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Infect Immun, April 1998, p. 1718-1725, Vol. 66, No. 4
Laboratoire des
Bordetella1 and
Unité de
Pharmacologie Cellulaire, INSERM U485,2 Institut
Pasteur, Paris, France
Received 9 October 1997/Returned for modification 18 November
1997/Accepted 17 December 1997
Bordetella pertussis induces in vitro apoptosis of
murine alveolar macrophages by a mechanism that is dependent on
expression of bacterial adenylate cyclase-hemolysin. Using a murine
respiratory model, we found in this study that intranasal infection
with a parental B. pertussis strain, but not with an
isogenic variant deficient in the expression of all toxins and
adhesins, induced a marked neutrophil accumulation in the
bronchoalveolar lavage fluid and an early decrease in macrophage
numbers. These phenomena paralleled a time-dependent rise in the
proportion of apoptotic nuclei, as detected by flow cytometry, and of
macrophages which had engulfed apoptotic bodies. Apoptotic death of
bronchopulmonary cells was observed exclusively following intranasal
infection with bacteria reisolated from lungs of infected animals and
not with B. pertussis collected after in vitro subculture.
Using the terminal deoxynucleotidyltransferase-mediated dUTP-biotin
nick end labeling technique coupled to fluorescence microscopy and morphological analysis, we established that the apoptotic cells in
bronchoalveolar lavage fluids were neutrophils and macrophages. Histological analysis of the lung tissues from B. pertussis-infected mice showed increased numbers of apoptotic
cells in the alveolar compartments. Cellular accumulation in
bronchoalveolar lavage fluids and apoptosis of alveolar macrophages
were significantly attenuated in mice infected with a mutant deficient
in the expression of adenylate cyclase-hemolysin, indicating a role of
this enzyme in these processes.
0019-9567/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.
Role of Adenylate Cyclase-Hemolysin in Alveolar
Macrophage Apoptosis during Bordetella pertussis Infection
In Vivo
*
Corresponding author. Mailing address: Laboratoire des
Bordetella, Centre National de Référence des Bordetelles,
Institut Pasteur, 28, rue du Dr. Roux, 75724 Paris Cedex 15, France.
Phone: (33-1) 45.68.83.34. Fax: (33-1) 40.61.35.33. E-mail:
nguiso{at}pasteur.fr.
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