Different fates ofLegionella pneumophila pmiandmilmutants within macrophages and alveolar epithelial cells

Abstract Alveolar epithelial cells, which constitute the majority of the alveolar surface, may represent a potential niche for intracellular replication ofLegionella pneumophilathat has been largely overlooked. We examined the phenotypes of a bank of 121 macrophage-defective mutants ofL. pneumophila(designated aspmiandmil) for their cytopathogenicity to and intracellular survival and replication within human alveolar epithelial cells. Our data showed that 91 of 121 mutants that were defective (modest–severe) in macrophages exhibited wild type-like phenotypes in human type I alveolar epithelial cells. In contrast, the other 30 mutants were defective in both macrophages and alveolar epithelial cells. Transmission electron microscopy of the intracellular infection by three mutants showed that the defect in intracellular replication in macrophages and epithelial cells was associated with a defect in recruitment of the RER around the phagosome. Differences in attachment to macrophages and epithelial cells were also exhibited by some of the mutants. Pulmonary infection studies of A/J mice showed that a mutant defective in macrophages but not in alveolar epithelial cells replicated like the wild type strain in the lungs of A/J mice. In contrast, a mutant defective in both macrophages and alveolar epithelial cells failed to replicate and was killed. We conclude that certain distinct genetic loci ofL. pneumophilaare uniquely required for intracellular survival and replication within phagocytic but not epithelial cells, which may be important in vivo.

[1]  E R Weibel,et al.  Cell number and cell characteristics of the normal human lung. , 2015, The American review of respiratory disease.

[2]  R. Isberg,et al.  Legionella pneumophila DotA protein is required for early phagosome trafficking decisions that occur within minutes of bacterial uptake , 1998, Molecular microbiology.

[3]  B. Stone,et al.  Expression of Multiple Pili by Legionella pneumophila: Identification and Characterization of a Type IV Pilin Gene and Its Role in Adherence to Mammalian and Protozoan Cells , 1998, Infection and Immunity.

[4]  O. Harb,et al.  Identification of Macrophage-Specific Infectivity Loci (mil) of Legionella pneumophila That Are Not Required for Infectivity of Protozoa , 1998, Infection and Immunity.

[5]  H. Shuman,et al.  Host cell killing and bacterial conjugation require overlapping sets of genes within a 22-kb region of the Legionella pneumophila genome. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[6]  R. Isberg,et al.  Conjugative transfer by the virulence system of Legionella pneumophila. , 1998, Science.

[7]  O. Harb,et al.  Heterogeneity in the Attachment and Uptake Mechanisms of the Legionnaires’ Disease Bacterium, Legionella pneumophila, by Protozoan Hosts , 1998, Applied and Environmental Microbiology.

[8]  R. Marre,et al.  Legionella pneumophila infection in intratracheally inoculated T cell-depleted or -nondepleted A/J mice. , 1998, Journal of immunology.

[9]  Y. Kwaik.  Induced Expression of the Legionella pneumophila Gene Encoding a 20-Kilodalton Protein during Intracellular Infection , 1998, Infection and Immunity.

[10]  O. Harb,et al.  Utilization of similar mechanisms by Legionella pneumophila to parasitize two evolutionarily distant host cells, mammalian macrophages and protozoa , 1997, Infection and immunity.

[11]  Y. Kwaik,et al.  Identification of a Gal/GalNAc Lectin in the Protozoan Hartmannella vermiformis as a Potential Receptor for Attachment and Invasion by the Legionnaires' Disease Bacterium , 1997, The Journal of experimental medicine.

[12]  O. Harb,et al.  Transcriptional regulation of the macrophage‐induced gene (gspA) of Legionella pneumophila and phenotypic characterization of a null mutant , 1997, Molecular microbiology.

[13]  S. Miller,et al.  Rapid and complete fusion of macrophage lysosomes with phagosomes containing Salmonella typhimurium , 1996, Infection and immunity.

[14]  Y. Kwaik,et al.  The use of differential display‐PCR to isolate and characterize a Legionella pneumophila locus induced during the intracellular infection of macrophages , 1996, Molecular microbiology.

[15]  B. Fields,et al.  The molecular ecology of legionellae. , 1996, Trends in microbiology.

[16]  Y. Kwaik.  The phagosome containing Legionella pneumophila within the protozoan Hartmannella vermiformis is surrounded by the rough endoplasmic reticulum. , 1996 .

[17]  J. Hacker,et al.  De novo synthesis of Legionella pneumophila antigens during intracellular growth in phagocytic cells , 1996, Infection and immunity.

[18]  E. Groisman,et al.  Mg2+ as an Extracellular Signal: Environmental Regulation of Salmonella Virulence , 1996, Cell.

[19]  J. Hacker,et al.  Influence of site specifically altered Mip proteins on intracellular survival of Legionella pneumophila in eukaryotic cells , 1995, Infection and immunity.

[20]  B. Finlay,et al.  Targeting of Salmonella typhimurium to vesicles containing lysosomal membrane glycoproteins bypasses compartments with mannose 6-phosphate receptors , 1995, The Journal of cell biology.

[21]  J. Brieland,et al.  Replicative Legionella pneumophila lung infection in intratracheally inoculated A/J mice. A murine model of human Legionnaires' disease. , 1994, The American journal of pathology.

[22]  N. Engleberg,et al.  Cloning and molecular characterization of a Legionella pneumophila gene induced by intracellular infection and by various in vitro stress conditions , 1994, Molecular microbiology.

[23]  N. Engleberg,et al.  Protein expression by the protozoan Hartmannella vermiformis upon contact with its bacterial parasite Legionella pneumophila , 1994, Infection and immunity.

[24]  S. Arata,et al.  Legionella pneumophila growth restriction in permissive macrophages cocultured with nonpermissive lipopolysaccharide-activated macrophages , 1993, Infection and immunity.

[25]  F. Rodgers,et al.  Opsonin-independent adherence and intracellular development of Legionella pneumophila within U-937 cells. , 1993, Canadian journal of microbiology.

[26]  S. Miller,et al.  A PhoP-repressed gene promotes Salmonella typhimurium invasion of epithelial cells , 1993, Journal of bacteriology.

[27]  C. Mody,et al.  Legionella pneumophila replicates within rat alveolar epithelial cells. , 1993, The Journal of infectious diseases.

[28]  N. Engleberg,et al.  Phenotypic modulation by Legionella pneumophila upon infection of macrophages , 1993, Infection and immunity.

[29]  R. Isberg,et al.  Two distinct defects in intracellular growth complemented by a single genetic locus in Legionella pneumophila , 1993, Molecular microbiology.

[30]  W. Johnson,et al.  Adherence of Legionella pneumophila to guinea pig peritoneal macrophages, J774 mouse macrophages, and undifferentiated U937 human monocytes: role of Fc and complement receptors , 1992, Infection and immunity.

[31]  S. Miller,et al.  An unusual pagC::TnphoA mutation leads to an invasion- and virulence-defective phenotype in Salmonellae , 1992, Infection and immunity.

[32]  S. Miller.  PhoP/PhoQ: macrophage‐specific modulators of Salmonella virulence? , 1991, Molecular microbiology.

[33]  C. Mody,et al.  A mutation in the mip gene results in an attenuation of Legionella pneumophila virulence. , 1990, The Journal of infectious diseases.

[34]  L. Mascola,et al.  Role of air sampling in investigation of an outbreak of legionnaires' disease associated with exposure to aerosols from an evaporative condenser. , 1990, The Journal of infectious diseases.

[35]  S. Miller,et al.  Constitutive expression of the phoP regulon attenuates Salmonella virulence and survival within macrophages , 1990, Journal of bacteriology.

[36]  C. Mody,et al.  A Legionella pneumophila gene encoding a species-specific surface protein potentiates initiation of intracellular infection , 1989, Infection and immunity.

[37]  T. Klein,et al.  Role of gamma interferon in induction of natural killer activity by Legionella pneumophila in vitro and in an experimental murine infection model , 1988, Infection and immunity.

[38]  C. Newton,et al.  Growth of Legionella pneumophila in thioglycolate-elicited peritoneal macrophages from A/J mice , 1988, Infection and immunity.

[39]  M. Horwitz,et al.  Interaction between the legionnaires' disease bacterium (Legionella pneumophila) and human alveolar macrophages. Influence of antibody, lymphokines, and hydrocortisone. , 1984, The Journal of clinical investigation.

[40]  M. Horwitz.  The Legionnaires' disease bacterium (Legionella pneumophila) inhibits phagosome-lysosome fusion in human monocytes , 1983, The Journal of experimental medicine.

[41]  M. Horwitz.  Formation of a novel phagosome by the Legionnaires' disease bacterium (Legionella pneumophila) in human monocytes , 1983, The Journal of experimental medicine.

[42]  R. Myerowitz,et al.  The pathology of the Legionella pneumonias. A review of 74 cases and the literature. , 1981, Human pathology.