Abstract
Only antibodies of the IgM class support the lytic effect of complement on Giardia lamblia (GL). We sensitized GL trophozoites (SGL) at 4 degrees C with serum containing anti-GL antibodies or IgM purified from this serum, and either normal human serum (NHS), complement 2-deficient human serum (C2d-HS), or C4-deficient guinea pig serum was used as source of complement. SGL were killed by NHS (86%) and by the deficient sera (50 and 40%, respectively), suggesting activation of the alternative pathway. However, the reaction was inhibited by Mg-EGTA. These observations led to studies of the role of C1. The lytic effect of NHS and C2d-HS on SGL was abolished by immunochemically depleting C1 from these sera, and reconstituted by adding purified C1q plus C1r and C1s. Factor B-depleted C2d-HS also lost its capacity to mediate killing, but reconstitution with factor B led to a dose-dependent increase in the killing of SGL. We next investigated the participation of the membrane attack complex in this system. SGL carrying C5b to C7 were lysed when incubated with C8 alone (56%); the addition of C9 further increased killing (98%), while C9 in the absence of C8 had no effect. We concluded that although activation of the classical pathway produces lysis of SGL, lysis may also proceed through a unique pathway of complement activation that requires C1 and factor B, but is independent of C4 and C2. Lysis of SGL can be accomplished by C5b to C8 in the absence of C9.
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- BORSOS T., RAPP H. J. CHROMATOGRAPHIC SEPARATION OF THE FIRST COMPONENT OF COMPLEMENT AND ITS ASSAY ON A MOLECULAR BASIS. J Immunol. 1963 Dec;91:851–858. [PubMed] [Google Scholar]
- Boyle M. D., Ohanian S. H., Borsos T. Lysis of tumor cells by antibody and complement. VII. Complement-dependent 86Rb release--a nonlethal event? J Immunol. 1976 Oct;117(4):1346–1350. [PubMed] [Google Scholar]
- Carney D. F., Koski C. L., Shin M. L. Elimination of terminal complement intermediates from the plasma membrane of nucleated cells: the rate of disappearance differs for cells carrying C5b-7 or C5b-8 or a mixture of C5b-8 with a limited number of C5b-9. J Immunol. 1985 Mar;134(3):1804–1809. [PubMed] [Google Scholar]
- Diamond L. S., Harlow D. R., Cunnick C. C. A new medium for the axenic cultivation of Entamoeba histolytica and other Entamoeba. Trans R Soc Trop Med Hyg. 1978;72(4):431–432. doi: 10.1016/0035-9203(78)90144-x. [DOI] [PubMed] [Google Scholar]
- Fraker P. J., Speck J. C., Jr Protein and cell membrane iodinations with a sparingly soluble chloroamide, 1,3,4,6-tetrachloro-3a,6a-diphrenylglycoluril. Biochem Biophys Res Commun. 1978 Feb 28;80(4):849–857. doi: 10.1016/0006-291x(78)91322-0. [DOI] [PubMed] [Google Scholar]
- Harriman G. R., Esser A. F., Podack E. R., Wunderlich A. C., Braude A. I., Lint T. F., Curd J. G. The role of C9 in complement-mediated killing of Neisseria. J Immunol. 1981 Dec;127(6):2386–2390. [PubMed] [Google Scholar]
- Hermans P. E., Diaz-Buxo J. A., Stobo J. D. Idiopathic late-onset immunoglobulin deficiency. Clinical observations in 50 patients. Am J Med. 1976 Aug;61(2):221–237. doi: 10.1016/0002-9343(76)90173-x. [DOI] [PubMed] [Google Scholar]
- Hill D. R., Burge J. J., Pearson R. D. Susceptibility of Giardia lamblia trophozoites to the lethal effect of human serum. J Immunol. 1984 Apr;132(4):2046–2052. [PubMed] [Google Scholar]
- Keister D. B. Axenic culture of Giardia lamblia in TYI-S-33 medium supplemented with bile. Trans R Soc Trop Med Hyg. 1983;77(4):487–488. doi: 10.1016/0035-9203(83)90120-7. [DOI] [PubMed] [Google Scholar]
- Kitamura F., Shimada K., Suzuki T., Nishioka K. A simplified method for the preparation of EAC14 intermediate cells using human serum treated with methylprednisolone. J Immunol Methods. 1985 Dec 27;85(2):363–370. doi: 10.1016/0022-1759(85)90145-0. [DOI] [PubMed] [Google Scholar]
- LEPOW I. H., NAFF G. B., TODD E. W., PENSKY J., HINZ C. F. Chromatographic resolution of the first component of human complement into three activities. J Exp Med. 1963 Jun 1;117:983–1008. doi: 10.1084/jem.117.6.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lesavre P. H., Hugli T. E., Esser A. F., Müller-Eberhard H. J. The alternative pathway C3/C5 convertase: chemical basis of factor B activation. J Immunol. 1979 Aug;123(2):529–534. [PubMed] [Google Scholar]
- Lint T. F., Zeitz H. J., Gewurz H. Inherited deficiency of the ninth component of complement in man. J Immunol. 1980 Nov;125(5):2252–2257. [PubMed] [Google Scholar]
- May J. E., Frank M. M. A new complement-mediated cytolytic mechanism--the C1-bypass activation pathway. Proc Natl Acad Sci U S A. 1973 Mar;70(3):649–652. doi: 10.1073/pnas.70.3.649. [DOI] [PMC free article] [PubMed] [Google Scholar]
- May J. E., Frank M. M. Hemolysis of sheep erythrocytes in guinea pig serum deficient in the fourth component of complement. II. Evidence for involvement of C1 and components of the alternate complement pathway. J Immunol. 1973 Dec;111(6):1668–1676. [PubMed] [Google Scholar]
- Morgan B. P., Imagawa D. K., Dankert J. R., Ramm L. E. Complement lysis of U937, a nucleated mammalian cell line in the absence of C9: effect of C9 on C5b-8 mediated cell lysis. J Immunol. 1986 May 1;136(9):3402–3406. [PubMed] [Google Scholar]
- Ohanian S. H., Schlager S. I. Humoral immune killing of nucleated cells: mechanisms of complement-mediated attack and target cell defense. Crit Rev Immunol. 1981 Jan;1(3):165–209. [PubMed] [Google Scholar]
- Platts-Mills T. A., Ishizaka K. Activation of the alternate pathway of human complements by rabbit cells. J Immunol. 1974 Jul;113(1):348–358. [PubMed] [Google Scholar]
- Ramm L. E., Whitlow M. B., Koski C. L., Shin M. L., Mayer M. M. Elimination of complement channels from the plasma membranes of U937, a nucleated mammalian cell line: temperature dependence of the elimination rate. J Immunol. 1983 Sep;131(3):1411–1415. [PubMed] [Google Scholar]
- Ramm L. E., Whitlow M. B., Mayer M. M. Size of the transmembrane channels produced by complement proteins C5b-8. J Immunol. 1982 Sep;129(3):1143–1146. [PubMed] [Google Scholar]
- Ridley M. J., Ridley D. S. Serum antibodies and jejunal histology in giardiasis associated with malabsorption. J Clin Pathol. 1976 Jan;29(1):30–34. doi: 10.1136/jcp.29.1.30. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schreiber R. D., Pangburn M. K., Medicus R. G., Müller-Eberhard H. J. Raji cell injury and subsequent lysis by the purified cytolytic alternative pathway of human complement. Clin Immunol Immunopathol. 1980 Mar;15(3):384–396. doi: 10.1016/0090-1229(80)90050-1. [DOI] [PubMed] [Google Scholar]
- Smith P. D., Elson C. O., Keister D. B., Nash T. E. Human host response to Giardia lamblia. I. Spontaneous killing by mononuclear leukocytes in vitro. J Immunol. 1982 Mar;128(3):1372–1376. [PubMed] [Google Scholar]
- Smith P. D., Gillin F. D., Brown W. R., Nash T. E. IgG antibody to Giardia lamblia detected by enzyme-linked immunosorbent assay. Gastroenterology. 1981 Jun;80(6):1476–1480. [PubMed] [Google Scholar]
- Stolfi R. L. Immune lytic transformation: a state of irreversible damage generated as a result of the reaction of the eighth component in the guinea pig complement system. J Immunol. 1968 Jan;100(1):46–54. [PubMed] [Google Scholar]
- Tamura N., Shimada A., Chang S. Further evidence for immune cytolysis by antibody and the first eight components of complement in the absence of C9. Immunology. 1972 Jan;22(1):131–140. [PMC free article] [PubMed] [Google Scholar]
- Thompson A., Rowland R., Hecker R., Gibson G. E., Reid D. P. Immunoglobulin-bearing cells in giardiasis. J Clin Pathol. 1977 Mar;30(3):292–294. doi: 10.1136/jcp.30.3.292. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Visvesvara G. S., Smith P. D., Healy G. R., Brown W. R. An immunofluorescence test to detect serum antibodies to Giardia lamblia. Ann Intern Med. 1980 Dec;93(6):802–805. doi: 10.7326/0003-4819-93-6-802. [DOI] [PubMed] [Google Scholar]