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Infection and Immunity, June 2005, p. 3702-3713, Vol. 73, No. 6
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.6.3702-3713.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Opsonized Virulent Brucella abortus Replicates within Nonacidic, Endoplasmic Reticulum-Negative, LAMP-1-Positive Phagosomes in Human Monocytes

Bryan H Bellaire,1* R. Martin Roop II,2 and James A. Cardelli1

Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, Shreveport, Louisiana 71130-3932,1 Depatment of Microbiology and Immunology, East Carolina University School of Medicine, Greenville, North Carolina 278342

Received 31 August 2004/ Returned for modification 16 November 2004/ Accepted 15 February 2005

Cells in the Brucella spp. are intracellular pathogens that survive and replicate within host monocytes. Brucella maintains persistent infections in animals despite the production of high levels of anti-Brucella-specific antibodies. To determine the effect of antibody opsonization on the ability of Brucella to establish itself within monocytes, the intracellular trafficking of virulent Brucella abortus 2308 and attenuated hfq and bacA mutants was followed in the human monocytic cell line THP-1. Early trafficking events of B. abortus 2308-containing phagosomes (BCP) were indistinguishable from those seen for control particles (heat-killed B. abortus 2308, live Escherichia coli HB101, or latex beads). All phagosomes transiently communicated the early-endosomal compartment and rapidly matured into LAMP-1+, cathepsin D+, and acidic phagosomes. By 2 h postinfection, however, the number of cathepsin D+ BCP was significantly lower for live B. abortus 2308-infected cells than for either Brucella mutant strains or control particles. B. abortus 2308 persisted within these cathepsin D–, LAMP-1+, and acidic vesicles; however, at the onset of intracellular replication, the numbers of acidic B. abortus 2308 BCP decreased while remaining cathepsin D– and LAMP-1+. In contrast to B. abortus 2308, the isogenic hfq and bacA mutants remained in acidic, LAMP-1+ phagosomes and failed to initiate intracellular replication. Notably, markers specific for the host endoplasmic reticulum were absent from the BCPs throughout the course of the infection. Thus, opsonized B. abortus in human monocytes survives within phagosomes that remain in the endosomal pathway and replication of virulent B. abortus 2308 within these vesicles corresponds with an increase in intraphagosomal pH.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, Louisiana State University Health Sciences Center, 1501 Kings Highway, Shreveport, LA 71130-3932. Phone: (318) 675-4383. Fax: (318) 675-5764. E-mail: bbella{at}lsuhsc.edu.

Editor: V. J. DiRita

Infection and Immunity, June 2005, p. 3702-3713, Vol. 73, No. 6
0019-9567/05/$08.00+0     doi:10.1128/IAI.73.6.3702-3713.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



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