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Herpes simplex virus-binding IgG traps HSV in human cervicovaginal mucus across the menstrual cycle and diverse vaginal microbial composition

Mucosal Immunologyvolume 11pages14771486 (2018) | Download Citation

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Abstract

IgG possesses an important yet little recognized effector function in mucus. IgG bound to viral surface can immobilize otherwise readily diffusive viruses to the mucin matrix, excluding them from contacting target cells and facilitating their elimination by natural mucus clearance mechanisms. Cervicovaginal mucus (CVM) is populated by a microbial community, and its viscoelastic and barrier properties can vary substantially not only across the menstrual cycle, but also in women with distinct microbiota. How these variations impact the “muco-trapping” effector function of IgGs remains poorly understood. Here we obtained multiple fresh, undiluted CVM specimens (n = 82 unique specimens) from six women over time, and employed high-resolution multiple particle tracking to quantify the mobility of fluorescent Herpes Simplex Viruses (HSV-1) in CVM treated with different HSV-1-binding IgG. The IgG trapping potency was then correlated to the menstrual cycle, and the vaginal microbial composition was determined by 16 s rRNA. In the specimens studied, both polyclonal and monoclonal HSV-1-binding IgG appeared to consistently and effectively trap HSV-1 in CVM obtained at different times of the menstrual cycle and containing a diverse spectrum of commensals, including G. vaginalis-dominant microbiota. Our findings underscore the potential broad utility of this “muco-trapping” effector function of IgG to reinforce the vaginal mucosal defense, and motivates further investigation of passive immunization of the vagina as a strategy to protect against vaginally transmitted infections.

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References

  1. 1.

    Gipson, I. Mucins of the human endocervix. Front. Biosci. 6, D1245–D1255 (2001).

  2. 2.

    Kieweg S. L. Gravity‐induced coating flows of vaginal gel formulations: In vitro experimental analysis J. Pharmaceut. Sci. 93, 2941–2952 (2014).

  3. 3.

    Nguyen, P. V. Innate and adaptive immune responses in male and female reproductive tracts in homeostasis and following HIV infection. Cell Mol. Immunol. 11, 410–427 (2014).

  4. 4.

    Cone, R. A. Barrier properties of mucus. Adv. Drug Deliv. Rev. 61, 75–85 (2009).

  5. 5.

    Cole, A. Innate host defense of human vaginal and cervical mucosae. Curr. Top. Microbiol Immunol. 306, 199–230 (2006).

  6. 6.

    Naz, R. K. Female genital tract immunity: distinct immunological challenges for vaccine development. J. Reprod. Immunol. 93, 1–8 (2012).

  7. 7.

    Kumamoto, Y. & Iwasaki, A. Unique features of antiviral immune system of the vaginal mucosa. Curr. Opin. Immunol. 24, 411–416 (2012).

  8. 8.

    Chen, A. et al. Transient antibody-mucin interactions produce a dynamic molecular shield against viral invasion. Biophys. J. 106, 2028–2036 (2014).

  9. 9.

    Wessler, T. et al. Using computational modeling to optimize the design of antibodies that trap viruses in mucus. ACS Infect. Dis. 2, 82–92 (2016).

  10. 10.

    Fahrbach, K. M., Malykhina, O., Stieh, D. J. & Hope, T. J. Differential binding of IgG and IgA to mucus of the female reproductive tract. PLoS One 8, e76176 (2013).

  11. 11.

    Wang, Y. Y. et al. Diffusion of immunoglobulin G in shed vaginal epithelial cells and in cell-free regions of human cervicovaginal mucus. PLoS One 11, e0158338 (2016).

  12. 12.

    Saltzman, W. M., Radomsky, M. L., Whaley, K. J. & Cone, R. A. Antibody diffusion in human cervical mucus. Biophys. J. 66(2 Pt 1), 508–515 (1994).

  13. 13.

    Olmsted, S. S. et al. Diffusion of macromolecules and virus-like particles in human cervical mucus. Biophys. J. 81, 1930–1937 (2001).

  14. 14.

    Wang, Y. Y. et al. IgG in cervicovaginal mucus traps HSV and prevents vaginal herpes infections. Mucosal Immunol. 7, 1036–1044 (2014).

  15. 15.

    Sharif K. The Structure, Chemistry and Physics of Human Cervical Mucus, Vol. 2nd edn. (Blackwell Publishing Ltd, Oxford, 2006).

  16. 16.

    Moncla, B. J., Chappell, C. A., Debo, B. M. & Meyn, L. A. The Effects of Hormones and Vaginal Microflora on the Glycome of the Female Genital Tract: Cervical-Vaginal Fluid. PLoS One 11, e0158687 (2016).

  17. 17.

    Chappell, C. A. et al. The effects of reproductive hormones on the physical properties of cervicovaginal fluid. Am. J. Obstet. Gynecol. 211, 226 e221–226 e227 (2014).

  18. 18.

    Aksoy, M., Guven, S., Tosun, I., Aydin, F. & Kart, C. The effect of ethinyl estradiol and drospirenone-containing oral contraceptives upon mucoprotein content of cervical mucus. Eur. J. Obstet. Gynecol. Reprod. Biol. 164, 40–43 (2012).

  19. 19.

    Shukair, S. A. et al. Human cervicovaginal mucus contains an activity that hinders HIV-1 movement. Mucosal Immunol. 6, 427–434 (2013).

  20. 20.

    Moncla, B. J. et al. Impact of bacterial vaginosis, as assessed by nugent criteria and hormonal status on glycosidases and lectin binding in cervicovaginal lavage samples. PLoS One 10, e0127091 (2015).

  21. 21.

    Nunn, K. L. et al. Enhanced Trapping of HIV-1 by Human Cervicovaginal Mucus Is Associated with Lactobacillus crispatus-Dominant Microbiota. MBio. 6, e01084–01015 (2015).

  22. 22.

    Gajer, P. et al. Temporal dynamics of the human vaginal microbiota. Sci. Transl. Med. 4, 132ra152 (2012).

  23. 23.

    Ravel, J. et al. Vaginal microbiome of reproductive-age women. Proc. Natl Acad. Sci. USA 108(Suppl 1), 4680–4687 (2011).

  24. 24.

    Ma, B., Forney, L. J. & Ravel, J. Vaginal microbiome: rethinking health and disease. Annu. Rev. Microbiol. 66, 371–389 (2012).

  25. 25.

    Borgdorff, H. et al. Cervicovaginal microbiome dysbiosis is associated with proteome changes related to alterations of the cervicovaginal mucosal barrier. Mucosal Immunol. 9, 621–633 (2016).

  26. 26.

    Lai, S. K. et al. Human Immunodeficiency Virus Type 1 Is Trapped by Acidic but Not by Neutralized Human Cervicovaginal Mucus. J. Virol. 83, 11196–11200 (2009).

  27. 27.

    Wira, C. R., Fahey, J. V., Rodriguez-Garcia, M., Shen, Z. & Patel, M. V. Regulation of mucosal immunity in the female reproductive tract: the role of sex hormones in immune protection against sexually transmitted pathogens. Am. J. Reprod. Immunol. 72, 236–258 (2014).

  28. 28.

    Lai, S. K., Wang, Y. Y., Hida, K., Cone, R. & Hanes, J. Nanoparticles reveal that human cervicovaginal mucus is riddled with pores larger than viruses. Proc. Natl Acad. Sci. USA 107, 598–603 (2010).

  29. 29.

    Whaley, K. J. & Zeitlin, L. Antibody-based concepts for multipurpose prevention technologies. Antivir. Res. 100(Suppl), S48–S53 (2013).

  30. 30.

    Zeitlin, L. et al. Enhanced potency of a fucose-free monoclonal antibody being developed as an Ebola virus immunoprotectant. Proc. Natl Acad. Sci. USA 108, 20690–20694 (2011).

  31. 31.

    Klipping, C., Duijkers, I., Trummer, D. & Marr, J. Suppression of ovarian activity with a drospirenone-containing oral contraceptive in a 24/4 regimen. Contraception 78, 16–25 (2008).

  32. 32.

    Han, L., Taub, R. & Jensen, J. T. Cervical mucus and contraception: what we know and what we don’t. Contraception 96, 310–321 (2017).

  33. 33.

    Clementi, N. et al. Role and potential therapeutic use of antibodies against herpetic infections. Clin. Microbiol. Infect. 23, 381–386 (2017).

  34. 34.

    Yamamoto, H. & Matano, T. Patterns of HIV/SIV Prevention and Control by Passive Antibody Immunization. Front Microbiol. 7, 1739 (2016).

  35. 35.

    Whaley, K. J. & Mayer, K. H. Strategies for preventing mucosal cell-associated HIV transmission. J. Infect. Dis. 210(Suppl 3), S674–S680 (2014).

  36. 36.

    Gosmann, C. et al. Lactobacillus-deficient cervicovaginal bacterial communities are associated with increased HIV acquisition in young south African women. Immunity 46, 29–37 (2017).

  37. 37.

    Wiggins, R. H. S., Soothill, P. W., Millar, M. R. & Corfield, A. P. Mucinases and sialidases: their role in the pathogenesis of sexually transmitted infections in the female genital tract. Sex. Transm. Inf. 77, 402–408 (2001).

  38. 38.

    Bertran, T. et al. Slight pro-inflammatory immunomodulation properties of dendritic cells by Gardnerella vaginalis: the “Invisible Man” of bacterial vaginosis? J. Immunol. Res. 2016, 9747480 (2016).

  39. 39.

    LeGoff, J., Pere, H. & Belec, L. Diagnosis of genital herpes simplex virus infection in the clinical laboratory. Virol. J. 11, 83 (2014).

  40. 40.

    Zeitlin, L. et al. Topically applied human recombinant monoclonal IgG1 antibody and its Fab and F(ab’)2 fragments protect mice from vaginal transmission of HSV-2. Virology 225, 213–215 (1996).

  41. 41.

    Fernandez-Romero, J. A. et al. Multipurpose prevention technologies: the future of HIV and STI protection. Trends Microbiol. 23, 429–436 (2015).

  42. 42.

    Uyangaa, E., Patil, A. M. & Eo, S. K. Prophylactic and therapeutic modulation of innate and adaptive immunity against mucosal infection of herpes simplex virus. Immune Netw. 14, 187–200 (2014).

  43. 43.

    Qiu, X. et al. Reversion of advanced Ebola virus disease in nonhuman primates with ZMapp. Nature 514, 47–53 (2014).

  44. 44.

    Roth, K., Ferreira, V. H. & Kaushic, C. HSV-2 vaccine: current state and insights into development of a vaccine that targets genital mucosal protection. Microb. Pathog. 58, 45–54 (2013).

  45. 45.

    Escolano, A., Dosenovic, P. & Nussenzweig, M. C. Progress toward active or passive HIV-1 vaccination. J. Exp. Med. 214, 3–16 (2017).

  46. 46.

    Kelley, B. Industrialization of mAb production technology. mAbs. 1, 443–452 (2009).

  47. 47.

    Liu, H. F., Ma, J., Winter, C. & Bayer, R. Recovery and purification process development for monoclonal antibody production. mAbs. 2, 480–499 (2014).

  48. 48.

    O’Hare, G. E. P. Live-cell analysis of a green fluorescent protein-tagged herpes simplex virus infection. J. Virol. 73, 4110–4119 (1999).

  49. 49.

    Apgar, J. et al. Multiple-particle tracking measurements of heterogeneities in solutions of actin filaments and actin bundles. Biophys. J. 79, 1095–1106 (2000).

  50. 50.

    Lai, Y. K. & Rosin, P. L. Efficient circular thresholding. IEEE Trans. Image Process 23, 992–1001 (2014).

  51. 51.

    Wang, Y. Y., Nunn, K. L., Harit, D., McKinley, S. A. & Lai, S. K. Minimizing biases associated with tracking analysis of submicron particles in heterogeneous biological fluids. J. Control Release. 220(Pt A), 37–43 (2015).

  52. 52.

    Fadrosh DW M. B., et al. An improved dual-indexing approach for multiplexed 16S rRNA gene sequencing on the Illumina MiSeq platform. Microbiome. 2, 6 (2014).

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Acknowledgements

This work was supported by the National Institutes of Health (http://www.nih.gov/) grants R21AI093242 (S.K.L.), U19AI096398 (S.K.L.), and U19AI084044 (J.R.), a Diversity Supplement 1F32AI102535 (K.L.N.), The David and Lucile Packard Foundation (https://www.packard.org/) 2013-39274 (S.K.L.), the Eshelman Institute of Innovation (http://unceii.org/, S.K.L.), and startup funds from the University of North Carolina Eshelman School of Pharmacy (https://pharmacy.unc.edu/; S.K.L). The funders had no role in study design, data collection and analysis, decision to publish, or manuscript preparation.

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Affiliations

  1. Division of Pharmacoengineering and Molecular Pharmaceutics, Eshelman School of Pharmacy, University of North Carolina-Chapel Hill, Chapel Hill, NC, 27519, USA

    • Holly A. Schroeder
    • , Kenetta L. Nunn
    • , Alison Schaefer
    • , Christine E. Henry
    • , Felix Lam
    •  & Samuel K. Lai
  2. UNC/NCSU Joint Department of Biomedical Engineering, University of North Carolina-Chapel Hill, Chapel Hill, NC, 27519, USA

    • Kenetta L. Nunn
    •  & Samuel K. Lai
  3. Mapp Biopharmaceutical Inc, San Diego, CA, 92121, USA

    • Michael H. Pauly
    • , Kevin J. Whaley
    •  & Larry Zeitlin
  4. Department of Microbiology and Immunology, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    • Mike S. Humphrys
    •  & Jacques Ravel
  5. Institute for Genome Sciences, University of Maryland School of Medicine, Baltimore, MD, 21201, USA

    • Jacques Ravel

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Contributions

H.A.S., K.L.N., and S.K.L. conceptualized and designed the experiments; K.L.N. recruited study participants; H.A.S., K.L.N., C.E.H., F.L., M.S.H., and S.K.L. performed experiments; H.A.S., A.S., and S.K.L. performed data analysis; M.H.L., K.J.W., and L.Z. provided reagents; H.A.S. and S.K.L. wrote the paper; H.A.S., K.J.W., L.Z., J.R., and S.K.L. edited the paper.

Competing interests

Intellectual property associated with harnessing antibody–mucin interactions described in part in this publication was developed at the University of North Carolina-Chapel Hill (UNC-CH), and has been licensed to Mucommune, L.LC. S.K.L. is a founder of Mucommune and currently serves as its interim CEO, board of director, and in the scientific advisory board. S.K.L. owns company stock; S.K.L.’s relationship with Mucommune is subject to certain restrictions under University policy. The terms of this arrangement are being managed by UNC-CH in accordance with its conflict of interest policies.

Corresponding author

Correspondence to Samuel K. Lai.

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DOI

https://doi.org/10.1038/s41385-018-0054-z