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. 1963 Oct 31;118(5):795–815. doi: 10.1084/jem.118.5.795

COMPONENTS OF GUINEA PIG COMPLEMENT

II. SEPARATION OF SERUM FRACTIONS ESSENTIAL FOR IMMUNE HEMOLYSIS

William D Linscott 1, Kusuya Nishioka 1
PMCID: PMC2137680  PMID: 14087622

Abstract

The elution characteristics from DEAE cellulose are presented for four components of guinea pig serum, which are capable of interacting sequentially with sheep erythrocytes sensitized with antibody and the first, fourth, and second components of complement (EAC'1,4,2) to cause immune hemolysis, and information is given regarding some of the properties of these components, termed C'3c, C'3b, C'3a, and C'3d. All can react in the presence of ethylene-diaminetetraacetate, and are non-dialyzable. C'3c is quite stable at 56°C, but is rapidly inactivated at low pH or by contact with hydrazine or ammonium hydroxide. C'3b is moderately heat-stable, quite susceptible to low pH, and less readily destroyed by hydrazine. C'3a is very heat-labile, but relatively stable at low pH, while C'3d is heat-labile, sensitive to low pH, and insensitive to bydrazine. EAC'1,4,2 reacts with C'3c to form EAC'1,4,2,3c, which reacts then with C'3b to give the intermediate, EAC'1,4,2,3cb. The following reaction with C'3a yields EAC'1,4,2,3cba, which reacts finally with C'3d to give EAC'1,4, 2,3cbad (E*). The first and last reactions proceed moderately well at 0°C, but more rapidly at 30–37°C. The reaction with C'3b is almost completely inhibited at 0°C, while that involving C'3a proceeds almost as rapidly at 0°C as at higher temperatures. EAC'1,4,2,3cba cells have an increased fragility as compared with the other intermediate forms. Depletion studies with purified fractions and appropriate intermediate complexes showed a high degree of depletion of C'3c, somewhat less of C'3b, and little or no depletion of C'3a from the fluid phase. Examination of a beta1C globulin prepared from fresh human serum revealed high C'3c and C'3b activity, and very little C'3a or C'3d.

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Selected References

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

  1. MULLER-EBERHARD H. J. Isolation and description of proteins related to the human complement system. Acta Soc Med Ups. 1961;66:152–170. [PubMed] [Google Scholar]
  2. MUSCHEL L. H., CAREY W. F., BARON L. S. Formation of bacterial protoplasts by serum components. J Immunol. 1959 Jan;82(1):38–42. [PubMed] [Google Scholar]
  3. Müller-Eberhard H. J., Nilsson U. RELATION OF A beta(1)-GLYCOPROTEIN OF HUMAN SERUM TO THE COMPLEMENT SYSTEM. J Exp Med. 1960 Jan 31;111(2):217–234. doi: 10.1084/jem.111.2.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. NELSON R. A., Jr An alternative mechanism for the properdin system. J Exp Med. 1958 Oct 1;108(4):515–535. doi: 10.1084/jem.108.4.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. NISHIOKA K., LINSCOTT W. D. COMPONENTS OF GUINEA PIG COMPLEMENT. I. SEPARATION OF A SERUM FRACTION ESSENTIAL FOR IMMUNE HEMOLYSIS AND IMMUNE ADHERENCE. J Exp Med. 1963 Nov 1;118:767–793. doi: 10.1084/jem.118.5.767. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. RAPP H. J., SIMS M. R., BORSOS T. Separation of components of guinea pig complement by chromatography. Proc Soc Exp Biol Med. 1959 Apr;100(4):730–733. doi: 10.3181/00379727-100-24758. [DOI] [PubMed] [Google Scholar]
  7. TAYLOR A. B., LEON M. A. Third component of human complement: resolution into two factors and demonstration of a new reaction intermediate. Proc Soc Exp Biol Med. 1959 Jul;101(3):587–588. doi: 10.3181/00379727-101-25026. [DOI] [PubMed] [Google Scholar]
  8. WARDLAW A. C. The complement-dependent bacteriolytic activity of normal human serum. I. The effect of pH and ionic strength and the role of lysozyme. J Exp Med. 1962 Jun 1;115:1231–1249. doi: 10.1084/jem.115.6.1231. [DOI] [PMC free article] [PubMed] [Google Scholar]

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