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Infect. Immun., Mar 1997, 1023-1031, Vol 65, No. 3
MR Yeaman, YQ Tang, AJ Shen, AS Bayer and ME Selsted
Recent in vitro studies have demonstrated that rabbit platelets release a
small, cationic antimicrobial protein in response to thrombin stimulation
under physiological conditions (M. R. Yeaman, S. M. Puentes, D. C. Norman,
and A. S. Bayer, Infect. Immun. 60:1202-1209, 1992). This observation
prompted our present investigation, focused on determining the array of
antimicrobial proteins contained within rabbit platelets and their in vitro
activity against common bloodstream pathogens. A group of small (6.0- to
9.0-kDa), cationic proteins with in vitro antimicrobial activity was
purified from whole and thrombin- stimulated rabbit platelets by gel
filtration and reversed-phase high- performance liquid chromatography.
Purified proteins in micromolar concentrations (10 to 40 microg/ml) exerted
in vitro microbiostatic and/or microbicidal activities against
Staphylococcus aureus, Escherichia coli, and Candida albicans in a
dose-dependent manner. The antimicrobial activities of proteins purified
from rabbit platelet acid extracts were generally inversely related to pH,
with maximal activity observed at pH 5.5. In contrast, the predominant
protein isolated from thrombin-stimulated rabbit platelets, though
biochemically and microbiologically similar to proteins extracted by acid,
exhibited antimicrobial activities which were modestly enhanced at pH 7.2
compared with pH 5.5. Amino acid compositional analyses in combination with
molecular mass determinations suggest that the majority of these proteins
are distinct molecules not derived from a single common precursor.
Collectively, these data indicate that rabbit platelets contain proteins
which exert potent in vitro antimicrobial activity against bacterial and
fungal pathogens which commonly invade the bloodstream. Moreover, several
of these proteins were released from platelets stimulated with thrombin
under physiological conditions and exerted potent antimicrobial activities
in physiological pH ranges. These observations support the hypothesis that
platelets serve an important role in host defense against infection, via
localized release of antimicrobial proteins in response to stimuli
associated with tissue injury or microbial colonization.
Copyright © 1997, American Society for Microbiology
Purification and in vitro activities of rabbit platelet microbicidal proteins
Department of Medicine, Los Angeles County-Harbor UCLA Medical Center, Torrance, California 90509, USA. yeaman@afp76.humc.edu
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