Skip to main content
Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1983 Aug;18(2):248–253. doi: 10.1128/jcm.18.2.248-253.1983

Determination of serum bactericidal activity with the aid of luminous bacteria.

M Barak, S Ulitzur, D Merzbach
PMCID: PMC270786  PMID: 6619281

Abstract

Nonmarine luminous bacteria belonging to the genus Vibrio cholerae were extremely sensitive to the bactericidal activity of human serum. Luminous bacteria incubated in a medium containing serum showed a decrease in their in vivo luminescence that was directly proportional to the decrease in the viable count and was a function of the serum concentration. Both immunoglobulins and the complement system were required to exert the serum bactericidal activity. Serum lacking immunoglobulins or certain complement components, especially C3, did not affect the luminescence. The bactericidal effect of the serum on luminous bacteria was diminished by the presence of lipopolysaccharide or by pretreatment of the serum with different species of killed bacteria. As found in other systems, the bacteriolytic activity of serum was only augmented by lysozyme, but was not lysozyme dependent; although the luminous bacteria were converted into spheroplasts in serum containing 0.5 M sucrose, their in vivo luminescence was almost not affected. This system could easily distinguish between the C classical pathway and the properdin pathway. Ethylene glycol-bis (beta-aminoethyl ether)-N,N'-tetraacetic acid, which inhibits only the classical complement pathway, did not inhibit the decrease in luminescence as did EDTA. Thus, it was possible to distinguish between deficiencies in complement components participating in both pathways and complement components that were involved only in the classical pathway. This system could also be used as a substitute to the hemolytic system in complement fixation tests.

Full text

PDF
248

Selected References

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

  1. Allen R. J., Scott G. K. Comparison of the effects of different lipopolysaccharides on the serum bactericidal reactions of two strains of Escherichia coli. Infect Immun. 1981 Feb;31(2):831–832. doi: 10.1128/iai.31.2.831-832.1981. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allen R. J., Scott G. K. Human serum complement requirements for bacterial killing and protoplast lysis of Escherichia coli ML308 225. J Gen Microbiol. 1981 Mar;123(1):179–181. doi: 10.1099/00221287-123-1-179. [DOI] [PubMed] [Google Scholar]
  3. Allen R. J., Scott G. K. The effect of purified lipopolysaccharide on the bactericidal reaction of human serum complement. J Gen Microbiol. 1980 Mar;117(1):65–72. doi: 10.1099/00221287-117-1-65. [DOI] [PubMed] [Google Scholar]
  4. Davis S. D., Gemsa D., Wedgwood R. J. Kinetics of the transformation of Gram-negative rods to spheroplasts and ghosts by serum. J Immunol. 1966 Apr;96(4):570–577. [PubMed] [Google Scholar]
  5. Day N. K., Geiger H., McLean R., Michael A., Good R. A. C2 deficiency. Development of lupus erythematosus. J Clin Invest. 1973 Jul;52(7):1601–1607. doi: 10.1172/JCI107337. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Fine D. P., Marney S. R., Jr, Colley D. G., Sergent J. S., Des Prez R. M. C3 shunt activation in human serum chelated with EGTA. J Immunol. 1972 Oct;109(4):807–809. [PubMed] [Google Scholar]
  7. Glynn A. A., Milne C. M. A kinetic study of the bacteriolytic and bactericidal action of human serum. Immunology. 1967 Jun;12(6):639–653. [PMC free article] [PubMed] [Google Scholar]
  8. Goldman J. N., Ruddy S., Austen K. F., Feingold D. S. The serum bactericidal reaction. 3. Antibody and complement requirements for killing a rough Escherichia coli. J Immunol. 1969 Jun;102(6):1379–1387. [PubMed] [Google Scholar]
  9. Jørgensen K. A., Stoffersen E. The complement system in uremia. Scand J Urol Nephrol. 1980;14(3):279–282. doi: 10.3109/00365598009179576. [DOI] [PubMed] [Google Scholar]
  10. Lim D., Gewurz A., Lint T. F., Ghaze M., Sepheri B., Gewurz H. Absence of the sixth component of complement in a patient with repeated episodes of meningococcal meningitis. J Pediatr. 1976 Jul;89(1):42–47. doi: 10.1016/s0022-3476(76)80924-9. [DOI] [PubMed] [Google Scholar]
  11. MUSCHEL L. H., JACKSON J. E. ACTIVITY OF THE ANTIBODY-COMPLEMENT SYSTEM AND LYSOZYME AGAINST ROUGH GRAM NEGATIVE ORGANISMS. Proc Soc Exp Biol Med. 1963 Aug-Sep;113:881–884. doi: 10.3181/00379727-113-28519. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Martinez R. J., Carroll S. F. Sequential metabolic expressions of the lethal process in human serum-treated Escherichia coli: role of lysozyme. Infect Immun. 1980 Jun;28(3):735–745. doi: 10.1128/iai.28.3.735-745.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Melching L., Vas S. I. Effects of serum components on gram-negative bacteria during bactericidal reactions. Infect Immun. 1971 Jan;3(1):107–115. doi: 10.1128/iai.3.1.107-115.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mitchell G. W., Hastings J. W. A stable, inexpensive, solid-state photomultiplier photometer. Anal Biochem. 1971 Jan;39(1):243–250. doi: 10.1016/0003-2697(71)90481-7. [DOI] [PubMed] [Google Scholar]
  16. Müller-Eberhard H. J. Complement. Annu Rev Biochem. 1975;44:697–724. doi: 10.1146/annurev.bi.44.070175.003405. [DOI] [PubMed] [Google Scholar]
  17. Ross I. N., Thompson R. A., Montgomery R. D., Asquith P. Significance of serum complement levels in patients with gastrointestinal disease. J Clin Pathol. 1979 Aug;32(8):798–801. doi: 10.1136/jcp.32.8.798. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Wilson B. M., Glynn A. A. Release of 14C label and complement killing of Escherichia coli. Immunology. 1975 Mar;28(3):391–400. [PMC free article] [PubMed] [Google Scholar]
  20. Wilson L. A., Spitznagel J. K. Molecular and structural damage to Escherichia coli produced by antibody, complement, and lysozyme systems. J Bacteriol. 1968 Oct;96(4):1339–1348. doi: 10.1128/jb.96.4.1339-1348.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Winkelstein J. A., Kurlandsky L. E., Swift A. J. Defective activation of the third component of complement in the sera of newborn infants. Pediatr Res. 1979 Oct;13(10):1093–1096. doi: 10.1203/00006450-197910000-00001. [DOI] [PubMed] [Google Scholar]
  22. Wyatt R. J., McAdams A. J., Forristal J., Snyder J., West C. D. Glomerular deposition of complement-control proteins in acute and chronic glomerulonephritis. Kidney Int. 1979 Oct;16(4):505–512. doi: 10.1038/ki.1979.156. [DOI] [PubMed] [Google Scholar]
  23. Zeitz H. J., Miller G. W., Lint T. F., Ali M. A., Gewurz H. Deficiency of C7 with systemic lupus erythematosus: solubilization of immune complexes in complement-deficient sera. Arthritis Rheum. 1981 Jan;24(1):87–93. doi: 10.1002/art.1780240114. [DOI] [PubMed] [Google Scholar]
  24. van Dijk H., Rademaker P. M., Willers J. M. Estimation of classical pathway of mouse complement activity by use of sensitized rabbit erythrocytes. J Immunol Methods. 1980;39(3):257–268. doi: 10.1016/0022-1759(80)90060-5. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES