16 results on '"Coleman FT"'
Search Results
2. Stimulation of a subset of natural killer T cells by CD103 + DC is required for GM-CSF and protection from pneumococcal infection.
- Author
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Murray MP, Crosby CM, Marcovecchio P, Hartmann N, Chandra S, Zhao M, Khurana A, Zahner SP, Clausen BE, Coleman FT, Mizgerd JP, Mikulski Z, and Kronenberg M
- Subjects
- Animals, Cell Line, Dendritic Cells immunology, Female, Humans, Interferon-gamma immunology, Interleukin-17 immunology, Intraepithelial Lymphocytes immunology, Intraepithelial Lymphocytes metabolism, Lung immunology, Lung microbiology, Lymphocyte Activation, Male, Mice, Mice, Inbred C57BL, Pneumococcal Infections immunology, Receptors, Antigen, T-Cell, gamma-delta immunology, Streptococcus pneumoniae immunology, Granulocyte-Macrophage Colony-Stimulating Factor metabolism, Natural Killer T-Cells immunology, Natural Killer T-Cells metabolism
- Abstract
Innate-like T cells, including invariant natural killer T cells, mucosal-associated invariant T cells, and γδ T cells, are present in various barrier tissues, including the lung, where they carry out protective responses during infections. Here, we investigate their roles during pulmonary pneumococcal infection. Following infection, innate-like T cells rapidly increase in lung tissue, in part through recruitment, but T cell antigen receptor activation and cytokine production occur mostly in interleukin-17-producing NKT17 and γδ T cells. NKT17 cells are preferentially located within lung tissue prior to infection, as are CD103
+ dendritic cells, which are important both for antigen presentation to NKT17 cells and γδ T cell activation. Whereas interleukin-17-producing γδ T cells are numerous, granulocyte-macrophage colony-stimulating factor is exclusive to NKT17 cells and is required for optimal protection. These studies demonstrate how particular cellular interactions and responses of functional subsets of innate-like T cells contribute to protection from pathogenic lung infection., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)- Published
- 2022
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3. Riboflavin Metabolism Variation among Clinical Isolates of Streptococcus pneumoniae Results in Differential Activation of Mucosal-associated Invariant T Cells.
- Author
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Hartmann N, McMurtrey C, Sorensen ML, Huber ME, Kurapova R, Coleman FT, Mizgerd JP, Hildebrand W, Kronenberg M, Lewinsohn DM, and Harriff MJ
- Subjects
- Animals, Cytokines metabolism, Dendritic Cells microbiology, Gene Expression Regulation, Bacterial, Humans, Mice, Mutant Strains, Phagocytosis, Respiratory Mucosa microbiology, Riboflavin genetics, Streptococcus pneumoniae isolation & purification, Streptococcus pneumoniae pathogenicity, Host-Pathogen Interactions physiology, Respiratory Mucosa cytology, Riboflavin metabolism, Streptococcus pneumoniae metabolism, T-Lymphocytes microbiology
- Abstract
Streptococcus pneumoniae is an important bacterial pathogen that causes a range of noninvasive and invasive diseases. The mechanisms underlying variability in the ability of S. pneumoniae to transition from nasopharyngeal colonization to disease-causing pathogen are not well defined. Mucosal-associated invariant T (MAIT) cells are prevalent in mucosal tissues such as the airways and are believed to play an important role in the early response to infection with bacterial pathogens. The ability of MAIT cells to recognize and contain infection with S. pneumoniae is not known. In the present study, we analyzed MAIT-cell responses to infection with clinical isolates of S. pneumoniae serotype 19A, a serotype linked to invasive pneumococcal disease. We found that although MAIT cells were capable of responding to human dendritic and airway epithelial cells infected with S. pneumoniae, the magnitude of response to different serotype 19A isolates was determined by genetic differences in the expression of the riboflavin biosynthesis pathway. MAIT-cell release of cytokines correlated with differences in the ability of MAIT cells to respond to and control S. pneumoniae in vitro and in vivo in a mouse challenge model. Together, these results demonstrate first that there are genetic differences in riboflavin metabolism among clinical isolates of the same serotype and second that these likely determine MAIT-cell function in response to infection with S. pneumoniae. These differences are critical when considering the role that MAIT cells play in early responses to pneumococcal infection and determining whether invasive disease will develop.
- Published
- 2018
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4. Regionally compartmentalized resident memory T cells mediate naturally acquired protection against pneumococcal pneumonia.
- Author
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Smith NM, Wasserman GA, Coleman FT, Hilliard KL, Yamamoto K, Lipsitz E, Malley R, Dooms H, Jones MR, Quinton LJ, and Mizgerd JP
- Subjects
- Animals, Bacterial Load, Cellular Microenvironment, Female, Immunologic Memory, Interleukin-17 metabolism, Lung microbiology, Mice, Mice, Inbred C57BL, Pneumonia, Pneumococcal microbiology, Serogroup, Virulence, CD4-Positive T-Lymphocytes immunology, Immunity, Heterologous, Lung immunology, Pneumonia, Pneumococcal immunology, Streptococcus pneumoniae physiology
- Abstract
As children age, they become less susceptible to the diverse microbes causing pneumonia. These microbes are pathobionts that infect the respiratory tract multiple times during childhood, generating immunological memory. To elucidate mechanisms of such naturally acquired immune protection against pneumonia, we modeled a relevant immunological history in mice by infecting their airways with mismatched serotypes of Streptococcus pneumoniae (pneumococcus). Previous pneumococcal infections provided protection against a heterotypic, highly virulent pneumococcus, as evidenced by reduced bacterial burdens and long-term sterilizing immunity. This protection was diminished by depletion of CD4
+ cells prior to the final infection. The resolution of previous pneumococcal infections seeded the lungs with CD4+ resident memory T (TRM ) cells, which responded to heterotypic pneumococcus stimulation by producing multiple effector cytokines, particularly interleukin (IL)-17A. Following lobar pneumonias, IL-17-producing CD4+ TRM cells were confined to the previously infected lobe, rather than dispersed throughout the lower respiratory tract. Importantly, pneumonia protection also was confined to that immunologically experienced lobe. Thus regionally localized memory cells provide superior local tissue protection to that mediated by systemic or central memory immune defenses. We conclude that respiratory bacterial infections elicit CD4+ TRM cells that fill a local niche to optimize heterotypic protection of the affected tissue, preventing pneumonia.- Published
- 2018
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5. Capacity of Pneumococci to Activate Macrophage Nuclear Factor κB: Influence on Necroptosis and Pneumonia Severity.
- Author
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Coleman FT, Blahna MT, Kamata H, Yamamoto K, Zabinski MC, Kramnik I, Wilson AA, Kotton DN, Quinton LJ, Jones MR, Pelton SI, and Mizgerd JP
- Subjects
- Animals, Disease Models, Animal, Female, Immunity, Innate, Macrophages, Alveolar microbiology, Male, Mice, Mice, Inbred C57BL, Necrosis therapy, Pneumonia, Pneumococcal therapy, RAW 264.7 Cells, Reactive Oxygen Species metabolism, Macrophages, Alveolar metabolism, NF-kappa B metabolism, Necrosis immunology, Pneumonia, Pneumococcal immunology, Streptococcus pneumoniae
- Abstract
During pneumococcal pneumonia, antibacterial defense requires the orchestrated expression of innate immunity mediators, initiated by alveolar macrophages and dependent on transcription driven by nuclear factor κB (NF-κB). Such immune pressure may select for pneumococci, which avoid or subvert macrophage NF-κB activation. Analyzing pneumococci collected from children in Massachusetts, we found that the activation of macrophage NF-κB by Streptococcus pneumoniae is highly diverse, with a preponderance of low NF-κB activators that associate particularly with complicated pneumonia. Low NF-κB activators cause more severe lung infections in mice, and they drive macrophages toward an alternate and detrimental cell fate of necroptosis. Both outcomes can be reversed by activation of macrophages with pneumococci that are high NF-κB activators. These results suggest that low NF-κB activation is a virulence property of pneumococci and that the appropriate activation of macrophages, including NF-κB, may hold promise as an adjunct therapeutic avenue for pneumococcal pneumonia., (© The Author 2017. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)
- Published
- 2017
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6. Myeloid ZFP36L1 does not regulate inflammation or host defense in mouse models of acute bacterial infection.
- Author
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Hyatt LD, Wasserman GA, Rah YJ, Matsuura KY, Coleman FT, Hilliard KL, Pepper-Cunningham ZA, Ieong M, Stumpo DJ, Blackshear PJ, Quinton LJ, Mizgerd JP, and Jones MR
- Subjects
- Animals, Bacterial Infections microbiology, Bronchoalveolar Lavage Fluid microbiology, Butyrate Response Factor 1, Cell Line, Cytokines metabolism, Humans, Inflammation microbiology, Lung metabolism, Lung microbiology, Macrophages metabolism, Macrophages microbiology, Mice, Mice, Inbred C57BL, Myeloid Cells metabolism, Myeloid Cells microbiology, Pneumonia, Bacterial microbiology, RNA, Messenger metabolism, Bacterial Infections metabolism, Inflammation metabolism, Nuclear Proteins metabolism, Pneumonia, Bacterial metabolism, RNA-Binding Proteins metabolism
- Abstract
Zinc finger protein 36, C3H type-like 1 (ZFP36L1) is one of several Zinc Finger Protein 36 (Zfp36) family members, which bind AU rich elements within 3' untranslated regions (UTRs) to negatively regulate the post-transcriptional expression of targeted mRNAs. The prototypical member of the family, Tristetraprolin (TTP or ZFP36), has been well-studied in the context of inflammation and plays an important role in repressing pro-inflammatory transcripts such as TNF-α. Much less is known about the other family members, and none have been studied in the context of infection. Using macrophage cell lines and primary alveolar macrophages we demonstrated that, like ZFP36, ZFP36L1 is prominently induced by infection. To test our hypothesis that macrophage production of ZFP36L1 is necessary for regulation of the inflammatory response of the lung during pneumonia, we generated mice with a myeloid-specific deficiency of ZFP36L1. Surprisingly, we found that myeloid deficiency of ZFP36L1 did not result in alteration of lung cytokine production after infection, altered clearance of bacteria, or increased inflammatory lung injury. Although alveolar macrophages are critical components of the innate defense against respiratory pathogens, we concluded that myeloid ZFP36L1 is not essential for appropriate responses to bacteria in the lungs. Based on studies conducted with myeloid-deficient ZFP36 mice, our data indicate that, of the Zfp36 family, ZFP36 is the predominant negative regulator of cytokine expression in macrophages. In conclusion, these results imply that myeloid ZFP36 may fully compensate for loss of ZFP36L1 or that Zfp36l1-dependent mRNA expression does not play an integral role in the host defense against bacterial pneumonia.
- Published
- 2014
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7. Resistance to Pseudomonas aeruginosa chronic lung infection requires cystic fibrosis transmembrane conductance regulator-modulated interleukin-1 (IL-1) release and signaling through the IL-1 receptor.
- Author
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Reiniger N, Lee MM, Coleman FT, Ray C, Golan DE, and Pier GB
- Subjects
- Animals, Cell Line, Cell Nucleus metabolism, Cystic Fibrosis Transmembrane Conductance Regulator genetics, Diptera, Humans, Immunity, Innate, Interleukin-1beta metabolism, Lung immunology, Lung metabolism, Mice, Mice, Inbred Strains, Mice, Knockout, Microscopy, Confocal, Myeloid Differentiation Factor 88 genetics, Myeloid Differentiation Factor 88 immunology, NF-kappa B metabolism, Pseudomonas Infections metabolism, Receptors, Interleukin-1 genetics, Toll-Like Receptor 2 immunology, Toll-Like Receptor 4 immunology, Toll-Like Receptor 5 immunology, Cystic Fibrosis Transmembrane Conductance Regulator physiology, Interleukin-1beta immunology, Pseudomonas Infections immunology, Pseudomonas aeruginosa immunology, Receptors, Interleukin-1 immunology
- Abstract
Innate immunity is critical for clearing Pseudomonas aeruginosa from the lungs. In response to P. aeruginosa infection, a central transcriptional regulator of innate immunity-NF-kappaB-is translocated within 15 min to the nuclei of respiratory epithelial cells expressing wild-type (WT) cystic fibrosis (CF) transmembrane conductance regulator (CFTR). P. aeruginosa clearance from lungs is impaired in CF, and rapid NF-kappaB nuclear translocation is defective in cells with mutant or missing CFTR. We used WT and mutant P. aeruginosa and strains of transgenic mice lacking molecules involved in innate immunity to identify additional mediators required for P. aeruginosa-induced rapid NF-kappaB nuclear translocation in lung epithelia. We found neither Toll-like receptor 2 (TLR2) nor TLR4 nor TLR5 were required for this response. However, both MyD88-deficient mice and interleukin-1 receptor (IL-1R)-deficient mice failed to rapidly translocate NF-kappaB to the nuclei of respiratory epithelial cells in response to P. aeruginosa. Cultured human bronchial epithelial cells rapidly released IL-1beta in response to P. aeruginosa; this process was maximized by expression of WT-CFTR and dramatically muted in cells with DeltaF508-CFTR. The IL-1R antagonist blocked P. aeruginosa-induced NF-kappaB nuclear translocation. Oral inoculation via drinking water of IL-1R knockout mice resulted in higher rates of lung colonization and elevated P. aeruginosa-specific antibody titers in a manner analogous to that of CFTR-deficient mice. Overall, rapid IL-1 release and signaling through IL-1R represent key steps in the innate immune response to P. aeruginosa infection, and this process is deficient in cells lacking functional CFTR.
- Published
- 2007
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8. Human monoclonal antibodies to Pseudomonas aeruginosa alginate that protect against infection by both mucoid and nonmucoid strains.
- Author
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Pier GB, Boyer D, Preston M, Coleman FT, Llosa N, Mueschenborn-Koglin S, Theilacker C, Goldenberg H, Uchin J, Priebe GP, Grout M, Posner M, and Cavacini L
- Subjects
- Acute Disease, Alginates metabolism, Animals, Antibodies, Bacterial biosynthesis, Antibodies, Bacterial genetics, Antibodies, Bacterial metabolism, Antibodies, Monoclonal biosynthesis, Antibodies, Monoclonal genetics, Antibodies, Monoclonal metabolism, Binding Sites, Antibody, Disease Models, Animal, Female, Glucuronic Acid metabolism, Hexuronic Acids metabolism, Humans, Hybridomas, Immunoglobulin Variable Region administration & dosage, Immunoglobulin Variable Region biosynthesis, Immunoglobulin Variable Region genetics, Immunoglobulin Variable Region metabolism, Lung immunology, Lung microbiology, Mice, Mice, Inbred C3H, Mice, Inbred C57BL, Phagocytosis immunology, Pneumonia, Bacterial immunology, Pneumonia, Bacterial microbiology, Pneumonia, Bacterial prevention & control, Pseudomonas Infections microbiology, Pseudomonas aeruginosa pathogenicity, Recombinant Proteins administration & dosage, Recombinant Proteins biosynthesis, Recombinant Proteins metabolism, Species Specificity, Antibodies, Bacterial administration & dosage, Antibodies, Monoclonal administration & dosage, Glucuronic Acid immunology, Hexuronic Acids immunology, Pseudomonas Infections immunology, Pseudomonas Infections prevention & control, Pseudomonas aeruginosa immunology
- Abstract
Two fully human mAbs specific for epitopes dependent on intact carboxylate groups on the C6 carbon of the mannuronic acid components of Pseudomonas aeruginosa alginate were found to promote phagocytic killing of both mucoid and nonmucoid strains as well as protection against both types of strains in a mouse model of acute pneumonia. The specificity of the mAbs for alginate was determined by ELISA and killing assays. Some strains of P. aeruginosa did not make detectable alginate in vitro, but in vivo protection against lethal pneumonia was obtained and shown to be due to rapid induction of expression of alginate in the murine lung. No protection against strains genetically unable to make alginate was achieved. These mAbs have potential to be passive therapeutic reagents for all strains of P. aeruginosa and the results document that alginate is a target for the proper type of protective Ab even when expressed at low levels on phenotypically nonmucoid strains.
- Published
- 2004
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9. The galU Gene of Pseudomonas aeruginosa is required for corneal infection and efficient systemic spread following pneumonia but not for infection confined to the lung.
- Author
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Priebe GP, Dean CR, Zaidi T, Meluleni GJ, Coleman FT, Coutinho YS, Noto MJ, Urban TA, Pier GB, and Goldberg JB
- Subjects
- Animals, Female, Lung metabolism, Lung pathology, Mice, Mutation, Pneumonia metabolism, Pneumonia pathology, Pseudomonas aeruginosa enzymology, Pseudomonas aeruginosa immunology, Pseudomonas aeruginosa pathogenicity, Virulence, Cornea microbiology, Eye Infections, Bacterial enzymology, Pseudomonas aeruginosa genetics, UTP-Glucose-1-Phosphate Uridylyltransferase genetics
- Abstract
Acute pneumonias and corneal infections due to Pseudomonas aeruginosa are typically caused by lipopolysaccharide (LPS)-smooth strains. In cystic fibrosis patients, however, LPS-rough strains of P. aeruginosa, which lack O antigen, can survive in the lung and cause chronic infection. It is not clear whether an LPS-rough phenotype affects cytotoxicity related to the type III secretion system (TTSS). We previously reported that interruption of the galU gene in P. aeruginosa results in production of a rough LPS and truncated LPS core. Here we evaluated the role of the galU gene in the pathogenesis of murine lung and eye infections and in cytotoxicity due to the TTSS effector ExoU. We studied galU mutants of strain PAO1, of its cytotoxic variant expressing ExoU from a plasmid, and of the inherently cytotoxic strain PA103. The galU mutants were more serum sensitive than the parental strains but remained cytotoxic in vitro. In a corneal infection model, the galU mutants were significantly attenuated. In an acute pneumonia model, the 50% lethal doses of the galU mutants were higher than those of the corresponding wild-type strains, yet these mutants could cause mortality and severe pneumonia, as judged by histology, even with minimal systemic spread. These findings suggest that the galU gene is required for corneal infection and for efficient systemic spread following lung infection but is not required for infection confined to the lung. Host defenses in the lung appear to be insufficient to control infection with LPS-rough P. aeruginosa when local bacterial levels are high.
- Published
- 2004
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10. Construction and characterization of a Pseudomonas aeruginosa mucoid exopolysaccharide-alginate conjugate vaccine.
- Author
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Theilacker C, Coleman FT, Mueschenborn S, Llosa N, Grout M, and Pier GB
- Subjects
- Animals, Antibodies, Bacterial blood, Blood Bactericidal Activity, Enzyme-Linked Immunosorbent Assay, Epitopes, Hemocyanins immunology, Mice, Mice, Inbred C3H, Rabbits, Vaccines, Conjugate immunology, Alginates chemistry, Bacterial Vaccines immunology, Polysaccharides, Bacterial immunology, Pseudomonas aeruginosa immunology
- Abstract
Deterioration of lung function in patients with cystic fibrosis (CF) is closely associated with chronic pulmonary infection with mucoid Pseudomonas aeruginosa. The mucoid exopolysaccharide (MEP) from P. aeruginosa has been shown to induce opsonic antibodies in mice that are protective against this chronic infection. MEP-specific opsonic antibodies are also commonly found in the sera of older CF patients lacking detectable P. aeruginosa infection. When used in a human vaccine trial, however, MEP only minimally induced opsonic antibodies. To evaluate whether conjugation of MEP to a carrier protein could improve its immunogenicity, we bound thiolated MEP to keyhole limpet hemocyanin (KLH) by using succinimidyl-4-(N-maleimidomethyl)cyclohexane-1-carboxylate (SMCC) as a linker. In contrast to the native MEP polymer, the MEP-KLH conjugate vaccine induced high titers of MEP-specific immunoglobulin G (IgG) in C3H-HeN mice and in a rabbit. Sera from mice immunized with MEP-KLH conjugate, but not from animals immunized with comparable doses of native MEP, demonstrated opsonic killing activity. Vaccination with MEP-KLH conjugate induced opsonic antibodies broadly cross-reactive to heterologous mucoid strains of P. aeruginosa. Preexisting nonopsonic antibodies to MEP are found in normal human sera, including young CF patients, and their presence impedes the induction of opsonic antibodies. Induction of nonopsonic antibodies by either intraperitoneal injection of MEP or injection or feeding of the cross-reactive antigen, seaweed alginate, reduced the level of overall IgG elicited by follow-up immunization with the MEP-KLH conjugate. However, the opsonic activity was lower only in the sera of MEP-KLH conjugate-immunized mice with preexisting antibodies induced by MEP but not with antibodies induced by seaweed alginate. Immunization with MEP-KLH elicited a significant proportion of antibodies specific to epitopes involving O-acetate residues, and this subpopulation of antibodies mediated opsonic killing of mucoid P. aeruginosa in vitro. These results indicate that conjugation of MEP to KLH significantly enhances its immunogenicity and the elicitation of opsonic antibodies in mice and rabbits, that the conjugate induces opsonic antibodies in the presence of preexisting nonopsonic antibodies, and that opsonic antibodies to MEP are directed at epitopes that include acetate residues on the uronic acid polymer.
- Published
- 2003
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11. Protection against fatal Pseudomonas aeruginosa pneumonia in mice after nasal immunization with a live, attenuated aroA deletion mutant.
- Author
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Priebe GP, Meluleni GJ, Coleman FT, Goldberg JB, and Pier GB
- Subjects
- Administration, Intranasal, Animals, Female, Immunization, Lethal Dose 50, Lipopolysaccharides immunology, Lung pathology, Mice, Mice, Inbred Strains, O Antigens genetics, Rabbits, Vaccines, Attenuated immunology, Bacterial Vaccines immunology, O Antigens physiology, Pneumonia, Bacterial prevention & control, Pseudomonas Infections prevention & control
- Abstract
Studies of immunity to Pseudomonas aeruginosa have indicated that a variety of potential immunogens can elicit protection in animal models, utilizing both antibody- and cell-mediated immune effectors for protection. To attempt to optimize delivery of multiple protective antigens and elicit a broad range of immune effectors, we produced an aroA deletion mutant of the P. aeruginosa serogroup O2/O5 strain PAO1, designated PAO1deltaaroA. Previously, we reported that this strain elicits high levels of opsonic antibody directed against many serogroup O2/O5 strains after nasal immunization of mice and rabbits. Here, we assessed the protective efficacy of immunization with PAO1deltaaroA against acute fatal pneumonia in mice. After active immunization, high levels of protection were achieved against an ExoU-expressing cytotoxic variant of the parental strain PAO1 at doses up to 1,000-fold greater than the 50% lethal dose. Significant protection against PAO1 and two of four other serogroup O2/O5 strains was also found, but there was no protection against serogroup-heterologous strains. The serogroup O2/O5 strains not protected against were killed in opsonophagocytic assays as efficiently as the strains with which protection was seen, indicating a lack of correlation of protection and opsonic killing within the serogroup. In passive immunization experiments using challenge with wild-type PAO1 or other noncytotoxic members of the O2/O5 serogroup, there was no protection despite the presence of high levels of opsonic antibody in the mouse sera. However, passive immunization did prevent mortality from pneumonia due to the cytotoxic PAO1 variant at low-challenge doses. These data suggest that a combination of humoral and cellular immunity is required for protection against P. aeruginosa lung infections, that such immunity can be elicited by using aroA deletion mutants, and that a multivalent P. aeruginosa vaccine composed of aroA deletion mutants of multiple serogroups holds significant promise.
- Published
- 2003
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12. Hypersusceptibility of cystic fibrosis mice to chronic Pseudomonas aeruginosa oropharyngeal colonization and lung infection.
- Author
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Coleman FT, Mueschenborn S, Meluleni G, Ray C, Carey VJ, Vargas SO, Cannon CL, Ausubel FM, and Pier GB
- Subjects
- Animals, Chronic Disease, Cystic Fibrosis microbiology, Cystic Fibrosis pathology, Mice, Pseudomonas Infections microbiology, Pseudomonas Infections pathology, Pseudomonas aeruginosa pathogenicity, Virulence, Cystic Fibrosis complications, Mouth microbiology, Pharynx microbiology, Pneumonia, Bacterial complications, Pseudomonas Infections complications, Pseudomonas aeruginosa isolation & purification
- Abstract
No transgenic cystic fibrosis (CF) mouse model developed to date mimics the major clinical phenotype found in humans with CF, chronic Pseudomonas aeruginosa lung infection. In a transgenic CF transmembrane conductance regulator (cftr) mouse colony, we found WT, heterozygous, and homozygous CF mice housed in the same cage became chronically colonized in the oropharynx with environmental P. aeruginosa when the bacterium was present in drinking water. Elimination of P. aeruginosa from drinking water resulted in clearance in most WT and CF heterozygous, but not homozygous mice. For experimental evaluation, a combination of specific animal husbandry techniques and an oral infection route showed cftr(-/-) mice but not WT mice can be chronically colonized by P. aeruginosa with subsequent lung translocation, yielding a pathologic picture indicative of chronic lung infection. In some instances, mucoid isolates of P. aeruginosa were recovered from lungs, indicating conditions were present for conversion to mucoidy. Overexpression of human CFTR in the lungs of WT mice markedly accelerated the clearance rate of P. aeruginosa, demonstrating that lung levels of CFTR play an important role in defense against infection. P. aeruginosa mutants unable to express the surface polysaccharide alginate or the global regulator GacA were deficient in their ability to colonize the mice. CF mice made potent immune responses to P. aeruginosa outer membrane antigens. Overall, we found that under the proper conditions, transgenic CF mice are hypersusceptible to P. aeruginosa colonization and infection and can be used for evaluations of lung pathophysiology, bacterial virulence, and development of therapies aimed at treating CF lung disease.
- Published
- 2003
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13. Construction and characterization of a live, attenuated aroA deletion mutant of Pseudomonas aeruginosa as a candidate intranasal vaccine.
- Author
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Priebe GP, Brinig MM, Hatano K, Grout M, Coleman FT, Pier GB, and Goldberg JB
- Subjects
- 3-Phosphoshikimate 1-Carboxyvinyltransferase, Administration, Intranasal, Animals, Antibodies, Bacterial blood, Bacterial Vaccines administration & dosage, Gene Deletion, Mice, Mice, Inbred BALB C, Mutation, Phagocytosis, Pseudomonas aeruginosa genetics, Pseudomonas aeruginosa pathogenicity, Rabbits, Vaccines, Attenuated, Vaccines, Synthetic administration & dosage, Vaccines, Synthetic therapeutic use, Alkyl and Aryl Transferases genetics, Bacterial Vaccines therapeutic use, Pseudomonas Infections prevention & control, Pseudomonas aeruginosa immunology, Vaccination
- Abstract
Antibodies to the lipopolysaccharide O antigen of Pseudomonas aeruginosa mediate high-level immunity, but protective epitopes have proven to be poorly immunogenic, while nonprotective or minimally protective O-antigen epitopes often elicit the best immune responses. With the goal of developing a broadly protective P. aeruginosa vaccine, we used a gene replacement system based on the Flp recombinase to construct an unmarked aroA deletion mutant of the P. aeruginosa serogroup O2/O5 strain PAO1. The resultant aroA deletion mutant of PAO1 is designated PAO1 Delta aroA. The aroA deletion was confirmed by both PCR and failure of the mutant to grow on minimal media lacking aromatic amino acids. When evaluated for safety and immunogenicity in mice, PAO1 Delta aroA could be applied either intranasally or intraperitoneally at doses up to 5 x 10(9) CFU per mouse without adverse effects. No dissemination of PAO1 Delta aroA to blood, liver, or spleen was detected after intranasal application, and histological evidence of pneumonia was minimal. Intranasal immunization of mice and rabbits elicited high titers of immunoglobulin G to whole bacterial cells and to heat-stable bacterial antigens of all seven prototypic P. aeruginosa serogroup O2/O5 strains. The mouse antisera mediated potent phagocytic killing of most of the prototypic serogroup O2/O5 strains, while the rabbit antisera mediated phagocytic killing of several serogroup-heterologous strains in addition to killing all O2/O5 strains. This live, attenuated P. aeruginosa strain PAO1 Delta aroA appears to be safe for potential use as an intranasal vaccine and elicits high titers of opsonic antibodies against multiple strains of the P. aeruginosa O2/O5 serogroup.
- Published
- 2002
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14. Transgenic cystic fibrosis mice exhibit reduced early clearance of Pseudomonas aeruginosa from the respiratory tract.
- Author
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Schroeder TH, Reiniger N, Meluleni G, Grout M, Coleman FT, and Pier GB
- Subjects
- Animals, Bacterial Adhesion genetics, Cell Line, Transformed, Cystic Fibrosis pathology, Cystic Fibrosis prevention & control, Cystic Fibrosis Transmembrane Conductance Regulator physiology, Epithelial Cells microbiology, Epithelial Cells ultrastructure, Female, Lung microbiology, Macaca mulatta, Male, Mice, Mice, Inbred BALB C, Mice, Inbred C57BL, Mice, Transgenic, Microscopy, Confocal, Microscopy, Electron, Scanning, Pseudomonas Infections pathology, Pseudomonas Infections prevention & control, Pseudomonas aeruginosa physiology, Pseudomonas aeruginosa ultrastructure, Respiratory Tract Infections pathology, Respiratory Tract Infections prevention & control, Trachea microbiology, Trachea ultrastructure, Cystic Fibrosis genetics, Cystic Fibrosis microbiology, Cystic Fibrosis Transmembrane Conductance Regulator genetics, Pseudomonas Infections genetics, Pseudomonas Infections microbiology, Respiratory Tract Infections genetics, Respiratory Tract Infections microbiology
- Abstract
The cystic fibrosis (CF) transmembrane conductance regulator (CFTR) has been proposed to be an epithelial cell receptor for Pseudomonas aeruginosa involved in bacterial internalization and clearance from the lung. We evaluated the role of CFTR in clearing P. aeruginosa from the respiratory tract using transgenic CF mice that carried either the DeltaF508 Cftr allele or an allele with a Cftr stop codon (S489X). Intranasal application achieved P. aeruginosa lung infection in inbred C57BL/6 DeltaF508 Cftr mice, whereas DeltaF508 Cftr and S489X Cftr outbred mice required tracheal application of the inoculum to establish lung infection. CF mice showed significantly less ingestion of LPS-smooth P. aeruginosa by lung cells and significantly greater bacterial lung burdens 4.5 h postinfection than C57BL/6 wild-type mice. Microscopy of infected mouse and rhesus monkey tracheas clearly demonstrated ingestion of P. aeruginosa by epithelial cells in wild-type animals, mostly around injured areas of the epithelium. Desquamating cells loaded with P. aeruginosa could also be seen in these tissues. No difference was found between CF and wild-type mice challenged with an LPS-rough mucoid isolate of P. aeruginosa lacking the CFTR ligand. Thus, transgenic CF mice exhibit decreased clearance of P. aeruginosa and increased bacterial burdens in the lung, substantiating a key role for CFTR-mediated bacterial ingestion in lung clearance of P. aeruginosa.
- Published
- 2001
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15. Acquisition of expression of the Pseudomonas aeruginosa ExoU cytotoxin leads to increased bacterial virulence in a murine model of acute pneumonia and systemic spread.
- Author
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Allewelt M, Coleman FT, Grout M, Priebe GP, and Pier GB
- Subjects
- Animals, Animals, Newborn, Base Sequence, DNA Primers genetics, Disease Models, Animal, Female, Gene Expression, Humans, Mice, Mice, Inbred BALB C, Pseudomonas aeruginosa isolation & purification, Transformation, Genetic, Virulence genetics, Bacterial Proteins genetics, Cytotoxins genetics, Pneumonia, Bacterial etiology, Pseudomonas Infections etiology, Pseudomonas aeruginosa genetics, Pseudomonas aeruginosa pathogenicity
- Abstract
Pseudomonas aeruginosa is the nosocomial bacterial pathogen most commonly isolated from the respiratory tract. Animal models of this infection are extremely valuable for studies of virulence and immunity. We thus evaluated the utility of a simple model of acute pneumonia for analyzing P. aeruginosa virulence by characterizing the course of bacterial infection in BALB/c mice following application of bacteria to the nares of anesthetized animals. Bacterial aspiration into the lungs was rapid, and 67 to 100% of the inoculum could be recovered within minutes from the lungs, with 0.1 to 1% of the inoculum found intracellularly shortly after infection. At later time points up to 10% of the bacteria were intracellular, as revealed by gentamicin exclusion assays on single-cell suspensions of infected lungs. Expression of exoenzyme U (ExoU) by P. aeruginosa is associated with a cytotoxic effect on epithelial cells in vitro and virulence in animal models. Insertional mutations in the exoU gene confer a noncytotoxic phenotype on mutant strains and decrease virulence for animals. We used the model of acute pneumonia to determine whether introduction of the exoU gene into noncytotoxic strains of P. aeruginosa lacking this gene affected virulence. Seven phenotypically noncytotoxic P. aeruginosa strains were transformed with pUCP19exoUspcU which carries the exoU gene and its associated chaperone. Three of these strains became cytotoxic to cultured epithelial cells in vitro. These strains all secreted ExoU, as confirmed by detection of the ExoU protein with specific antisera. The 50% lethal dose of exoU-expressing strains was significantly lower for all three P. aeruginosa isolates carrying plasmid pUCP19exoUspcU than for the isogenic exoU-negative strains. mRNA specific for ExoU was readily detected in the lungs of animals infected with the transformed P. aeruginosa strains. Introduction of the exoU gene confers a cytotoxic phenotype on some, but not all, otherwise-noncytotoxic P. aeruginosa strains and, for recombinant strains that could express ExoU, there was markedly increased virulence in a murine model of acute pneumonia and systemic spread.
- Published
- 2000
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16. NLRB unit determinations expanding?
- Author
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Coleman FT and Sperling L
- Subjects
- Government Agencies, United States, Collective Bargaining, Health Facilities, Legislation as Topic
- Published
- 1976
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