The Journal of Experimental Medicine
for flow cytometry > invitrogen
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Published 7 September 2004. doi:10.1084/jem.20040725
Rockefeller University Press, 0022-1007 $8.00
JEM, Volume 200, Number 5, 613-622
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Determination of the Critical Concentration of Neutrophils Required to Block Bacterial Growth in Tissues

Yongmei Li1, Arthur Karlin1, John D. Loike1, and Samuel C. Silverstein1,2

1 Department of Physiology and Cellular Biophysics, College of Physicians and Surgeons, Columbia University, New York, NY 10032
2 Department of Medicine, College of Physicians and Surgeons, Columbia University, New York, NY 10032

Address correspondence to Samuel C. Silverstein, Dept. of Physiology and Cellular Biophysics, College of Physicians and Surgeons, Columbia University, 630 W. 168th St., New York, NY 10032. Phone: (212) 305-3546; Fax: (212) 305-5775; email: scs3{at}columbia.edu

We showed previously that the competition between bacterial killing by neutrophils and bacterial growth in stirred serum-containing suspensions could be modeled as the competition between a first-order reaction (bacterial growth) and a second-order reaction (bacterial killing by neutrophils). The model provided a useful parameter, the critical neutrophil concentration (CNC), below which bacterial concentration increased and above which it decreased, independent of the initial bacterial concentration. We report here that this model applies to neutrophil killing of bacteria in three-dimensional fibrin matrices and in rabbit dermis. We measured killing of 103–108 colony forming units/ml Staphylococcus epidermidis by 105–108 human neutrophils/ml in fibrin gels. The CNC was ~4 x 106 neutrophils/ml gel in the presence of normal serum and ~1.6 x 107 neutrophils/ml gel in the presence of C5-deficient serum. Application of our model to published data of others on killing of ~5 x 107 to 2 x 108 E. coli/ml rabbit dermis yielded CNCs from ~4 x 106 to ~8 x 106 neutrophils/ml dermis. Thus, in disparate tissues and tissuelike environments, our model fits the kinetics of bacterial killing and gives similar lower limits (CNCs) to the neutrophil concentration required to control bacterial growth.

Key Words: rabbit dermis • bactericidal activity • Staphylococcus epidermidis • fibrin gels • critical neutrophil concentration

Abbreviations used in this paper: CNC, critical neutrophil concentration; NEE, neutrophil extraction efficiency; NS, normal human serum.


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