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Infection and Immunity, May 2000, p. 2939-2947, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
Intracellular Growth of Legionella
pneumophila in Dictyostelium discoideum, a System for
Genetic Analysis of Host-Pathogen Interactions
Jonathan M.
Solomon,1
Adam
Rupper,2
James A.
Cardelli,2 and
Ralph
R.
Isberg1,*
Howard Hughes Medical Institute, Department
of Molecular Biology and Microbiology, Tufts University
Medical School, Boston, Massachusetts
02111,1 and Department of Microbiology
and Immunology LSU Health Sciences Center and Feist-Weiller
Cancer Center, Shreveport, Louisiana 711302
Received 6 December 1999/Accepted 8 February 2000
Conditions were established in which Legionella
pneumophila, an intracellular bacterial pathogen, could replicate
within the unicellular organism Dictyostelium discoideum.
By several criteria, L. pneumophila grew by the same
mechanism within D. discoideum as it does in amoebae and
macrophages. Bacteria grew within membrane-bound vesicles associated
with rough endoplasmic reticulum, and L. pneumophila dot/icm mutants, blocked for growth in macrophages and amoebae, also did not grow in D. discoideum. Internalized L. pneumophila avoided degradation by D. discoideum and
showed evidence of reduced fusion with endocytic compartments. The
ability of L. pneumophila to grow within D. discoideum depended on the growth state of the cells. D. discoideum grown as adherent monolayers was susceptible to
L. pneumophila infection and to contact-dependent
cytotoxicity during high-multiplicity infections, whereas D. discoideum grown in suspension was relatively resistant to
cytotoxicity and did not support intracellular growth. Some known
D. discoideum mutants were examined for their effect on
growth of L. pneumophila. The coronin mutant and the
myoA/B double myosin I mutant were more permissive than
wild-type strains for intracellular growth. Growth of L. pneumophila in a G
mutant was slightly reduced
compared to the parent strain. This work demonstrates the usefulness of the L. pneumophila-D. discoideum system for genetic
analysis of host-pathogen interactions.
*
Corresponding author. Mailing address: Howard Hughes
Medical Institute, Department of Molecular Biology and Microbiology, Tufts University Medical School, 136 Harrison Ave., Boston, MA 02111. Phone: (617) 636 3993. Fax: (617) 636 0337. E-mail:
risberg{at}opal.tufts.edu.
Infection and Immunity, May 2000, p. 2939-2947, Vol. 68, No. 5
0019-9567/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.
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