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Infect Immun. 1972 December; 6(6): 899-904
Copyright © 1972 American Society for Microbiology. All Rights Reserved.

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

Y. Sato and H. Arai

1 First Department of Bacteriology, National Institute of Health, Kamiosaki, Shinagawa-ku, Tokyo 141, Japan

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.

Infect Immun. 1972 December; 6(6): 899-904
Copyright © 1972 American Society for Microbiology. All Rights Reserved.



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