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. 1972 Dec;6(6):899–904. doi: 10.1128/iai.6.6.899-904.1972

Leucocytosis-Promoting Factor of Bordetella pertussis I. Purification and Characterization

Y Sato 1, H Arai 1
PMCID: PMC419309  PMID: 4344632

Abstract

The leucocytosis-promoting factor was purified from the supernatant fluid of spent cultures of Bordetella pertussis on solid medium. After precipitation at 67% saturation of ammonium sulfate, the leucocytosis-promoting factor was extracted with a 1.0 m NaCl solution. Purification was accomplished by starch block electrophoresis and sucrose density gradient centrifugation. The purified preparation contained a high leucocytosis-promoting activity, and as small an amount as 0.04 μg of protein induced leucocytosis in mice. About 520-fold purification was attained, with a re-recovery of about 25% on an activity basis. The leucocytosis-promoting factor was composed solely of filamentous molecules of about 2 by 40 nm in size, with a sedimentation coefficient of approximately 5.5S and a molecular weight of 108,000. It was insoluble in water but partially soluble in 1.0 m NaCl solution, and consisted mainly of protein, with some carbohydrate, lipid, and phosphorus.

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Selected References

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

  1. Allen R. J. The estimation of phosphorus. Biochem J. 1940 Jun;34(6):858–865. doi: 10.1042/bj0340858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. BURTON K. A study of the conditions and mechanism of the diphenylamine reaction for the colorimetric estimation of deoxyribonucleic acid. Biochem J. 1956 Feb;62(2):315–323. doi: 10.1042/bj0620315. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Clausen C., Munoz J., Bergman R. K. Lymphocytosis and histamine sensitization of mice by fractions from Bordetella pertussis. J Bacteriol. 1968 Nov;96(5):1484–1487. doi: 10.1128/jb.96.5.1484-1487.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DISCHE Z., BORENFREUND E. A spectrophotometric method for the microdetermination of hexosamines. J Biol Chem. 1950 Jun;184(2):517–522. [PubMed] [Google Scholar]
  5. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  6. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  7. MARTIN R. G., AMES B. N. A method for determining the sedimentation behavior of enzymes: application to protein mixtures. J Biol Chem. 1961 May;236:1372–1379. [PubMed] [Google Scholar]
  8. MORSE S. I. STUDIES ON THE LYMPHOCYTOSIS INDUCED IN MICE BY BORDETELLA PERTUSSIS. J Exp Med. 1965 Jan 1;121:49–68. doi: 10.1084/jem.121.1.49. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Morse J. H., Morse S. I. Studies on the ultrastructure of Bordetella pertussis. I. Morphology, origin, and biological activity of structures present in the extracellular fluid of liquid cultures of Bordetella pertussis. J Exp Med. 1970 Jun 1;131(6):1342–1357. doi: 10.1084/jem.131.6.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Morse S. I., Barron B. A. Studies on the leukocytosis and lymphocytosis induced by Bordetella pertussis. 3. The distribution of transfused lymphocytes in pertussis-treated and normal mice. J Exp Med. 1970 Oct 1;132(4):663–672. doi: 10.1084/jem.132.4.663. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Morse S. I., Bray K. K. The occurrence and properties of leukocytosis and lymphocytosis-stimulating material in the supernatant fluids of Bordetella pertussis cultures. J Exp Med. 1969 Mar 1;129(3):523–550. doi: 10.1084/jem.129.3.523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Morse S. I., Riester S. K. Studies on the leukocytosis and lymphocytosis induced by Bordetella pertussis. I. Radioautographic analysis of the circulating cells in mice undergoing pertussis-induced hyperleukocytosis. J Exp Med. 1967 Mar 1;125(3):401–408. doi: 10.1084/jem.125.3.401. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Morse S. I., Riester S. K. Studies on the leukocytosis and lymphocytosis induced by Bordetella pertussis. II. The effect of pertussis vaccine on the thoracic duct lymph and lymphocytes of mice. J Exp Med. 1967 Apr 1;125(4):619–628. doi: 10.1084/jem.125.4.619. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Munoz J., Bergman R. K. Histamine-sensitizing factors from microbial agents, with special reference to Bordetella pertussis. Bacteriol Rev. 1968 Jun;32(2):103–126. doi: 10.1128/br.32.2.103-126.1968. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Okuyama S., Aronson R. B., Chanana A. D., Cronkite E. P., Rai K. R., Schiffer L. M. Attempts at isolation of lymphocytosis-producing factor from supernatant fluids of Bordetella pertussis cultures. Proc Soc Exp Biol Med. 1970 Feb;133(2):723–727. [PubMed] [Google Scholar]
  16. Sato Y., Arai H. Partial purification and some properties of the leukocytosis-promoting material from Bordetella pertussis. Jpn J Med Sci Biol. 1970 Aug;23(4):273–276. [PubMed] [Google Scholar]
  17. TOENNIES G., KOLB J. J. CARBOHYDRATE ANALYSIS OF BACTERIAL SUBSTANCES BY A NEW ANTHRONE PROCEDURE. Anal Biochem. 1964 May;8:54–69. doi: 10.1016/0003-2697(64)90168-x. [DOI] [PubMed] [Google Scholar]
  18. Uiwasa S., Ishida S., Asakawa S., Kurokawa M. Lymphocytosis-promoting factor produced by Bordetella pertussis. Partial purification and characterization. Jpn J Med Sci Biol. 1968 Oct;21(5):363–368. doi: 10.7883/yoken1952.21.363. [DOI] [PubMed] [Google Scholar]
  19. WEBB J. M. A sensitive method for the determination of ribonucleic acid in tissues and microorganisms. J Biol Chem. 1956 Aug;221(2):635–649. [PubMed] [Google Scholar]

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