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Antimicrobial Agents and Chemotherapy, January 2005, p. 269-275, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.269-275.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Antibacterial Activity and Specificity of the Six Human {alpha}-Defensins

Bryan Ericksen,1 Zhibin Wu,1 Wuyuan Lu,1* and Robert I. Lehrer2*

Institute of Human Virology, University of Maryland Biotechnology Institute, Baltimore, Maryland,1 Department of Medicine, David Geffen School of Medicine, University of California, Los Angeles, California2

Received 4 May 2004/ Returned for modification 1 July 2004/ Accepted 10 September 2004

We developed a kinetic, 96-well turbidimetric procedure that is capable of testing the antimicrobial properties of six human {alpha}-defensins concurrently on a single microplate. The defensins were prepared by solid-phase peptide synthesis and tested against gram-positive bacteria (Staphylococcus aureus and Bacillus cereus) and gram-negative bacteria (Enterobacter aerogenes and Escherichia coli). Analysis of the growth curves provided virtual lethal doses (vLDs) equivalent to conventional 50% lethal doses (LD50s), LD90s, LD99s, and LD99.9s obtained from colony counts. On the basis of their respective vLD90s and vLD99s, the relative potencies of human myeloid {alpha}-defensins against S. aureus were HNP2 > HNP1 > HNP3 > HNP4. In contrast, their relative potencies against E. coli and E. aerogenes were HNP4 > HNP2 > HNP1 = HNP3. HD5 was as effective as HNP2 against S. aureus and as effective as HNP4 against the gram-negative bacteria in our panel. HD6 showed little or no activity against any of the bacteria in our panel, including B. cereus, which was highly susceptible to the other five {alpha}-defensins. The assay described provides a quantitative, precise, and economical way to study the antimicrobial activities of host-defense peptides. Its use has clarified the relative potencies of human {alpha}-defensins and raised intriguing questions about the in vivo function(s) of HD6.

* Corresponding author. Mailing address for Wuyuan Lu: Institute of Human Virology, University of Maryland Biotechnology Institute, 725 West Lombard St., Baltimore, MD 21201. Phone: (410) 706-4980. Fax: (410) 706-7583. E-mail: luw{at}umbi.umd.edu. Mailing address for Robert I. Lehrer: Department of Medicine, CHS 37-062, David Geffen School of Medicine, University of California, Los Angeles, CA 90095. Phone: (310) 825-5340. Fax: (310) 206-8766. E-mail: rlehrer{at}mednet.ucla.edu.

Antimicrobial Agents and Chemotherapy, January 2005, p. 269-275, Vol. 49, No. 1
0066-4804/05/$08.00+0     doi:10.1128/AAC.49.1.269-275.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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