@article {848,
	title = {In vitro assessment of halobacterial gas vesicles as a Chlamydia vaccine display and delivery system.},
	journal = {Vaccine},
	volume = {30},
	year = {2012},
	month = {2012 Sep 7},
	pages = {5942-8},
	abstract = {Chlamydia trachomatis is the leading cause of bacterial sexually transmitted disease worldwide and while antibiotic treatment is effective in eliminating the pathogen, up to 70\% of all infections are asymptomatic. Despite sustained efforts over the past 2 decades, an effective chlamydial vaccine remains elusive, due in large part to the lack of an effective delivery system. We explored the use of gas vesicles derived from Halobacterium salinarium as a potential display and delivery vehicle for chlamydial antigens of vaccine interest. Various size gene fragments coding for the major outer membrane protein (MOMP), outer membrane complex B (OmcB) and polymorphic outer membrane protein D (PompD) were integrated into and expressed as part of the gas vesicle protein C (gvpC) on the surface of these stable structures. The presence of the recombinant proteins was confirmed by Western blots probed using anti-gvpC and anti-Chlamydia antibodies as well as sera from Chlamydia-positive patients. Tissue culture evaluation revealed stability and a time-dependent degradation of recombinant gas vesicles (r-Gv) in human and animal cell lines. In vitro assessment using human foreskin fibroblasts (HFF) confirmed Toll-like receptor (TLR) 4 and 5 engagement by wild type and r-Gv, leading to MyD88 activation, TNF-α, IL-6 and IL-12 production. The data suggest that r-GV could be an effective, naturally adjuvanting, time-release antigen delivery system for immunologically relevant Chlamydia vaccine antigens which are readily recognized by human immune sera.},
	issn = {1873-2518},
	doi = {10.1016/j.vaccine.2012.07.038},
	author = {Childs, Tawanna S and Webley, Wilmore C}
}
@article {849,
	title = {The prevalence and identity of Chlamydia-specific IgE in children with asthma and other chronic respiratory symptoms.},
	journal = {Respir Res},
	volume = {13},
	year = {2012},
	month = {2012 Apr 18},
	pages = {32},
	abstract = {ABSTRACT: BACKGROUND: Recent studies have confirmed the presence of viable Chlamydia in the bronchoalveolar lavage (BAL) fluid of pediatric patients with airway hyperresponsiveness. While specific IgG and IgM responses to C. pneumoniae are well described, the response and potential contribution of Ag-specific IgE are not known. The current study sought to determine if infection with Chlamydia triggers the production of pathogen-specific IgE in children with chronic respiratory diseases which might contribute to inflammation and pathology. METHODS: We obtained BAL fluid and serum from pediatric respiratory disease patients who were generally unresponsive to corticosteroid treatment as well as sera from age-matched control patients who saw their doctor for wellness checkups. Chlamydia-specific IgE was isolated from BAL and serum samples and their specificity determined by Western blot techniques. The presence of Chlamydia was confirmed by species-specific PCR and BAL culture assays. RESULTS: Chlamydial DNA was detected in the BAL fluid of 134/197 (68\%) patients. Total IgE increased with age until 15 years old and then decreased. Chlamydia-specific IgE was detected in the serum and/or BAL of 107/197 (54\%) patients suffering from chronic respiratory disease, but in none of the 35 healthy control sera (p < 0.0001). Of the 74 BAL culture-positive patients, 68 (91.9\%, p = 0.0001) tested positive for Chlamydia-specific IgE. Asthmatic patients had significantly higher IgE levels compared to non-asthmatics (p = 0.0001). Patients who were positive for Chlamydia DNA or culture had significantly higher levels of serum IgE compared to negative patients (p = 0.0071 and p = 0.0001 respectively). Only 6 chlamydial antigens induced Chlamydia-specific IgE and patients with C. pneumoniae-specific IgE had significantly greater levels of total IgE compared to C. pneumoniae-specific IgE negative ones (p = 0.0001). CONCLUSIONS: IgE antibodies play a central role in allergic inflammation; therefore production of Chlamydia-specific IgE may prove significant in the exacerbation of chronic, allergic airway diseases, thus highlighting a direct role for Chlamydia in asthma pathogenesis.},
	issn = {1465-993X},
	doi = {10.1186/1465-9921-13-32},
	author = {Patel, Katir K and Anderson, Erica A and Salva, Paul S and Webley, Wilmore C}
}
@article {851,
	title = {Colonization of paediatric lower respiratory tract with genital Mycoplasma species.},
	journal = {Respirology},
	volume = {16},
	year = {2011},
	month = {2011 Oct},
	pages = {1081-7},
	abstract = {BACKGROUND AND OBJECTIVE: Recently, much attention has been given to the possible role played by pathogens that colonize neonatal or paediatric airway and their potential involvement in chronic respiratory disease. The goal of the current study was to evaluate the prevalence of Mycoplasma organisms in the BAL fluid of paediatric patients suffering from a variety of chronic respiratory diseases to determine if there was any clear disease association with bacterial presence.

METHODS: We examined 319 paediatric BAL samples for the presence of M.genitalium, M.hominis, U.urealyticum, U.parvum and M.pneumoniae DNA with species-specific PCR.

RESULTS: Mycoplasma DNA was found in 32.6\% (104/319) of patient samples; 10\% (32/319) for M.pneumoniae, 8.8\% for U.parvum, 2.8\% for U.urealyticum; 4.7\% for M.hominis and 9.1\% for M.genitalium. There were no significant clinical and laboratory differences except serum IgE in asthmatics according to Mycoplasma colonization or not. Elevated levels of IgE were found more often in Mycoplasma DNA-negative patients than patients with bacterial DNA, 85/109 versus 24/109 respectively (P<0.0001). There was no difference in the frequency of Mycoplasmas between the asthmatics and the non-asthmatics; 30.6\% (69/225) versus 37.2\% (35/94) for Mycoplasma 16S DNA, and 8\% versus 14.9\% for M.pneumoniae, respectively.

CONCLUSIONS: Our data indicate that in addition to M.pneumoniae, urogenital Mycoplasma species may colonize the airway of patients with chronic respiratory diseases. There was, however, no association between chronic asthma diagnosis and Mycoplasma colonization in this study.},
	keywords = {Adolescent, Asthma, Child, Child, Preschool, Cohort Studies, DNA, Bacterial, Female, Humans, Infant, Infant, Newborn, Male, Mycoplasma, Mycoplasma genitalium, Mycoplasma hominis, Mycoplasma Infections, Mycoplasma pneumoniae, Respiratory System, Species Specificity, Ureaplasma, Ureaplasma Infections, Ureaplasma urealyticum, Young Adult},
	issn = {1440-1843},
	doi = {10.1111/j.1440-1843.2011.02016.x},
	author = {Patel, Katir K and Salva, Paul S and Webley, Wilmore C}
}
@article {852,
	title = {Infectious Chlamydia pneumoniae is associated with elevated interleukin-8 and airway neutrophilia in children with refractory asthma.},
	journal = {Pediatr Infect Dis J},
	volume = {29},
	year = {2010},
	month = {2010 Dec},
	pages = {1093-8},
	abstract = {BACKGROUND: Neutrophilic asthma is thought to be less responsive than eosinophilic asthma to anti-inflammatory therapies including corticosteroids. Chlamydia pneumoniae has been implicated in asthma, possibly by induction of interleukin (IL-8). We hypothesized that IL-8 is increased in the bronchoalveolar lavage (BAL) fluid from children with asthma and C. pneumoniae.

METHODS: BAL fluid was analyzed for C. pneumoniae and IL-8 using polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay from 2 asthma patient populations in the Bronx, NY and Massachusetts with an average age of 8 and 8.7 years old, respectively. For comparison, samples were also analyzed for C. trachomatis and Mycoplasma 16s DNA.

RESULTS: Of 18 Bronx samples analyzed, 6 (33\%) were PCR-positive for C. pneumoniae, 10 (56\%) for C. trachomatis, and 8 (44\%) for Mycoplasma 16s DNA. IL-8 from C. pneumoniae-positive samples was 3.3-fold higher compared with negative samples (P = 0.003). There was no difference between patients tested for C. trachomatis or Mycoplasma. Of 84 Massachusetts samples analyzed, 42 (50\%) were PCR-positive for C. pneumoniae, 42 (50\%) for C. trachomatis, and 13 (16\%) for Mycoplasma. IL-8 concentration from C. pneumoniae-positive samples was 10.49-fold higher compared with negative samples (P = 0.0001). As in the Bronx cohort, there were no differences between patients tested for C. trachomatis or Mycoplasma. Lastly, BAL neutrophilia predicted the presence of C. pneumoniae but not Mycoplasma or C. trachomatis.

CONCLUSIONS: Children with asthma who were PCR-positive for C. pneumoniae demonstrated elevated concentrations of IL-8 and neutrophils in BAL fluid compared with similar patients who were positive for C. trachomatis or Mycoplasma organisms, but PCR-negative for C. pneumoniae. Undiagnosed C. pneumoniae infection in children may therefore contribute to poorly controlled asthma via induction of IL-8.},
	keywords = {Asthma, Bronchoalveolar Lavage Fluid, Child, Chlamydia trachomatis, Chlamydophila Infections, Chlamydophila pneumoniae, DNA, Bacterial, DNA, Ribosomal, Enzyme-Linked Immunosorbent Assay, Female, Humans, Interleukin-8, Male, Massachusetts, Mycoplasma, Neutrophils, New York City, Polymerase Chain Reaction, Respiratory System, RNA, Ribosomal, 16S},
	issn = {1532-0987},
	author = {Patel, Katir K and Vicencio, Alfin G and Du, Zhongfang and Tsirilakis, Kalliope and Salva, Paul S and Webley, Wilmore C}
}
@article {854,
	title = {Notice of duplicate publication.},
	journal = {Am J Respir Crit Care Med},
	volume = {175},
	year = {2007},
	month = {2007 Jan 1},
	pages = {94},
	keywords = {Duplicate Publication as Topic},
	issn = {1073-449X},
	author = {Webley, Wilmore C and Salva, Paul S and Andrzejewski, Chester and Cirino, Frances and West, Corrie A and Tilahun, Yaphet and Stuart, Elizabeth S}
}
@article {855,
	title = {Detection of Chlamydia in the peripheral blood cells of normal donors using in vitro culture, immunofluorescence microscopy and flow cytometry techniques.},
	journal = {BMC Infect Dis},
	volume = {6},
	year = {2006},
	month = {2006},
	pages = {23},
	abstract = {BACKGROUND: Chlamydia trachomatis (Ct) and Chlamydia pneumoniae (Cp) are medically significant infectious agents associated with various chronic human pathologies. Nevertheless, specific roles in disease progression or initiation are incompletely defined. Both pathogens infect established cell lines in vitro and polymerase chain reaction (PCR) has detected Chlamydia DNA in various clinical specimens as well as in normal donor peripheral blood monocytes (PBMC). However, Chlamydia infection of other blood cell types, quantification of Chlamydia infected cells in peripheral blood and transmission of this infection in vitro have not been examined.

METHODS: Cp specific titers were assessed for sera from 459 normal human donor blood (NBD) samples. Isolated white blood cells (WBC) were assayed by in vitro culture to evaluate infection transmission of blood cell borne chlamydiae. Smears of fresh blood samples (FB) were dual immunostained for microscopic identification of Chlamydia-infected cell types and aliquots also assessed using Flow Cytometry (FC).

RESULTS: ELISA demonstrated that 219 (47.7\%) of the NBD samples exhibit elevated anti-Cp antibody titers. Imunofluorescence microscopy of smears demonstrated 113 (24.6\%) of samples contained intracellular Chlamydia and monoclonals to specific CD markers showed that in vivo infection of neutrophil and eosinophil/basophil cells as well as monocytes occurs. In vitro culture established WBCs of 114 (24.8\%) of the NBD samples harbored infectious chlamydiae, clinically a potentially source of transmission, FC demonstrated both Chlamydia infected and uninfected cells can be readily identified and quantified.

CONCLUSION: NBD can harbor infected neutrophils, eosinophil/basophils and monocytes. The chlamydiae are infectious in vitro, and both total, and cell type specific Chlamydia carriage is quantifiable by FC.},
	keywords = {Adult, Blood Donors, Chlamydophila pneumoniae, Female, Flow Cytometry, Humans, Leukocytes, Male, Microscopy, Fluorescence},
	issn = {1471-2334},
	doi = {10.1186/1471-2334-6-23},
	author = {Cirino, Frances and Webley, Wilmore C and West, Corrie and Croteau, Nancy L and Andrzejewski, Chester and Stuart, Elizabeth S}
}
@article {856,
	title = {The bronchial lavage of pediatric patients with asthma contains infectious Chlamydia.},
	journal = {Am J Respir Crit Care Med},
	volume = {171},
	year = {2005},
	month = {2005 May 15},
	pages = {1083-8},
	abstract = {There has been a worldwide increase in the incidence of asthma, and the disease has greatly impacted the public health care system. Chlamydia pneumoniae has been reported as a possible contributing factor in asthma. The organism has been detected by polymerase chain reaction (PCR) in bronchial tissue, but there has been no direct evidence of viability. To determine the frequency of viable Chlamydia in children, blood and bronchoalveolar lavage were collected from 70 pediatric patients undergoing flexible fiberoptic bronchoscopy. Forty-two of these patients had asthma, whereas the remaining patients had various respiratory disorders. Fifty-four percent (38) of the bronchoalveolar lavage samples were PCR-positive for Chlamydia, and 31\% (22) of the PCR-positive samples were positive when cultured on macrophages. Twenty-eight samples (40\%) and 14 samples (20\%) of the PCR- and culture-positive samples, respectively, were from patients with asthma. Culture of the blood samples revealed that 24 (34.3\%) of 70 were positive for Chlamydia compared with 8 (11\%) of 70 matched nonrespiratory control subjects (p < 0.01); 17 (24\%) of the positive blood cultures from the respiratory group were from patients with asthma. Elevation of total IgE was strongly associated with lavage culture positivity for Chlamydia. We therefore conclude that viable Chlamydia pneumoniae organisms are frequently present in the lung lavage fluid from this cohort of predominantly asthmatic pediatric patients.},
	keywords = {Adolescent, Adult, Asthma, Bronchoalveolar Lavage Fluid, Child, Child, Preschool, Chlamydia, Chlamydia Infections, Cohort Studies, Female, Humans, Immunoglobulin E, Infant, Infant, Newborn, Male, Polymerase Chain Reaction, Tissue Culture Techniques},
	issn = {1073-449X},
	doi = {10.1164/rccm.200407-917OC},
	author = {Webley, Wilmore C and Salva, Paul S and Andrzejewski, Chester and Cirino, Frances and West, Corrie A and Tilahun, Yaphet and Stuart, Elizabeth S}
}
@article {716,
	title = {Caveolin-2 associates with intracellular chlamydial inclusions independently of caveolin-1.},
	journal = {BMC Infect Dis},
	volume = {4},
	year = {2004},
	month = {2004 Jul 22},
	pages = {23},
	abstract = {BACKGROUND: Lipid raft domains form in plasma membranes of eukaryotic cells by the tight packing of glycosphingolipids and cholesterol. Caveolae are invaginated structures that form in lipid raft domains when the protein caveolin-1 is expressed. The Chlamydiaceae are obligate intracellular bacterial pathogens that replicate entirely within inclusions that develop from the phagocytic vacuoles in which they enter. We recently found that host cell caveolin-1 is associated with the intracellular vacuoles and inclusions of some chlamydial strains and species, and that entry of those strains depends on intact lipid raft domains. Caveolin-2 is another member of the caveolin family of proteins that is present in caveolae, but of unknown function.

METHODS: We utilized a caveolin-1 negative/caveolin-2 positive FRT cell line and laser confocal immunofluorescence techniques to visualize the colocalization of caveolin-2 with the chlamydial inclusions.

RESULTS: We show here that in infected HeLa cells, caveolin-2, as well as caveolin-1, colocalizes with inclusions of C. pneumoniae (Cp), C. caviae (GPIC), and C. trachomatis serovars E, F and K. In addition, caveolin-2 also associates with C. trachomatis serovars A, B and C, although caveolin-1 did not colocalize with these organisms. Moreover, caveolin-2 appears to be specifically, or indirectly, associated with the pathogens at the inclusion membranes. Using caveolin-1 deficient FRT cells, we show that although caveolin-2 normally is not transported out of the Golgi in the absence of caveolin-1, it nevertheless colocalizes with chlamydial inclusions in these cells. However, our results also show that caveolin-2 did not colocalize with UV-irradiated Chlamydia in FRT cells, suggesting that in these caveolin-1 negative cells, pathogen viability and very likely pathogen gene expression are necessary for the acquisition of caveolin-2 from the Golgi.

CONCLUSION: Caveolin-2 associates with the chlamydial inclusion independently of caveolin-1. The function of caveolin-2, either in the uninfected cell or in the chlamydial developmental cycle, remains to be elucidated. Nevertheless, this second caveolin protein can now be added to the small number of host proteins that are associated with the inclusions of this obligate intracellular pathogen.},
	keywords = {Animals, Blotting, Western, Caveolin 1, Caveolin 2, Caveolins, Cells, Cultured, Chlamydia, Fluorescent Antibody Technique, Golgi Apparatus, Guinea Pigs, HeLa Cells, Humans, Mice, Microscopy, Confocal, Rats, Vacuoles, Virulence},
	issn = {1471-2334},
	doi = {10.1186/1471-2334-4-23},
	author = {Webley, Wilmore C and Norkin, Leonard C and Stuart, Elizabeth S}
}
@article {857,
	title = {Cell surface display of the chlamydial glycolipid exoantigen (GLXA) demonstrated by antibody-dependent complement-mediated cytotoxicity.},
	journal = {Curr Microbiol},
	volume = {49},
	year = {2004},
	month = {2004 Jul},
	pages = {13-21},
	abstract = {The chlamydial species are Gram-negative bacterial pathogens critical to human health. Their developmental cycle is associated with the formation and release of the broadly conserved glycolipid exoantigen (GLXA), which has been implicated in the chlamydial elementary body-host cell interaction. This study examines the potential surface display of this glycolipid by chlamydiae-infected cells and the ability of the GLXA they secrete to associate with the plasma membranes of uninfected cells, a prerequisite for exerting influence on them. The sequential incubation of anti-GLXA antibody and complement with Chlamydia trachomatis serovar K or C. pneumoniae AR-39-infected HeLa 229 or macrophage cells resulted in significant cellular cytotoxicity, which preceded the formation of mature elementary bodies. For uninfected cells, co-incubation of GLXA, purified from supernatants of either C. trachomatis or C. pneumoniae-infected HeLa 229 cells, followed by the successive addition of mouse anti-GLXA antibody and complement, yielded similar levels of cellular cytotoxicity. Thus, GLXA indeed is displayed on the surface of infected cells and, therefore, if antibody of appropriate specificity were present, this GLXA could serve to target these infected cells for elimination. Furthermore, released GLXA can associate with uninfected cells and therefore would be positioned to influence their behavior, especially in the context of infection.},
	keywords = {Antibody Specificity, Chlamydia, Complement System Proteins, Glycolipids, HeLa Cells, Humans, Polysaccharides, Bacterial},
	issn = {0343-8651},
	doi = {10.1007/s00284-003-4181-7},
	author = {Webley, Wilmore C and Vora, Gary J and Stuart, Elizabeth S}
}
@article {715,
	title = {Lipid rafts, caveolae, caveolin-1, and entry by Chlamydiae into host cells.},
	journal = {Exp Cell Res},
	volume = {287},
	year = {2003},
	month = {2003 Jul 1},
	pages = {67-78},
	abstract = {Obligate intracellular bacterial pathogens of the genus Chlamydia are reported to enter host cells by both clathrin-dependent and clathrin-independent processes. C. trachomatis serovar K recently was shown to enter cells via caveolae-like lipid raft domains. We asked here how widespread raft-mediated entry might be among the Chlamydia. We show that C. pneumoniae, an important cause of respiratory infections in humans that additionally is associated with cardiovascular disease, and C. psittaci, an important pathogen in domestic mammals and birds that also infects humans, each enter host cells via cholesterol-rich lipid raft microdomains. Further, we show that C. trachomatis serovars E and F also use these domains to enter host cells. The involvement of these membrane domains in the entry of these organisms was indicated by the sensitivity of their entry to the raft-disrupting agents Nystatin and filipin, and by their intracellular association with caveolin-1, a 22-kDa protein associated with the formation of caveolae in rafts. In contrast, caveolin-marked lipid raft domains do not mediate entry of C. trachomatis serovars A, 36B, and C, nor of LGV serovar L2 and MoPn. Finally, we show that entry of each of these chlamydial strains is independent of cellular expression of caveolin-1. Thus, entry via the Nystatin and filipin-sensitive pathway is dependent on lipid rafts containing cholesterol, rather than invaginated caveolae per se.},
	keywords = {Anti-Bacterial Agents, Caveolae, Caveolin 1, Caveolins, Chlamydia, Chlamydia Infections, Cholesterol, Dose-Response Relationship, Drug, Eukaryotic Cells, Filipin, HeLa Cells, Host-Parasite Interactions, Humans, Membrane Microdomains, Nystatin, Species Specificity},
	issn = {0014-4827},
	author = {Stuart, Elizabeth S and Webley, Wilmore C and Norkin, Leonard C}
}
