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  1. Article ; Online: Expanding the Molecular Toolkit for Chlamydia.

    Hybiske, Kevin

    Cell host & microbe

    2015  Volume 18, Issue 1, Page(s) 11–13

    Abstract: Although historically a genetically intractable bacterium, Chlamydia is experiencing a renaissance for molecular genetic manipulation. Two new studies published in Cell Host & Microbe, Mirrashidi et al. (2015) and Kokes et al. (2015), have dramatically ... ...

    Abstract Although historically a genetically intractable bacterium, Chlamydia is experiencing a renaissance for molecular genetic manipulation. Two new studies published in Cell Host & Microbe, Mirrashidi et al. (2015) and Kokes et al. (2015), have dramatically changed the landscape of what is possible for molecular dissection of Chlamydia-host interactions.
    Language English
    Publishing date 2015-07-09
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2278004-X
    ISSN 1934-6069 ; 1931-3128
    ISSN (online) 1934-6069
    ISSN 1931-3128
    DOI 10.1016/j.chom.2015.06.016
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  2. Article: The

    Zuck, Meghan / Hybiske, Kevin

    Microorganisms

    2019  Volume 7, Issue 5

    Abstract: The cellular exit strategies of intracellular pathogens have a direct impact on microbial dissemination, transmission, and engagement of immune responses of the host. ...

    Abstract The cellular exit strategies of intracellular pathogens have a direct impact on microbial dissemination, transmission, and engagement of immune responses of the host.
    Language English
    Publishing date 2019-05-25
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2720891-6
    ISSN 2076-2607
    ISSN 2076-2607
    DOI 10.3390/microorganisms7050149
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  3. Article ; Online: Chlamydia trachomatis seroassays used in epidemiologic research: a narrative review and practical considerations.

    Waters, Mary Bridget / Hybiske, Kevin / Ikeda, Ren / Kaltenboeck, Bernhard / Manhart, Lisa E / Kreisel, Kristen M / Khosropour, Christine M

    The Journal of infectious diseases

    2024  

    Abstract: Chlamydia trachomatis (CT) is a sexually transmitted infection that can lead to adverse reproductive health outcomes. CT prevalence estimates are primarily derived from screening using nucleic acid amplification tests (NAATs). However, screening ... ...

    Abstract Chlamydia trachomatis (CT) is a sexually transmitted infection that can lead to adverse reproductive health outcomes. CT prevalence estimates are primarily derived from screening using nucleic acid amplification tests (NAATs). However, screening guidelines in the United States only include particular subpopulations, and NAATs only detect current infections. In contrast, seroassays identify past CT infections which are important for understanding the public health impacts of CT, including pelvic inflammatory disease and tubal factor infertility. Older seroassays have been plagued by low sensitivity and specificity and have not been validated using a consistent reference measure, making it challenging to compare studies, define the epidemiology of CT and determine the effectiveness of control programs. Newer seroassays have better performance characteristics. This narrative review summarizes the "state of the science" for CT seroassays that have been applied in epidemiologic studies and provides practical considerations for interpreting the literature and employing seroassays in future research.
    Language English
    Publishing date 2024-04-19
    Publishing country United States
    Document type Journal Article
    ZDB-ID 3019-3
    ISSN 1537-6613 ; 0022-1899
    ISSN (online) 1537-6613
    ISSN 0022-1899
    DOI 10.1093/infdis/jiae199
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  4. Article ; Online: Antibodies from chlamydia-infected individuals facilitate phagocytosis via Fc receptors.

    Hybiske, Kevin / Paktinat, Shahrokh / Newman, Katherine / Patton, Dorothy / Khosropour, Christine / Roxby, Alison C / Mugo, Nelly R / Oluoch, Lynda / Ngure, Kenneth / Suchland, Robert / Hladik, Florian / Vojtech, Lucia

    Infection and immunity

    2024  Volume 92, Issue 4, Page(s) e0050323

    Abstract: Non-neutralizing functions of antibodies, including phagocytosis, may play a role ... ...

    Abstract Non-neutralizing functions of antibodies, including phagocytosis, may play a role in
    MeSH term(s) Humans ; Receptors, Fc ; Phagocytosis ; Neutrophils ; Antibodies, Bacterial ; Chlamydia Infections ; Chlamydia trachomatis
    Chemical Substances Receptors, Fc ; Antibodies, Bacterial
    Language English
    Publishing date 2024-03-07
    Publishing country United States
    Document type Journal Article
    ZDB-ID 218698-6
    ISSN 1098-5522 ; 0019-9567
    ISSN (online) 1098-5522
    ISSN 0019-9567
    DOI 10.1128/iai.00503-23
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  5. Article ; Online: The Chlamydia trachomatis Extrusion Exit Mechanism Is Regulated by Host Abscission Proteins

    Meghan Zuck / Kevin Hybiske

    Microorganisms, Vol 7, Iss 5, p

    2019  Volume 149

    Abstract: The cellular exit strategies of intracellular pathogens have a direct impact on microbial dissemination, transmission, and engagement of immune responses of the host. Chlamydia exit their host via a budding mechanism called extrusion, which offers ... ...

    Abstract The cellular exit strategies of intracellular pathogens have a direct impact on microbial dissemination, transmission, and engagement of immune responses of the host. Chlamydia exit their host via a budding mechanism called extrusion, which offers protective benefits to Chlamydia as they navigate their extracellular environment. Many intracellular pathogens co-opt cellular abscission machinery to facilitate cell exit, which is utilized to perform scission of two newly formed daughter cells following mitosis. Similar to viral budding exit strategies, we hypothesize that an abscission-like mechanism is required to physically sever the chlamydial extrusion from the host cell, co-opting the membrane fission activities of the endosomal sorting complex required for transport (ESCRT) family of proteins that are necessary for cellular scission events, including abscission. To test this, C. trachomatis L2-infected HeLa cells were depleted of key abscission machinery proteins charged multivesicle body protein 4b (CHMP4B), ALIX, centrosome protein 55 (CEP55), or vacuolar protein sorting-associated protein 4A (VPS4A), using RNA interference (RNAi). Over 50% reduction in extrusion formation was achieved by depletion of CHMP4B, VPS4A, and ALIX, but no effect on extrusion was observed with CEP55 depletion. These results demonstrate a role for abscission machinery in C. trachomatis extrusion from the host cell, with ALIX, VPS4A and CHMP4B playing key functional roles in optimal extrusion release.
    Keywords Chlamydia trachomatis ; extrusion ; abscission ; Biology (General) ; QH301-705.5
    Subject code 570
    Language English
    Publishing date 2019-05-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: Chlamydia trachomatis Cellular Exit Alters Interactions with Host Dendritic Cells.

    Sherrid, Ashley M / Hybiske, Kevin

    Infection and immunity

    2017  Volume 85, Issue 5

    Abstract: The strategies utilized by pathogens to exit host cells are an area of pathogenesis which has received surprisingly little attention, considering the necessity of this step for infections to propagate. Even less is known about how exit through these ... ...

    Abstract The strategies utilized by pathogens to exit host cells are an area of pathogenesis which has received surprisingly little attention, considering the necessity of this step for infections to propagate. Even less is known about how exit through these pathways affects downstream host-pathogen interactions and the generation of an immune response.
    MeSH term(s) Animals ; Apoptosis ; Cell Survival ; Cells, Cultured ; Chlamydia trachomatis/pathogenicity ; Dendritic Cells/immunology ; Dendritic Cells/microbiology ; Fibroblasts/microbiology ; Host-Pathogen Interactions ; Mice, Inbred C57BL ; Microbial Viability
    Language English
    Publishing date 2017-04-21
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 218698-6
    ISSN 1098-5522 ; 0019-9567
    ISSN (online) 1098-5522
    ISSN 0019-9567
    DOI 10.1128/IAI.00046-17
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  7. Article ; Online: Cellular Exit Strategies of Intracellular Bacteria.

    Hybiske, Kevin / Stephens, Richard

    Microbiology spectrum

    2016  Volume 3, Issue 6

    Abstract: The coevolution of intracellular bacteria with their eukaryotic hosts has presented these pathogens with numerous challenges for their evolutionary progress and survival. Chief among these is the ability to exit from host cells, an event that is ... ...

    Abstract The coevolution of intracellular bacteria with their eukaryotic hosts has presented these pathogens with numerous challenges for their evolutionary progress and survival. Chief among these is the ability to exit from host cells, an event that is fundamentally linked to pathogen dissemination and transmission. Recent years have witnessed a major expansion of research in this area, and this chapter summarizes our current understanding of the spectrum of exit strategies that are exploited by intracellular pathogens. Clear themes regarding the mechanisms of microbial exit have emerged and are most easily conceptualized as (i) lysis of the host cell, (ii) nonlytic exit of free bacteria, and (iii) release of microorganisms into membrane-encased compartments. The adaptation of particular exit strategies is closely linked with additional themes in microbial pathogenesis, including host cell death, manipulation of host signaling pathways, and coincident activation of proinflammatory responses. This chapter will explore the molecular determinants used by intracellular pathogens to promote host cell escape and the infectious advantages each exit pathway may confer, and it will provide an evolutionary framework for the adaptation of these mechanisms.
    MeSH term(s) Animals ; Bacteria/genetics ; Bacteria/growth & development ; Bacterial Infections/genetics ; Bacterial Infections/metabolism ; Bacterial Infections/microbiology ; Bacterial Physiological Phenomena ; Biological Evolution ; Host-Pathogen Interactions ; Humans ; Signal Transduction
    Language English
    Publishing date 2016-06-20
    Publishing country United States
    Document type Journal Article ; Review
    ISSN 2165-0497
    ISSN (online) 2165-0497
    DOI 10.1128/microbiolspec.VMBF-0002-2014
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  8. Article ; Online: A broad-spectrum cloning vector that exists as both an integrated element and a free plasmid in Chlamydia trachomatis.

    Garvin, Lotisha / Vande Voorde, Rebecca / Dickinson, Mary / Carrell, Steven / Hybiske, Kevin / Rockey, Daniel

    PloS one

    2021  Volume 16, Issue 12, Page(s) e0261088

    Abstract: Plasmid transformation of chlamydiae has created new opportunities to investigate host-microbe interactions during chlamydial infections; however, there are still limitations. Plasmid transformation requires a replicon derived from the native Chlamydia ... ...

    Abstract Plasmid transformation of chlamydiae has created new opportunities to investigate host-microbe interactions during chlamydial infections; however, there are still limitations. Plasmid transformation requires a replicon derived from the native Chlamydia plasmid, and these transformations are species-specific. We explored the utility of a broad host-range plasmid, pBBR1MCS-4, to transform chlamydiae, with a goal of simplifying the transformation process. The plasmid was modified to contain chromosomal DNA from C. trachomatis to facilitate homologous recombination. Sequences flanking incA were cloned into the pBBR1MCS-4 vector along with the GFP:CAT cassette from the pSW2-GFP chlamydial shuttle vector. The final plasmid construct, pBVR2, was successfully transformed into C. trachomatis strain L2-434. Chlamydial transformants were analyzed by immunofluorescence microscopy and positive clones were sequentially purified using limiting dilution. PCR and PacBio-based whole genome sequencing were used to determine if the plasmid was maintained within the chromosome or as an episome. PacBio sequencing of the cloned transformants revealed allelic exchange events between the chromosome and plasmid pBVR2 that replaced chromosomal incA with the plasmid GFP:CAT cassette. The data also showed evidence of full integration of the plasmid into the bacterial chromosome. While some plasmids were fully integrated, some were maintained as episomes and could be purified and retransformed into E. coli. Thus, the plasmid can be successfully transformed into chlamydia without a chlamydial origin of replication and can exist in multiple states within a transformed population.
    MeSH term(s) Chlamydia Infections/microbiology ; Chlamydia trachomatis/genetics ; Chlamydia trachomatis/growth & development ; Chromosomes, Bacterial/genetics ; Genetic Vectors ; Green Fluorescent Proteins/genetics ; Green Fluorescent Proteins/metabolism ; Plasmids/genetics ; Transformation, Bacterial
    Chemical Substances Green Fluorescent Proteins (147336-22-9)
    Language English
    Publishing date 2021-12-16
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 2267670-3
    ISSN 1932-6203 ; 1932-6203
    ISSN (online) 1932-6203
    ISSN 1932-6203
    DOI 10.1371/journal.pone.0261088
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  9. Article ; Online: Crystal structure of an inorganic pyrophosphatase from Chlamydia trachomatis D/UW-3/Cx.

    Maddy, Jasmine / Staker, Bart L / Subramanian, Sandhya / Abendroth, Jan / Edwards, Thomas E / Myler, Peter J / Hybiske, Kevin / Asojo, Oluwatoyin A

    Acta crystallographica. Section F, Structural biology communications

    2022  Volume 78, Issue Pt 3, Page(s) 135–142

    Abstract: Chlamydia trachomatis is the leading cause of bacterial sexually transmitted infections globally and is one of the most commonly reported infections in the United States. There is a need to develop new therapeutics due to drug resistance and the failure ... ...

    Abstract Chlamydia trachomatis is the leading cause of bacterial sexually transmitted infections globally and is one of the most commonly reported infections in the United States. There is a need to develop new therapeutics due to drug resistance and the failure of current treatments to clear persistent infections. Structures of potential C. trachomatis rational drug-discovery targets, including C. trachomatis inorganic pyrophosphatase (CtPPase), have been determined by the Seattle Structural Genomics Center for Infectious Disease. Inorganic pyrophosphatase hydrolyzes inorganic pyrophosphate during metabolism. Furthermore, bacterial inorganic pyrophosphatases have shown promise for therapeutic discovery. Here, a 2.2 Å resolution X-ray structure of CtPPase is reported. The crystal structure of CtPPase reveals shared structural features that may facilitate the repurposing of inhibitors identified for bacterial inorganic pyrophosphatases as starting points for new therapeutics for C. trachomatis.
    MeSH term(s) Chlamydia trachomatis/metabolism ; Crystallography, X-Ray ; Inorganic Pyrophosphatase/metabolism ; United States
    Chemical Substances Inorganic Pyrophosphatase (EC 3.6.1.1)
    Language English
    Publishing date 2022-02-28
    Publishing country United States
    Document type Journal Article
    ISSN 2053-230X
    ISSN (online) 2053-230X
    DOI 10.1107/S2053230X22002138
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  10. Article ; Online: Genomic Analysis of MSM Rectal Chlamydia trachomatis Isolates Identifies Predicted Tissue-Tropic Lineages Generated by Intraspecies Lateral Gene Transfer-Mediated Evolution.

    Suchland, Robert J / Carrell, Steven J / Ramsey, Stephen A / Hybiske, Kevin / Debrine, Abigail M / Sanchez, Jorge / Celum, Connie / Rockey, Daniel D

    Infection and immunity

    2022  Volume 90, Issue 11, Page(s) e0026522

    Abstract: Chlamydia trachomatis is an obligate intracellular bacterium that causes serious diseases in humans. Rectal infection and disease caused by this pathogen are important yet understudied aspects of C. trachomatis natural history. The University of ... ...

    Abstract Chlamydia trachomatis is an obligate intracellular bacterium that causes serious diseases in humans. Rectal infection and disease caused by this pathogen are important yet understudied aspects of C. trachomatis natural history. The University of Washington Chlamydia Repository has a large collection of male-rectal-sourced strains (MSM rectal strains) isolated in Seattle, USA and Lima, Peru. Initial characterization of strains collected over 30 years in both Seattle and Lima led to an association of serovars G and J with male rectal infections. Serovar D, E, and F strains were also collected from MSM patients. Genome sequence analysis of a subset of MSM rectal strains identified a clade of serovar G and J strains that had high overall genomic identity. A genome-wide association study was then used to identify genomic loci that were correlated with tissue tropism in a collection of serovar-matched male rectal and female cervical strains. The polymorphic membrane protein PmpE had the strongest correlation, and amino acid sequence alignments identified a set of PmpE variable regions (VRs) that were correlated with host or tissue tropism. Examination of the positions of VRs by the protein structure-predicting Alphafold2 algorithm demonstrated that the VRs were often present in predicted surface-exposed loops in both PmpE and PmpH protein structure. Collectively, these studies identify possible tropism-predictive loci for MSM rectal C. trachomatis infections and identify predicted surface-exposed variable regions of Pmp proteins that may function in MSM rectal versus cervical tropism differences.
    MeSH term(s) Humans ; Male ; Chlamydia Infections/microbiology ; Chlamydia trachomatis/genetics ; Gene Transfer, Horizontal ; Genome-Wide Association Study ; Genomics ; Homosexuality, Male
    Language English
    Publishing date 2022-10-10
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Extramural
    ZDB-ID 218698-6
    ISSN 1098-5522 ; 0019-9567
    ISSN (online) 1098-5522
    ISSN 0019-9567
    DOI 10.1128/iai.00265-22
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