Gastroenterologia Japonica

, Volume 15, Issue 1, pp 20–26 | Cite as

Studies on circulating immune complexes of the liver disease. 3. Conditions for Clq binding assay and low molecular weight Clq reactants

  • Terukatsu Arima
  • Jin Narumoto
  • Hiroyuki Shimomura
  • Kinichiro Suwaki
  • Kensuke Kunishi
  • Takashi Yasuhara
  • Hideo Nagashima
Original Article


Assay conditions for Clq binding activity (ClqBA) of aggregated human gamma globulin and pathologic sera were studied. The Clq component of complement was isolated by precipitation method and effectively radioiodinated using lactoperoxidase without loss of binding activity. The assay method was reproducible and the binding activity of normal sera was 5.0+-2.4% (mean +-S.D.). Amount of125I-Clq, in the range of 50–1000 ng per assay tube, did not influence the binding activity. Competitive inhibition of intrinsic Clq was very limited. Sample sera for the determination can be stored at -20‡C and freezing-thowing of the sera did not increase ClqBA. In normal sera, the existence of solubilizer of Clq-reactant-complex or inhibitor of Clqreactant binding working at 37‡C is suggested. For the low molecular weight reactants in pathologic sera reacting with Clq at 4‡C, following substances are proposed: 1. abnormal IgG, 2. hapten-IgG complexes or 3. anionic substances.

Key Words

Clq immune complexes aggregated human gamma globulin 7S-Clq reactants radioassay lactoperoxidase liver disease gel filtration 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1).
    Agnello V, Winchester RJ, Kunkel HG: Precipitin reactions of the Clq component of complement with aggregated y-globulin and immune complexes in gel diffusion. Immunology 19: 909–919, 1970PubMedGoogle Scholar
  2. 2).
    Nydegger UE, Lambert PH, Gerber H, Miescher PA: Circulating immune complexes in the serum in systemic lupus erythematosus and in carriers of hepatitis B antigen. Quantitation by binding to radiolabeled Clq. J Clin Invest 54: 297–309, 1974PubMedCrossRefGoogle Scholar
  3. 3).
    Zubler RH, Lange G, Lambert PH, Miescher PA: Detection of immune complexes in unheated sera by a modified125I-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 116:232–235, 1976PubMedGoogle Scholar
  4. 4).
    Sobel AT, Bokisch VA, Muller-Eberhard HJ: Clq deviation test for the detection of immune complexes, aggregates of IgG, and bacterial products in human serum. J Exp Med l42: 139–150, 1975CrossRefGoogle Scholar
  5. 5).
    Svehag SE: A solid-phase radioimmunoassay for Clqbinding immune complexes. 1. δIgG as indicator molecule. Scand J Immunol 4: 687–697, 1975PubMedCrossRefGoogle Scholar
  6. 6).
    Farrell C, SØgard H, Svehag SE: Detection of IgG aggregates or immune complexes using solid-phase Clq and protein A-rich staphylococcus aureus as an indicator system. Scand J Immunol 4: 673–680, 1975PubMedCrossRefGoogle Scholar
  7. 7).
    Lurhuma AZ, Cambiaso CL, Masson PL, Heremans JF: Detection of circulating antigen-antibody complexes by their inhibitory effect on the agglutination of IgG-coated particles by rheumatoid factor or Clq. Clin Exp Immunol 25: 212–226, 1976PubMedGoogle Scholar
  8. 8).
    Hay FC, Nineham LJ, Roitt IM: Routine assay for the detection of immune complexes of known immunoglobulin class using solid phase Clq. Clin Exp Immunol 24:396–400, 1976PubMedGoogle Scholar
  9. 9).
    Ahlstedt S, Hanson LA, Wadsworth C: A Clq immunosorbent assay compared with thin-layer gel filtration for measuring IgG aggregates. Scand J Immunol 5: 293–298, 1976PubMedCrossRefGoogle Scholar
  10. 10).
    Fletcher DS, Lin T: Quantitation of immune complexes by competitive inhibition of binding of Clq to insoluble IgG aggregates. J Immunol Methods 15: 39–45, 1977PubMedCrossRefGoogle Scholar
  11. 11).
    Gabriel A, Agnello V: Detection of immune complexes. The use of radioimmunoassays with Clq and monoclonal rheumatoid factor. J Clin Invest 59: 990–1001, 1977PubMedGoogle Scholar
  12. 12).
    Agnello V, Koffler D, Eisenberg JW, Winchester RJ, Kunkel HG: Clq 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 134: 228s-240s, 1971PubMedGoogle Scholar
  13. 13).
    Yonemasu K, Stroud RM: Clq: rapid purification method for preparation of monospecific antisera and for biochemical studies. J Immunol 106: 304–313, 1971PubMedGoogle Scholar
  14. 14).
    Arima T, Miyamoto-Sudo C, Hirohata M, Tanigawa T, Tsuboi S, Tsunajima T, Kuwauchi S, Imai M: Further studies on an eleventh case of heavy (Hγl) chain disease-clinical studies. Acta Med Okayama 29:199–208, 1975PubMedGoogle Scholar
  15. 15).
    Morrison M, Bayse GS, Webster RG: Use of lactoperoxidase catalyzed iodination in immunochemical studies. Immunochemistry 8: 289–297, 1971PubMedCrossRefGoogle Scholar
  16. 16).
    Heusser C, Boesman M, Nordin JM, Isliker H: Effect of chemical and enzymatic iodination on in vitro human Clq activities. J Immunol 110: 820–828, 1973PubMedGoogle Scholar
  17. 17).
    Arima T, Miyamoto-Sudo C, Hirohata M, Tanigawa T, Tsuboi S, Tsunajima T, Kuwauchi S, Imai M: Further studies on an eleventh case of heavy (Hγl) chain disease-physico-chemical studies. Acta Med Okayama 29: 209–223, 1975PubMedGoogle Scholar
  18. 18).
    Arima T, Miyamoto-Sudo C, Hirohata M, Tsunajima T, Tsuboi S, Tsunajima T, Kuwauchi S, Imai M: Further studies on an eleventh case of heavy (Hγl) chain disease-biosynthetic studies. Acta Med Okayama 29: 225–231, 1975PubMedGoogle Scholar
  19. 19).
    Miller GW, Nussenzweig V: A new complement function: solubilization of antigen-antibody aggregates. Proc Nat Acad Sci USA 72: 418–422, 1975PubMedCrossRefGoogle Scholar
  20. 20).
    Czop J, Nussenzweig V: Studies on the mechanism of solubilization of immune precipitates by serum. J Exp Med 143: 615–630, 1976PubMedCrossRefGoogle Scholar
  21. 21).
    Conradie JD, Valanakis JE, Stroud RM: Evidence for a serum inhibitor of Clq. Immunochemistry 12: 967–971, 1975PubMedCrossRefGoogle Scholar
  22. 22).
    Calcott MA, Muller-Eberhard HJ: Clq protein of human complement. Biochemistry 11: 3443–3450, 1972PubMedCrossRefGoogle Scholar
  23. 23).
    Marder RJ, Burch FX, Schmid FR, Zeiss CR, Gewurz H: Low molecular weight Clq-precipitins in hypocomplementemic vasculitis-urticaria syndrome: partial purification and characterization as immunoglobulin. J Immunol 121: 613–618, 1978PubMedGoogle Scholar
  24. 24).
    Muller-Eberhard HJ, Kunkel HG: Isolation of a thermolabile serum protein which precipitates γglobulin aggregates and participates in immune hemolysis. Proc Soc Exp Biol 106: 291–295, 1961Google Scholar
  25. 25).
    Cohen S: The requirement for the association of two adjacent rabbit γG-antibody molecules in the fixation of complement by immune complexes. J Immunol 100:407–413, 1968PubMedGoogle Scholar
  26. 26).
    Augener W, Grey HM, Cooper NR, Muller-Eberhard HJ: The reaction of monomeric and aggregated immunoglobulins with Cl. Immunochemistry 8: 1011–1020, 1971PubMedCrossRefGoogle Scholar
  27. 27).
    Sledge CR, Bing DH: Binding properties of the human complement protein Clq. J Biol Chem 248: 2818–2823, 1973PubMedGoogle Scholar
  28. 28).
    Lin T, Fletcher DS: Interaction of human Clq with insoluble immunoglobulin aggregates. Immunochemistry 15: 107–117, 1978PubMedCrossRefGoogle Scholar
  29. 29).
    Goers JW, Schumaker VN, Glovsky MM, Rebek J, Muller-Eberhard HJ: Complement activation by a univalent hapten-antibody complex. J Biol Chem 250: 4918–4925, 1975PubMedGoogle Scholar

Copyright information

© The Japanese Society of Gastroenterology 1980

Authors and Affiliations

  • Terukatsu Arima
    • 1
  • Jin Narumoto
    • 1
  • Hiroyuki Shimomura
    • 1
  • Kinichiro Suwaki
    • 1
  • Kensuke Kunishi
    • 1
  • Takashi Yasuhara
    • 1
  • Hideo Nagashima
    • 1
  1. 1.The First Department of Internal MedicineOkayama University Medical SchoolOkayamaJapan

Personalised recommendations