Veillonellaceae family members uniquely alter the cervical metabolic microenvironment in a human three-dimensional epithelial model

AbstractBacterial vaginosis (BV) is a gynecologic disorder characterized by a shift in cervicovaginal microbiota from Lactobacillus spp. dominance to a polymicrobial biofilm composed of diverse anaerobes. We utilized a well-characterized human three-dimensional cervical epithelial cell model in conjunction with untargeted metabolomics and immunoproteomics analyses to determine the immunometabolic contribution of three members of the Veillonellaceae family: Veillonella atypica, Veillonella montpellierensis and Megasphaera micronuciformis at this site. We found that Veillonella spp. infections induced significant elevation of polyamines. M. micronuciformis infections significantly increased soluble inflammatory mediators, induced moderate levels of cell cytotoxicity, and accumulation of cell membrane lipids relative to Veillonella spp. Notably, both V. atypica and V. montpellierensis infections resulted in consumption of lactate, a key metabolite linked to gynecologic and reproductive health. Collectively our approach and data provide unique insights into the specific contributions of Veillonellaceae members to the pathogenesis of BV and women’s health.

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Pleiotropic effects of FGFR1 on cell proliferation, survival, and migration in a 3D mammary epithelial cell model

The Journal of Cell Biology ◽

10.1083/jcb.200505098 ◽

2005 ◽

Vol 171 (4) ◽

pp. 663-673 ◽

Cited By ~ 89

Author(s):

Wa Xian ◽

Kathryn L. Schwertfeger ◽

Tracy Vargo-Gogola ◽

Jeffrey M. Rosen

Keyword(s):

Cell Proliferation ◽

Epithelial Cell ◽

Mammary Epithelial Cell ◽

Molecular Mechanisms ◽

Cell Model ◽

Smooth Muscle Actin ◽

Three Dimensional ◽

Mammary Epithelial ◽

Matrix Metalloproteinase 3

Members of the fibroblast growth factor (FGF) family and the FGF receptors (FGFRs) have been implicated in mediating various aspects of mammary gland development and transformation. To elucidate the molecular mechanisms of FGFR1 action in a context that mimics polarized epithelial cells, we have developed an in vitro three-dimensional HC11 mouse mammary epithelial cell culture model expressing a drug-inducible FGFR1 (iFGFR1). Using this conditional model, iFGFR1 activation in these growth-arrested and polarized mammary acini initially led to reinitiation of cell proliferation, increased survival of luminal cells, and loss of cell polarity, resulting in the disruption of acinar structures characterized by the absence of an empty lumen. iFGFR1 activation also resulted in a gain of invasive properties and the induction of matrix metalloproteinase 3 (MMP-3), causing the cleavage of E-cadherin and increased expression of smooth muscle actin and vimentin. The addition of a pan MMP inhibitor abolished these phenotypes but did not prevent the effects of iFGFR1 on cell proliferation or survival.

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Analysis of Host Responses to Neisseria gonorrhoeae Using a Human Three-Dimensional Endometrial Epithelial Cell Model

Neisseria gonorrhoeae - Methods in Molecular Biology ◽

10.1007/978-1-4939-9496-0_20 ◽

2019 ◽

pp. 347-361 ◽

Cited By ~ 1

Author(s):

Paweł Łaniewski ◽

Melissa M. Herbst-Kralovetz

Keyword(s):

Epithelial Cell ◽

Neisseria Gonorrhoeae ◽

Cell Model ◽

Three Dimensional ◽

Host Responses ◽

Endometrial Epithelial Cell

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Interleukin-36γ Is Elevated in Cervicovaginal Epithelial Cells in Women With Bacterial Vaginosis and In Vitro After Infection With Microbes Associated With Bacterial Vaginosis

The Journal of Infectious Diseases ◽

10.1093/infdis/jiz514 ◽

2019 ◽

Vol 221 (6) ◽

pp. 983-988 ◽

Cited By ~ 4

Author(s):

Jameson K Gardner ◽

Paweł Łaniewski ◽

Anna Knight ◽

Lisa B Haddad ◽

Alison Swaims-Kohlmeier ◽

...

Keyword(s):

Epithelial Cell ◽

Epithelial Cells ◽

Bacterial Vaginosis ◽

Host Response ◽

Cell Model ◽

Bacterial Species ◽

Clinical Samples ◽

Sexually Transmitted ◽

3 Dimensional

Abstract In recent studies, the interleukin (IL)-36 cytokines were shown to be elevated in women with non-Lactobacillus-dominated vaginal microbiomes. In this study, we evaluated IL36G expression in clinical samples from women with and without bacterial vaginosis (BV) and a human 3-dimensional cervical epithelial cell model. IL36G expression was significantly elevated in cervicovaginal epithelial cells isolated from BV-positive women and corresponded with increased neutrophil counts relative to BV-negative women. In addition, specific BV-associated bacterial species as well as a polymicrobial cocktail significantly induced IL36G expression in vitro. These findings suggest that IL-36γ may exhibit an important function in the host response to BV and other sexually transmitted infections.

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Bacterial vaginosis and health-associated bacteria modulate the immunometabolic landscape in 3D model of human cervix

npj Biofilms and Microbiomes ◽

10.1038/s41522-021-00259-8 ◽

2021 ◽

Vol 7 (1) ◽

Author(s):

Paweł Łaniewski ◽

Melissa M. Herbst-Kralovetz

Keyword(s):

Epithelial Cell ◽

Bacterial Vaginosis ◽

Pathogenic Bacteria ◽

Cell Model ◽

Female Reproductive Tract ◽

Gardnerella Vaginalis ◽

Obstetric Outcomes ◽

Phenyllactic Acid ◽

Increased Risk ◽

Human Cervix

AbstractBacterial vaginosis (BV) is an enigmatic polymicrobial condition characterized by a depletion of health-associated Lactobacillus and an overgrowth of anaerobes. Importantly, BV is linked to adverse gynecologic and obstetric outcomes: an increased risk of sexually transmitted infections, preterm birth, and cancer. We hypothesized that members of the cervicovaginal microbiota distinctly contribute to immunometabolic changes in the human cervix, leading to these sequelae. Our 3D epithelial cell model that recapitulates the human cervical epithelium was infected with clinical isolates of cervicovaginal bacteria, alone or as a polymicrobial community. We used Lactobacillus crispatus as a representative health-associated commensal and four common BV-associated species: Gardnerella vaginalis, Prevotella bivia, Atopobium vaginae, and Sneathia amnii. The immunometabolic profiles of these microenvironments were analyzed using multiplex immunoassays and untargeted global metabolomics. A. vaginae and S. amnii exhibited the highest proinflammatory potential through induction of cytokines, iNOS, and oxidative stress-associated compounds. G. vaginalis, P. bivia, and S. amnii distinctly altered physicochemical barrier-related proteins and metabolites (mucins, sialic acid, polyamines), whereas L. crispatus produced an antimicrobial compound, phenyllactic acid. Alterations to the immunometabolic landscape correlate with symptoms and hallmarks of BV and connected BV with adverse women’s health outcomes. Overall, this study demonstrated that 3D cervical epithelial cell colonized with cervicovaginal microbiota faithfully reproduce the immunometabolic microenvironment previously observed in clinical studies and can successfully be used as a robust tool to evaluate host responses to commensal and pathogenic bacteria in the female reproductive tract.

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Immunometabolic Analysis of Mobiluncus mulieris and Eggerthella sp. Reveals Novel Insights Into Their Pathogenic Contributions to the Hallmarks of Bacterial Vaginosis

Frontiers in Cellular and Infection Microbiology ◽

10.3389/fcimb.2021.759697 ◽

2021 ◽

Vol 11 ◽

Author(s):

Ross McKenzie ◽

Jason D. Maarsingh ◽

Paweł Łaniewski ◽

Melissa M. Herbst-Kralovetz

Keyword(s):

Oxidative Stress ◽

Energy Metabolism ◽

Bacterial Vaginosis ◽

Biogenic Amines ◽

Cell Model ◽

Three Dimensional ◽

Reproductive Age ◽

Pathogenic Microorganisms ◽

Epithelial Barrier Function ◽

And Oxidative Stress

The cervicovaginal microbiome plays an important role in protecting women from dysbiosis and infection caused by pathogenic microorganisms. In healthy reproductive-age women the cervicovaginal microbiome is predominantly colonized by protective Lactobacillus spp. The loss of these protective bacteria leads to colonization of the cervicovaginal microenvironment by pathogenic microorganisms resulting in dysbiosis and bacterial vaginosis (BV). Mobiluncus mulieris and Eggerthella sp. are two of the many anaerobes that can contribute to BV, a condition associated with multiple adverse obstetric and gynecological outcomes. M. mulieris has been linked to high Nugent scores (relating to BV morphotypes) and preterm birth (PTB), whilst some bacterial members of the Eggerthellaceae family are highly prevalent in BV, and identified in ~85-95% of cases. The functional impact of M. mulieris and Eggerthella sp. in BV is still poorly understood. To determine the individual immunometabolic contributions of Eggerthella sp. and M. mulieris within the cervicovaginal microenvironment, we utilized our well-characterized human three-dimensional (3-D) cervical epithelial cell model in combination with multiplex immunoassays and global untargeted metabolomics approaches to identify key immune mediators and metabolites related to M. mulieris and Eggerthella sp. infections. We found that infection with M. mulieris significantly elevated multiple proinflammatory markers (IL-6, IL-8, TNF-α and MCP-1) and altered metabolites related to energy metabolism (nicotinamide and succinate) and oxidative stress (cysteinylglycine, cysteinylglycine disulfide and 2-hydroxygluatrate). Eggerthella sp. infection significantly elevated multiple sphingolipids and glycerolipids related to epithelial barrier function, and biogenic amines (putrescine and cadaverine) associated with elevated vaginal pH, vaginal amine odor and vaginal discharge. Our study elucidated that M. mulieris elevated multiple proinflammatory markers relating to PTB and STI acquisition, as well as altered energy metabolism and oxidative stress, whilst Eggerthella sp. upregulated multiple biogenic amines associated with the clinical diagnostic criteria of BV. Future studies are needed to evaluate how these bacteria interact with other BV-associated bacteria within the cervicovaginal microenvironment.

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Study on erythrocytes by SEM photogrammetry (SEMP) technique

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100161187 ◽

1990 ◽

Vol 48 (3) ◽

pp. 724-725

Author(s):

Tong Wensheng ◽

Lu Lianhuang ◽

Zhang Zhijun

Keyword(s):

Quantitative Analysis ◽

Cell Membrane ◽

Clinical Diagnosis ◽

Human Body ◽

Blood Cells ◽

Three Dimensional ◽

Normal Person ◽

Blood Disease ◽

Computation Technique ◽

Important Basis

This is a combined study of two diffirent branches, photogrammetry and morphology of blood cells. The three dimensional quantitative analysis of erythrocytes using SEMP technique, electron computation technique and photogrammetry theory has made it possible to push the study of mophology of blood cells from LM, TEM, SEM to a higher stage, that of SEM P. A new path has been broken for deeply study of morphology of blood cells.In medical view, the abnormality of the quality and quantity of erythrocytes is one of the important changes of blood disease. It shows the abnormal blood—making function of the human body. Therefore, the study of the change of shape on erythrocytes is the indispensable and important basis of reference in the clinical diagnosis and research of blood disease.The erythrocytes of one normal person, three PNH Patients and one AA patient were used in this experiment. This research determines the following items: Height;Length of two axes (long and short), ratio; Crevice in depth and width of cell membrane; Circumference of erythrocytes; Isoline map of erythrocytes; Section map of erythrocytes.

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