Skip to main content

Some NLM-NCBI services and products are experiencing heavy traffic, which may affect performance and availability. We apologize for the inconvenience and appreciate your patience. For assistance, please contact our Help Desk at info@ncbi.nlm.nih.gov.

The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1976 Jan;57(1):169–182. doi: 10.1172/JCI108257

The Raji cell radioimmune assay for detecting immune complexes in human sera.

A N Theofilopoulos, C B Wilson, F J Dixon
PMCID: PMC436637  PMID: 128562

Abstract

A sensitivie and simple procedure for the detection and quantitation of soluble complement (C)- fixing immune complexes in sera of patients with various disease states has been developed by utilizing C receptors on Raji cells. These cells lack membrane-bound immunoglobulin but have receptors for IgG Fc, C3b, C3d, and possibly with other C proteins. Uptake experiments showed that both aggregated human gamma globulin (AHG) and 7S IgG bound to receptors for IgG Fc; however, AHG reacted with C bound to cells only via receptors for C and this binding was much more efficient than via IgG Fc receptors. AHG was used as an in vitro model of human immune complexes and its uptake by Raji cells was quantitated by 125I-radiolabeled antihuman IgG. The limit of sensitivity of this test was 6 mug AHG/ml serum. The ability of Raji cells to detect AHG in serum depended on the amount of radioactive antibody used and the size of aggregates. The presence of an excess of C somewhat inhibited binding of AHG containing C to Raji cells. The efficient binding of AHG by receptors for C on Raji cells was used for the detection and quantitation of immune complexes in human sera. Raji cells were incubated with sera to be tested and then reacted with excess radiolabeled antihuman IgG; the amount of radioactivity bound to the washed cells was determined and referred to a standard curve of radioactive antibody uptake by cells previously incubated with increasing amounts of AHG in serum. Thereby immune complexes were detected and quantitated in serum hepatitis, systemic lupus erythematosus, vasculitis, subacute sclerosing panencephalitis, dengue hemorrhagic fever, and malignancies.

Full text

PDF
169

Selected References

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

  1. 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]
  2. Ahmed A., Strong D. M., Sell K. W., Thurman G. B., Knudsen R. C., Wistar R., Jr, Grace W. R. Demonstration of a blocking factor in the plasma and spinal fluid of patients with subacute sclerosing panencephalitis. I. Partial characterization. J Exp Med. 1974 Apr 1;139(4):902–924. doi: 10.1084/jem.139.4.902. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Ambrose K. R., Anderson N. G., Coggin J. H., Jr Cytostatic antibody and SV40 tumour immunity in hamsters. Nature. 1971 Oct 1;233(5318):321–324. doi: 10.1038/233321a0. [DOI] [PubMed] [Google Scholar]
  4. Baldwin R. W., Price M. R., Robins R. A. Blocking of lymphocyte-mediated cytotoxicity for rat hepatoma cells by tumour-specific antigen-antibody complexes. Nat New Biol. 1972 Aug 9;238(84):185–186. doi: 10.1038/newbio238185a0. [DOI] [PubMed] [Google Scholar]
  5. Bokisch V. A., Müller-Eberhard H. J., Cochrane C. G. Isolation of a fragment (C3a) of the third component of human complement containing anaphylatoxin and chemotactic activity and description of an anaphylatoxin inactivator of human serum. J Exp Med. 1969 May 1;129(5):1109–1130. doi: 10.1084/jem.129.5.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. CHRISTIAN C. L. Studies of aggregated gamma-globulin. I. Sedimentation, electrophoretic and anticomplementary properties. J Immunol. 1960 Jan;84:112–116. [PubMed] [Google Scholar]
  8. CLARK H. F., SHEPARD C. C. A DIALYSIS TECHNIQUE FOR PREPARING FLUORESCENT ANTIBODY. Virology. 1963 Aug;20:642–644. doi: 10.1016/0042-6822(63)90292-7. [DOI] [PubMed] [Google Scholar]
  9. Cochrane C. G., Koffler D. Immune complex disease in experimental animals and man. Adv Immunol. 1973;16(0):185–264. doi: 10.1016/s0065-2776(08)60298-9. [DOI] [PubMed] [Google Scholar]
  10. Costanza M. E., Pinn V., Schwartz R. S., Nathanson L. Carcinoembryonic antigen-antibody complexes in a patient with colonic carcinoma and nephrotic syndrome. N Engl J Med. 1973 Sep 6;289(10):520–522. doi: 10.1056/NEJM197309062891007. [DOI] [PubMed] [Google Scholar]
  11. Creighton W. D., Lambert P. H., Miescher P. A. Detection of antibodies and soluble antigen-antibody complexes by precipitation with polyethylene glycol. J Immunol. 1973 Oct;111(4):1219–1227. [PubMed] [Google Scholar]
  12. DIXON F. J., FELDMAN J. D., VAZQUEZ J. J. Experimental glomerulonephritis. The pathogenesis of a laboratory model resembling the spectrum of human glomerulonephritis. J Exp Med. 1961 May 1;113:899–920. doi: 10.1084/jem.113.5.899. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Dickler H. B., Kunkel H. G. Interaction of aggregated -globulin with B lymphocytes. J Exp Med. 1972 Jul 1;136(1):191–196. doi: 10.1084/jem.136.1.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Dickler H. B. Studies of the human lymphocyte receptor for heat-aggregated or antigen-complexed immunoglobulin. J Exp Med. 1974 Aug 1;140(2):508–522. doi: 10.1084/jem.140.2.508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Dixon F. J., Cochrane C. G. The pathogenicity of antigen-antibody complexes. Pathol Annu. 1970;5:355–379. [PubMed] [Google Scholar]
  16. Dixon F. J. Immune complex diseases. J Invest Dermatol. 1972 Dec;59(6):413–415. doi: 10.1111/1523-1747.ep12627589. [DOI] [PubMed] [Google Scholar]
  17. Eden A., Bianco C., Nussenzweig Mechanism of binding of soluble immune complexes to lymphocytes. Cell Immunol. 1973 Jun;7(3):459–473. doi: 10.1016/0008-8749(73)90210-4. [DOI] [PubMed] [Google Scholar]
  18. FERRARI A. Nitrogen determination by a continuous digestion and analysis system. Ann N Y Acad Sci. 1960 Jul 22;87:792–800. doi: 10.1111/j.1749-6632.1960.tb23236.x. [DOI] [PubMed] [Google Scholar]
  19. FRANKLIN E. C., HOLMAN H. R., MULLER-EBERHARD H. J., KUNKEL H. G. An unusual protein component of high molecular weight in the serum of certain patients with rheumatoid arthritis. J Exp Med. 1957 May 1;105(5):425–438. doi: 10.1084/jem.105.5.425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Friou G. J. Antinuclear antibodies: diagnostic significance and methods. Arthritis Rheum. 1967 Apr;10(2):151–159. doi: 10.1002/art.1780100210. [DOI] [PubMed] [Google Scholar]
  21. Gocke D. J., Howe C. Rapid detection of Australia antigen by counterimmunoelectrophoresis. J Immunol. 1970 Apr;104(4):1031–1034. [PubMed] [Google Scholar]
  22. Gocke D. J., Hsu K., Morgan C., Bombardieri S., Lockshin M., Christian C. L. Association between polyarteritis and Australia antigen. Lancet. 1970 Dec 5;2(7684):1149–1153. doi: 10.1016/s0140-6736(70)90339-9. [DOI] [PubMed] [Google Scholar]
  23. Gocke D. J., Hsu K., Morgan C., Bombardieri S., Lockshin M., Christian C. L. Vasculitis in association with Australia antigen. J Exp Med. 1971 Sep 1;134(3 Pt 2):330s–336s. [PubMed] [Google Scholar]
  24. Heimer R., Abruzzo J. L. A latex test for the detection of human IgG aggregates and IgG anti-IgG antibody. Immunochemistry. 1972 Sep;9(9):921–931. doi: 10.1016/0019-2791(72)90165-6. [DOI] [PubMed] [Google Scholar]
  25. Hellström I., Hellström K. E., Sjögren H. O., Warner G. A. Serum factors in tumor-free patients cancelling the blocking of cell-mediated tumor immunity. Int J Cancer. 1971 Sep 15;8(2):185–191. doi: 10.1002/ijc.2910080202. [DOI] [PubMed] [Google Scholar]
  26. ISHIZAKA K., ISHIZAKA T., BANOVITZ J. BIOLOGICAL ACTIVITIES OF AGGREGATED GAMMA-GLOBULIN. VII. MINIMUM SIZE OF AGGREGATED GAMMA-GLOBULIN OR ITS PIECE 3 REQUIRED FOR THE INDUCTION OF SKIN REACTIVITY AND COMPLEMENT FIXATION. J Immunol. 1965 Jun;94:825–832. [PubMed] [Google Scholar]
  27. Jewell D. P., MacLennan I. C. Circulating immune complexes in inflammatory bowel disease. Clin Exp Immunol. 1973 Jun;14(2):219–226. [PMC free article] [PubMed] [Google Scholar]
  28. KUNKEL H. G., TAN E. M. AUTOANTIBODIES AND DISEASE. Adv Immunol. 1964;27:351–395. doi: 10.1016/s0065-2776(08)60711-7. [DOI] [PubMed] [Google Scholar]
  29. Koffler D., Agnello V., Thoburn R., Kunkel H. G. Systemic lupus erythematosus: prototype of immune complex nephritis in man. J Exp Med. 1971 Sep 1;134(3 Pt 2):169s–179s. [PubMed] [Google Scholar]
  30. Koffler D., Carr R., Agnello V., Thoburn R., Kunkel H. G. Antibodies to polynucleotides in human sera: antigenic specificity and relation to disease. J Exp Med. 1971 Jul 1;134(1):294–312. doi: 10.1084/jem.134.1.294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Koffler D., Schur P. H., Kunkel H. G. Immunological studies concerning the nephritis of systemic lupus erythematosus. J Exp Med. 1967 Oct 1;126(4):607–624. doi: 10.1084/jem.126.4.607. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Kohler P. F., Cronin R. E., Hammond W. S., Olin D., Carr R. I. Chronic membranous glomerulonephritis caused by hepatitis B antigen-antibody immune complexes. Ann Intern Med. 1974 Oct;81(4):448–451. doi: 10.7326/0003-4819-81-4-448. [DOI] [PubMed] [Google Scholar]
  33. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  34. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  35. Mowbray J. F., Hoffbrand A. V., Holborow E. J., Seah P. P., Fry L. Circulating immune complexes in dermatitis herpetiformis. Lancet. 1973 Feb 24;1(7800):400–402. doi: 10.1016/s0140-6736(73)90253-5. [DOI] [PubMed] [Google Scholar]
  36. Myllylä G. Aggregation of human blood platelets by immune complexes in the sedimentation pattern test. Scand J Haematol Suppl. 1973;19:1–56. [PubMed] [Google Scholar]
  37. Notkins A. L., Mahar S., Scheele C., Goffman J. Infectious virus-antibody complex in the blood of chronically infected mice. J Exp Med. 1966 Jul 1;124(1):81–97. doi: 10.1084/jem.124.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. Oldstone M. B., Cooper N. R., Larson D. L. Formation and biologic role of polyoma virus-antibody complexes. A critical role for complement. J Exp Med. 1974 Aug 1;140(2):549–565. doi: 10.1084/jem.140.2.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Oldstone M. B., Dixon F. J. Pathogenesis of chronic disease associated with persistent lymphocytic choriomeningitis viral infection. I. Relationship of antibody production to disease in neonatally infected mice. J Exp Med. 1969 Mar 1;129(3):483–505. doi: 10.1084/jem.129.3.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Oldstone M. B. Immune complexes in cancer: demonstration of complexes in mice bearing neuroblastomas. J Natl Cancer Inst. 1975 Jan;54(1):223–228. doi: 10.1093/jnci/54.1.223. [DOI] [PubMed] [Google Scholar]
  42. Oldstone M. B., Theofilopoulos A. N., Gunvén P., Klein G. Immune complexes associated with neoplasia: presence of Epstein-Barr virus antigen-antibody complexes in Burkitt's lymphoma. Intervirology. 1974;4(5):292–302. doi: 10.1159/000149862. [DOI] [PubMed] [Google Scholar]
  43. Oldstone M. B. Virus neutralization and virus-induced immune complex disease. Virus-antibody union resulting in immunoprotection or immunologic injury--two sides of the same coin. Prog Med Virol. 1975;19:84–119. [PubMed] [Google Scholar]
  44. Onyewotu I. I., Holborow E. J., Johnson G. D. Detection and radioassay of soluble circulating immune complexes using guinea pig peritoneal exudate cells. Nature. 1974 Mar 8;248(5444):156–159. doi: 10.1038/248156a0. [DOI] [PubMed] [Google Scholar]
  45. Onyewotu I. I., Johnson P. M., Johnson G. D., Holborow E. J. Enhanced uptake by guinea-pig macrophages of radio-iodinated human aggregated immunoglobulin G in the presence of sera from rheumatoid patients with cutaneous vasculitis. Clin Exp Immunol. 1975 Feb;19(2):267–280. [PMC free article] [PubMed] [Google Scholar]
  46. PULVERTAFT J. V. A STUDY OF MALIGNANT TUMOURS IN NIGERIA BY SHORT-TERM TISSUE CULTURE. J Clin Pathol. 1965 May;18:261–273. doi: 10.1136/jcp.18.3.261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Pope R. M., Teller D. C., Mannik M. Intermediate complexes formed by self-association of IgG-rheumatoid factors. Ann N Y Acad Sci. 1975 Jun 13;256:82–87. doi: 10.1111/j.1749-6632.1975.tb36037.x. [DOI] [PubMed] [Google Scholar]
  48. Poskitt P. K., Poskitt T. R., Wallace J. H. Renal deposition of soluble immune complexes in mice bearing B-16 melanoma. Characterization of complexes and relationship to tumor progress. J Exp Med. 1974 Aug 1;140(2):410–425. doi: 10.1084/jem.140.2.410. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. 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]
  50. Ruddy S., Austen K. F. C3b inactivator of man. II. Fragments produced by C3b inactivator cleavage of cell-bound or fluid phase C3b. J Immunol. 1971 Sep;107(3):742–750. [PubMed] [Google Scholar]
  51. Sjögren H. O., Hellström I., Bansal S. C., Hellström K. E. Suggestive evidence that the "blocking antibodies" of tumor-bearing individuals may be antigen--antibody complexes. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1372–1375. doi: 10.1073/pnas.68.6.1372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Sobel A. T., Bokisch V. A., Müller-Eberhard H. J. C1q deviation test for the detection of immune complexes, aggregates of IgG, and bacterial products in human serum. J Exp Med. 1975 Jul 1;142(1):139–150. doi: 10.1084/jem.142.1.139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Sutherland J. C., Mardiney M. R., Jr Immune complex disease in the kidneys of lymphoma-leukemia patients: the presence of an oncornavirus-related antigen. J Natl Cancer Inst. 1973 Mar;50(3):633–644. doi: 10.1093/jnci/50.3.633. [DOI] [PubMed] [Google Scholar]
  54. 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]
  55. Theofilopoulos A. N., Bokisch V. A., Dixon F. J. Receptor for soluble C3 and C3b on human lymphoblastoid (RAJI) cells. Properties and biologocal significance. J Exp Med. 1974 Mar 1;139(3):696–711. doi: 10.1084/jem.139.3.696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  56. Theofilopoulos A. N., Burtonboy G., LoSpalluto J. J., Ziff M. IgM rheumatoid factor and low molecular weight IgM. An association with vasculitis. Arthritis Rheum. 1974 May-Jun;17(3):272–284. doi: 10.1002/art.1780170312. [DOI] [PubMed] [Google Scholar]
  57. Theofilopoulos A. N., Dixon F. J., Bokisch V. A. Binding of soluble immune complexes to human lymphoblastoid cells. I. Characterization of receptors for IgG Fc and complement and description of the binding mechanism. J Exp Med. 1974 Oct 1;140(4):877–894. doi: 10.1084/jem.140.4.877. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. Theofilopoulos A. N., Wilson C. B., Bokisch V. A., Dixon F. J. Binding of soluble immune complexes to human lymphoblastoid cells. II. Use of Raji cells to detect circulating immune complexes in animal and human sera. J Exp Med. 1974 Nov 1;140(5):1230–1244. doi: 10.1084/jem.140.5.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  59. Trepo C. G., Zucherman A. J., Bird R. C., Prince A. M. The role of circulating hepatitis B antigen/antibody immune complexes in the pathogenesis of vascular and hepatic manifestations in polyarteritis nodosa. J Clin Pathol. 1974 Nov;27(11):863–868. doi: 10.1136/jcp.27.11.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  60. Wands J. R., Mann E., Alpert E., Isselbacher K. J. The pathogenesis of arthritis associated with acute hepatitis-B surface antigen-positive hepatitis. Complement activation and characterization of circulating immune complexes. J Clin Invest. 1975 May;55(5):930–936. doi: 10.1172/JCI108022. [DOI] [PMC free article] [PubMed] [Google Scholar]
  61. Whitaker J. N., Engel W. K. Vascular deposits of immunoglobulin and complement in idiopathic inflammatory myopathy. N Engl J Med. 1972 Feb 17;286(7):333–338. doi: 10.1056/NEJM197202172860701. [DOI] [PubMed] [Google Scholar]
  62. Wilson C. B., Dixon F. J. Diagnosis of immunopathologic renal disease. Kidney Int. 1974 Jun;5(6):389–401. doi: 10.1038/ki.1974.57. [DOI] [PubMed] [Google Scholar]
  63. 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]
  64. Winchester R. J., Winfield J. B., Siegal F., Wernet P., Bentwich Z., Kunkel H. G. Analyses of lymphocytes from patients with rheumatoid arthritis and systemic lupus erythematosus. Occurrence of interfering cold-reactive antilymphocyte antibodies. J Clin Invest. 1974 Nov;54(5):1082–1092. doi: 10.1172/JCI107852. [DOI] [PMC free article] [PubMed] [Google Scholar]
  65. Winfield J. B., Winchester R. J., Wernet P., Fu S. M., Kunkel H. G. Nature of cold-reactive antibodies to lymphocyte surface determinants in systemic lupus erythematosus. Arthritis Rheum. 1975 Jan-Feb;18(1):1–8. doi: 10.1002/art.1780180101. [DOI] [PubMed] [Google Scholar]
  66. Winfield J. B., Winchester R. J., Wernet P., Kunkel H. G. Specific concentration of antilymphocyte antibodies in the serum cryoprecipitates of patients with systemic lupus erythematosus. Clin Exp Immunol. 1975 Mar;19(3):399–406. [PMC free article] [PubMed] [Google Scholar]
  67. Yoshiki T., Mellors R. C., Strand M., August J. T. The viral envelope glycoprotein of murine leukemia virus and the pathogenesis of immune complex glomerulonephritis of New Zealand mice. J Exp Med. 1974 Oct 1;140(4):1011–1027. doi: 10.1084/jem.140.4.1011. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES