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Infection and Immunity, August 1999, p. 4041-4047, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Early Acidification of Phagosomes Containing
Brucella suis Is Essential for Intracellular Survival in
Murine Macrophages
Françoise
Porte,*
Jean-Pierre
Liautard, and
Stephan
Köhler
Institut National de la Santé et de la
Recherche Médicale U-431, Montpellier, France
Received 28 December 1998/Returned for modification 9 March
1999/Accepted 4 May 1999
Brucella suis is a facultative intracellular pathogen
of mammals, residing in macrophage vacuoles. In this work, we studied the phagosomal environment of these bacteria in order to better understand the mechanisms allowing survival and multiplication of
B. suis. Intraphagosomal pH in murine J774 cells was
determined by measuring the fluorescence intensity of opsonized,
carboxyfluorescein-rhodamine- and Oregon Green 488-rhodamine-labeled
bacteria. Compartments containing live B. suis acidified to
a pH of about 4.0 to 4.5 within 60 min. Acidification of B. suis-containing phagosomes in the early phase of infection was
abolished by treatment of host cells with 100 nM bafilomycin
A1, a specific inhibitor of vacuolar proton-ATPases. This
neutralization at 1 h postinfection resulted in a 2- to 34-fold
reduction of opsonized and nonopsonized viable intracellular bacteria
at 4 and 6 h postinfection, respectively. Ammonium chloride and
monensin, other pH-neutralizing reagents, led to comparable loss of
intracellular viability. Addition of ammonium chloride at 7 h
after the beginning of infection, however, did not affect intracellular
multiplication of B. suis, in contrast to treatment at
1 h postinfection, where bacteria were completely eradicated
within 48 h. Thus, we conclude that phagosomes with B. suis acidify rapidly after infection, and that this early
acidification is essential for replication of the bacteria within the macrophage.
*
Corresponding author. Mailing address: INSERM U-431,
Université Montpellier II, C.P. 100, Pl. E. Bataillon, 34095 Montpellier, France. Phone: (33) 4 67 14 42 38. Fax: (33) 4 67 14 33 38. E-mail: porte{at}crit.univ-montp2.fr.
Infection and Immunity, August 1999, p. 4041-4047, Vol. 67, No. 8
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
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