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Cervical mucins carry α(1,2)fucosylated glycans that partly protect from experimental vaginal candidiasis

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Abstract

Cervical mucins are glycosylated proteins that form a protective cervical mucus. To understand the role of mucin glycans in Candida albicans infection, oligosaccharides from mouse cervical mucins were analyzed by liquid chromatography-mass spectrometry. Cervical mucins carry multiple α(1-2)fucosylated glycans, but α(1,2)fucosyltransferase Fut2-null mice are devoid of these epitopes. Epithelial cells in vaginal lavages from Fut2-null mice lacked Ulex europaeus agglutinin-1 (UEA-I) staining for α(1-2)fucosylated glycans. Hysterectomy to remove cervical mucus eliminated UEA-I and acid mucin staining in vaginal epithelial cells from wild type mice indicating the cervix as the source of UEA-I positive epithelial cells. To assess binding of α(1-2) fucosylated glycans on C. albicans infection, an in vitro adhesion assay was performed with vaginal epithelial cells from wild type and Fut2-null mice. Vaginal epithelial cells from Fut2-null mice were found to bind increased numbers of C. albicans compared to vaginal epithelial cells obtained from wild type mice. Hysterectomy lessened the difference between Fut2-null and wild type mice in binding of C. ablicans in vitro and susceptibility to experimental C. albicans vaginitis in vivo. We generated a recombinant fucosylated MUC1 glycanpolymer to test whether the relative protection of wild type mice compared to Fut2-null mice could be mimicked with exogenous mucin. While a small portion of the recombinant MUC1 epitopes displayed α(1-2)fucosylated glycans, the predominant epitopes were sialylated due to endogenous sialyltransferases in the cultured cells. Intravaginal instillation of recombinant MUC1 glycanpolymer partially reduced experimental yeast vaginitis suggesting that a large glycanpolymer, with different glycan epitopes, may affect fungal burden.

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Abbreviations

CFU:

Colony forming unit

Fut2:

a(1,2)fucosyltransferase “Secretor” gene

UEA-I:

Ulex europaeus agglutinin I

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Acknowledgments

We thank Paul Fidel for the gift of C. albicans strain 3153A, the personnel of the Cancer Center Research Histology and Immunoperoxidase Core at the University of Michigan for histological processing, Janet Hoff at the Center for Integrative Genomics at the University of Michigan for performing abdominal oophorectomy, Brady West at the Centre for Statistical Consultation and Research of the University of Michigan, the Mammalian Protein Expression Core Facility, and Dr. Hasse Karlsson for assistance with the LC-MS analysis and spectra deduction. This work was supported by grants from the University of Michigan Biomedical Research Council and Horace H. Rackham School of Graduate Studies, the Swedish Research Council (equipment and grant no. 7461), and the IngaBritt and Arne Lundberg Foundation.

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Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Cellular and Molecular Biology Program, University of Michigan Medical Center, Ann Arbor, MI, 48109, USA

    Steven E. Domino, Elizabeth A. Hurd & David M. Karnak

  2. Department of Medical Biochemistry, University of Gothenburg, Medicinaregatan 9A, 413 90, Gothenburg, Sweden

    Kristina A. Thomsson, Jessica M. Holmén Larsson, Elisabeth Thomsson, Malin Bäckström & Gunnar C. Hansson

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  1. Steven E. Domino
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  2. Elizabeth A. Hurd
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  3. Kristina A. Thomsson
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  5. Jessica M. Holmén Larsson
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  6. Elisabeth Thomsson
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  8. Gunnar C. Hansson
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Correspondence to Steven E. Domino.

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Domino and Hurd contributed equally to this study.

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Supplemental Fig. 1

Vaginal epithelial cells obtained by vaginal lavage from wild type and Fut2-null mice stained with UEA-I and Alcian blue pH 2.5 with control panels shown. Wild type and Fut2-null mice underwent either control surgery (ovariectomy alone) or hysterectomy (ovariectomy and total hysterectomy) to remove the entire cervix along with uterus. Intense Alcian blue pH 2.5 mucin staining (top panel) was present in washings from ovariectomized control wild-type (A) and Fut2-null mice (C) but absent in washings from hysterectomized wild type (B) and Fut2-null mice (D). Duplicate slides stained with UEA-I (bottom panel) show specific brown lectin staining associated with vaginal epithelial cells from pseudoestrus ovariectomized wild type mice (E) but absent in washings from pseudoestrus ovariectomized Fut2-null (G), hysterectomized wild type (F) and Fut2-null (H) mice. Bar = 100 μm. (PDF 35.1 kb)

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Domino, S.E., Hurd, E.A., Thomsson, K.A. et al. Cervical mucins carry α(1,2)fucosylated glycans that partly protect from experimental vaginal candidiasis. Glycoconj J 26, 1125–1134 (2009). https://doi.org/10.1007/s10719-009-9234-0

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