Abstract
Nephritic factor (C3NeF) has been isolated from plasma of patients with hypocomplementemic chronic glomerulonephritis (HCG) by ion exchange and molecular sieve chromatography. This material was further treated with solidified anti-Ig antiserum. The purified material failed to react with antiserum to human IgG, IgG3, Fab, Fc, and kappa and lambda chains, but retained full C3NeF activity. The nonidentity of C3NeF with IgG was further demonstrated by Ouchterlony analysis using anti-IgG and anti-C3NeF. Isolated C3NeF was found to be a protein with a sedimentation coefficient of 7S and a mol wt of 150,000 daltons, which on microzone electrophoresis and gel electrophoresis at pH 8.6 behaved as a γ-globulin. C3NeF is not a C1q precipitin and does not activate the classical complement pathway. Unlike cobra venom factor, it failed to enter into a complex with C3 proactivator (C3PA) when incubated with normal human serum (NHS) and then subjected to sucrose density gradient ultracentrifugation. The action of isolated C3NeF on C3 requires C3PA, C3PA convertase (C3PAse), and properdin (P). Similarly, C3PA conversion by C3NeF requires P, C3PAse, and C3. Total hemolytic activity was lost by incubation of 64 µg of C3NeF/1 ml NHS at 37°C for 30 min. Both C3a and C5a anaphylatoxin could be generated by C3NeF in serum previously depleted of anaphylatoxin inactivator. Anti-C3NeF was found to detect an antigen in all NHS tested. Treatment of NHS with solidified anti-C3NeF caused impairment of the alternate complement pathway. It failed to sustain lysis of glutathione-treated human erythrocytes initiated by inulin. It is conceivable that the normal serum constituent which is removed by anti-C3NeF constitutes the inactive precursor of C3NeF, and a heretofore unrecognized component of the alternate pathway.
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- Avrameas S., Ternynck T. Biologically active water-insoluble protein polymers. I. Their use for isolation of antigens and antibodies. J Biol Chem. 1967 Apr 10;242(7):1651–1659. [PubMed] [Google Scholar]
- Cochrane C. G., Müller-Eberhard H. J. The derivation of two distinct anaphylatoxin activities from the third and fifth components of human complement. J Exp Med. 1968 Feb 1;127(2):371–386. doi: 10.1084/jem.127.2.371. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Day N. K., Geiger H., McLean R., Resnick J., Michael A., Good R. A. The association of respiratory infection, recurrent hematuria, and focal glomerulonephritis with activation of the complement system in the cold. J Clin Invest. 1973 Jul;52(7):1698–1706. doi: 10.1172/JCI107351. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Edgington T. S. Dissociation of antibody from erythrocyte surfaces by chaotropic ions. J Immunol. 1971 Mar;106(3):673–680. [PubMed] [Google Scholar]
- GOTOFF S. P., FELLERS F. X., VAWTER G. F., JANEWAY C. A., ROSEN F. S. THE BETA-1C GLOBULIN IN CHILDHOOD NEPHROTIC SYNDROME: LABORATORY DIAGNOSIS OF PROGRESSIVE GLOMERULONEPHRITIS. N Engl J Med. 1965 Sep 2;273:524–529. doi: 10.1056/NEJM196509022731004. [DOI] [PubMed] [Google Scholar]
- Hunsicker L. G., Ruddy S., Carpenter C. B., Schur P. H., Merrill J. P., Müller-Eberhard H. J., Austen K. F. Metabolism of third complement component (C3) in nephritis. Involvement of the classic and alternate (properdin) pathways for complement activation. N Engl J Med. 1972 Oct 26;287(17):835–840. doi: 10.1056/NEJM197210262871701. [DOI] [PubMed] [Google Scholar]
- 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]
- McLean R. H., Michael A. F. Properdin anc C3 proactivator: alternate pathway components in human glomerulonephritis. J Clin Invest. 1973 Mar;52(3):634–644. doi: 10.1172/JCI107225. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ruley E. J., Forristal J., Davis N. C., Andres C., West C. D. Hypocomplementemia of membranoproliferative nephritis. Dependence of the nephritic factor reaction on properdin factor B. J Clin Invest. 1973 Apr;52(4):896–904. doi: 10.1172/JCI107254. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thompson R. A. C3 inactivating factor in the serum of a patient with chronic hypocomplementaemic proliferative glomerulo-nephritis. Immunology. 1972 Jan;22(1):147–158. [PMC free article] [PubMed] [Google Scholar]
- Vallota E. H., Forristal J., Davis N. C., West C. D. The C3 nephritic factor and membranoproliferative nephritis: correlation of serum levels of the nephritic factor with C3 levels, with therapy, and with progression of the disease. J Pediatr. 1972 Jun;80(6):947–959. doi: 10.1016/s0022-3476(72)80006-4. [DOI] [PubMed] [Google Scholar]
- Vallota E. H., Forristal J., Spitzer R. E., Davis N. C., West C. D. Characteristics of a non-complement-dependent C3-reactive complex formed form factors in nephritic and normal serum. J Exp Med. 1970 Jun 1;131(6):1306–1324. doi: 10.1084/jem.131.6.1306. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vallota E. H., Forristal J., Spitzer R. E., Davis N. C., West C. D. Continuing C3 breakdown after bilateral nephrectomy in patients with membrano-proliferative glomerulonephritis. J Clin Invest. 1971 Mar;50(3):552–558. doi: 10.1172/JCI106524. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vallota E. H., Müller-Eberhard H. J. Formation of C3a and C5a anaphylatoxins in whole human serum after inhibition of the anaphylatoxin inactivator. J Exp Med. 1973 May 1;137(5):1109–1123. doi: 10.1084/jem.137.5.1109. [DOI] [PMC free article] [PubMed] [Google Scholar]
- West C. D., Winter S., Forristal J., McConville J. M., Davis N. C. Evidence for in vivo breakdown of beta-10-globulin in hypocomplementemic glomerulonephritis. J Clin Invest. 1967 Apr;46(4):539–548. doi: 10.1172/JCI105555. [DOI] [PMC free article] [PubMed] [Google Scholar]