 Jul-Aug;18(4):323–333. doi: 10.1002/art.1780180405. [DOI] [PubMed] [Google Scholar]
  8. Ginsberg B., Keiser H. A Millipore filter assay for antibodies to native DNA in sera of patients with systemic lupus erythematosus. Arthritis Rheum. 1973 Mar-Apr;16(2):199–207. doi: 10.1002/art.1780160210. [DOI] [PubMed] [Google Scholar]
  9. Grey H. M., Kohler P. F. Cryoimmunoglobulins. Semin Hematol. 1973 Apr;10(2):87–112. [PubMed] [Google Scholar]
  10. Hanauer L. B., Christian C. L. Studies of cryoproteins in systemic lupus erythematosus. J Clin Invest. 1967 Mar;46(3):400–408. doi: 10.1172/JCI105541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. JESSAR R. A., LAMONT-HAVERS R. W., RAGAN C. Natural history of lupus erythematosus disseminatus. Ann Intern Med. 1953 Apr;38(4):717–731. doi: 10.7326/0003-4819-38-4-717. [DOI] [PubMed] [Google Scholar]
  12. Jasin H. E., Orozco J. H., Ziff M. Serum heat-labile opsonins in systemic lupus erythematosus. J Clin Invest. 1974 Feb;53(2):343–353. doi: 10.1172/JCI107566. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Johnson G. D., Edmonds J. P., Holborow E. J. Precipitating antibody to D.N.A. detected by two-stage electroimmunodiffusion. Study in S.L.E. and in rheumatoid arthritis. Lancet. 1973 Oct 20;2(7834):883–885. doi: 10.1016/s0140-6736(73)92008-4. [DOI] [PubMed] [Google Scholar]
  14. LERMAN L. S. Structural considerations in the interaction of DNA and acridines. J Mol Biol. 1961 Feb;3:18–30. doi: 10.1016/s0022-2836(61)80004-1. [DOI] [PubMed] [Google Scholar]
  15. Landry M. Phagocyte function and cell-mediated immunity in systemic lupus erythematosus. Arch Dermatol. 1977 Feb;113(2):147–154. [PubMed] [Google Scholar]
  16. MICHAEL S. R., VURAL I. L., BASSEN F. A., SCHAEFER L. The hematologic aspects of disseminated (systemic) lupus erythematosus. Blood. 1951 Nov;6(11):1059–1072. [PubMed] [Google Scholar]
  17. McPhaul J. J., Jr Cryoimmunoglobulinaemia in patients with primary renal disease and systemic lupus erythematosus. I. IgG- and DNA-binding assessed by co-precipitation. Clin Exp Immunol. 1978 Jan;31(1):131–140. [PMC free article] [PubMed] [Google Scholar]
  18. Meltzer M., Franklin E. C. Cryoglobulinemia--a study of twenty-nine patients. I. IgG and IgM cryoglobulins and factors affecting cryoprecipitability. Am J Med. 1966 Jun;40(6):828–836. doi: 10.1016/0002-9343(66)90199-9. [DOI] [PubMed] [Google Scholar]
  19. Mowat A. G., Baum J. Chemotaxis of polymorphonuclear leukocytes from patients with rheumatoid arthritis. J Clin Invest. 1971 Dec;50(12):2541–2549. doi: 10.1172/JCI106754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Nelson R. A., Jr, Jensen J., Gigli I., Tamura N. Methods for the separation, purification and measurement of nine components of hemolytic complement in guinea-pig serum. Immunochemistry. 1966 Mar;3(2):111–135. doi: 10.1016/0019-2791(66)90292-8. [DOI] [PubMed] [Google Scholar]
  21. Paoletti C., LePecq J. B., Lehman I. R. The use of ethidium bromide-circular DNA complexes for the fluorometric analysis of breakage and joining of DNA. J Mol Biol. 1971 Jan 14;55(1):75–100. doi: 10.1016/0022-2836(71)90282-8. [DOI] [PubMed] [Google Scholar]
  22. Pincus T., Schur P. H., Rose J. A., Decker J. L., Talal N. Measurement of serum DNA-binding activity in systemic lupus erythematosus. N Engl J Med. 1969 Sep 25;281(13):701–705. doi: 10.1056/NEJM196909252811304. [DOI] [PubMed] [Google Scholar]
  23. Ploem J. S. A study of filters and light sources in immunofluorescence microscopy. Ann N Y Acad Sci. 1971 Jun 21;177:414–429. doi: 10.1111/j.1749-6632.1971.tb35070.x. [DOI] [PubMed] [Google Scholar]
  24. ROPES M. W. OBSERVATIONS ON THE NATURAL COURSE OF DISSEMINATED LUPUS ERYTHEMATOSUS. Medicine (Baltimore) 1964 May;43:387–391. doi: 10.1097/00005792-196405000-00016. [DOI] [PubMed] [Google Scholar]
  25. Robitaille P., Tan E. M. Relationship between deoxyribonucleoprotein and deoxyribonucleic acid antibodies in systemic lupus erythematosus. J Clin Invest. 1973 Feb;52(2):316–323. doi: 10.1172/JCI107187. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schur P. H., Sandson J. Immunologic factors and clinical activity in systemic lupus erythematosus. N Engl J Med. 1968 Mar 7;278(10):533–538. doi: 10.1056/NEJM196803072781004. [DOI] [PubMed] [Google Scholar]
  27. Sontheimer R. D., Gilliam J. N. A reappraisal of the relationship between subepidermal immunoglobulin deposits and DNA antibodies in systemic lupus erythematosus: a study using the Crithidia luciliae immunofluorescence anti-DNA assay. J Invest Dermatol. 1979 Jan;72(1):29–32. doi: 10.1111/1523-1747.ep12530158. [DOI] [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. Taichman N. S., Pruzanski W., Ranadive N. S. Release of intracellular constituents from rabbit polymorphonuclear leukocytes exposed to soluble and insoluble immune complexes. Int Arch Allergy Appl Immunol. 1972;43(2):182–195. doi: 10.1159/000230836. [DOI] [PubMed] [Google Scholar]
  30. Tan E. M., Schur P. H., Carr R. I., Kunkel H. G. Deoxybonucleic acid (DNA) and antibodies to DNA in the serum of patients with systemic lupus erythematosus. J Clin Invest. 1966 Nov;45(11):1732–1740. doi: 10.1172/JCI105479. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Turner R. A., Schumacher R., Myers A. R. Phagocytic function of polymorphonuclear leukocytes in rheumatic diseases. J Clin Invest. 1973 Jul;52(7):1632–1635. doi: 10.1172/JCI107342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Vaughan J. H., Barrnett E. V., Sobel M. V., Jacox R. F. Intracytoplasmic inclusions of immunoglobulins in rheumatoid arthritis and other diseases. Arthritis Rheum. 1968 Apr;11(2):125–134. doi: 10.1002/art.1780110202. [DOI] [PubMed] [Google Scholar]
  33. Winfield J. B., Koffler D., Kunkel H. G. Specific concentration of polynucleotide immune complexes in the cryoprecipitates of patients with systemic lupus erythematosus. J Clin Invest. 1975 Sep;56(3):563–570. doi: 10.1172/JCI108125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zubler R. H., Lange G., Lambert P. H., Miescher P. A. Detection of immune complexes in unheated sera by modified 125I-Clq binding test. Effect of heating on the binding of Clq by immune complexes and application of the test to systemic lupus erythematosus. J Immunol. 1976 Jan;116(1):232–235. [PubMed] [Google Scholar]
  35. van Rooijen N. Immune complexes in the spleen. Replacement of immune complexes trapped in spleen follicles by new immune complexes from the circulation. Int Arch Allergy Appl Immunol. 1975;49(6):754–762. [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES