Investigating the cervicovaginal mucus barrier properties of women with bacterial vaginosis

Thuy Hoang

May 18, 2015

Highly adhesive and viscoelastic 3

Mucus coats entry points1 Nanoporous mesh2

1. Iwasaki, A., Nat Rev Immunol. 2010 2. Sanders et al., AJRCCM, 2000

3. Cone, R.A., Mucosal Immunology, 1999

Mucus is a barrier to pathogens

2

Mucus can sterically and adhesively block pathogens THE CENTER FOR NANOMEDICINE AT JOHNS HOPKINS

Cervicovaginal Mucus (CVM)

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• Mucus is secreted by endocervix

• Expelled into vaginal compartment through cervical os

• Mixes with: vaginal transudate, exfoliated epithelial cells, bacterial secretions

• In FRT, mucus lining is ~10 µm thick

• CVM is home to microbiota that can produce enzymes to modulate it

• Enzymes can degrade barrier properties of CVM

Iwasaki, A., Nat Rev Immunol. 2010

The vaginal microbiota varies among women

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• Lactobacilli dominated: -Near monomicrobial Lactobacilli -pH < 4.0 • Polymicrobial culture/Bacterial Vaginosis (BV): -Overgrowth of many different mostly gram-negative species -pH > 4.5 -Recurrence rate >50% within 12 months -BV incidence rate 29.2% in the US -BV increased risk of HIV acquisition by 60% -Risk factor for adverse obstetric outcomes, PID, acquisition/transmission of STIs

Lactobacillus-dominated

Polymicrobial/BV

Methods

Instead® menstrual cups

• Donor Mucus Undiluted CVM can be obtained from self-administered collection device • mCherry fluorescently labeled HIV

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• Quantitative Microscopy Technique -Multiple Particle Tracking (MPT) -Obtain x, y positional data over time -Calculate Mean-Square Displacement (MSD)

HIV is diffusive in polymicrobial CVM

Lactobacillus-dominated Polymicrobial/BV

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HIV is diffusive in polymicrobial CVM

10-3

10-2

10-1

100

101

0.01 0.1 1 10

Dis

tanc

e Tr

avel

ed<M

SD>

(µm

2 )

Time Scale (s)

Polymicrobial/BV

Water

Lactobacillus-dominated

Polymicrobial/BV

Lactobacillus-dominated

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HIV is diffusive in CVM even after antibiotic treatment

10-3

10-2

10-1

100

101

0.01 0.1 1 10

Dis

tanc

e Tr

avel

ed<M

SD>

(µm

2 )

Time Scale (s)

Before

Water

After

• BV is treated with course of antibiotics (Metronidazole, Clindamycin, Tinidazole)

• 1 month after treatment, HIV is still diffusive in CVM

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CVM barrier properties similar between women with symptomatic and asymptomatic BV

10-3

10-2

10-1

100

101

0.01 0.1 1 10

Dis

tanc

e Tr

avel

ed<M

SD>

(µm

2 )

Time Scale (s)

Symptomatic

Water

Asymptomatic

Lactobacillus-dominated

• Symptomatic - presented at a clinic for treatment

• Asymptomatic - self-reported “healthy” and without vaginal symptoms

• HIV diffusion was similar between these two groups

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HIV diffusion correlated with pH and lactic acid concentration

10-3

10-2

10-1

100

101

3 3.5 4 4.5 5 5.5 6 6.5 7

MSD

at 1

sec

ond

pH

Lactobacillus-dominated

Polymicrobial/BV

10-3

10-2

10-1

100

101

0 0.5 1 1.5 2M

SD a

t 1 s

econ

d% Lactic Acid

Lactobacillus-dominated

Polymicrobial/BV

• HIV diffusion correlated with higher pH (>4.5)

• Lactobacillus make lactic acid • Higher % lactic acid correlates with

HIV trapping

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Future Directions

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• Mechanisms by which BV-associated bacteria degrade CVM barrier properties

• Prospective clinical study to understand how

Depo-Provera may modulate the microbiota thus effecting the CVM barrier properties

Acknowledgements

Funding R01HD062844

Collaborators Richard Cone, Ph.D Kevin DeLong Sam Lai, Ph.D Craig Hendrix, MD Jenell Coleman Fennell, MD Ronald Schnaar, Ph.D

The Center for Nanomedicine Laura Ensign-Hodges, Ph.D Elizabeth Nance, Ph.D. Jane Chisholm Kunal Parikh Ben Schuster, Ph.D. Taarika Babu Yoo Chun Kim, Ph.D. Jong Sung Park, Ph.D. Rezo Omiadze, M.D.

Sumon Chattopadhyay Maggie Swierczewska, Ph.D. Tao Yu, Ph.D. Abhijit Date, Ph.D. Clark Zhang Raj Reddy, Ph.D. Xinglu Huang, Ph.D. Yi-An Lin, Ph.D. Emily Toler

Gregg Duncan, Ph.D. Heena Soni, Ph.D. Yumin Oh, Ph.D. Siva Kambhampati, Ph.D. Panos Mastorakos, M.D. Fan Zhang Uri Soiberman, M.D. Namho Kim Elga Bandeira de Melo Marta Roig Pons

Core faculty Justin Hanes, Ph.D. Rangaramanujam Kannan, Ph.D. Samuel Yiu, M.D. Ian Pitha, M.D., Ph.D. Jung Soo Suk, Ph.D. Qingguo Xu, Ph.D. Jie Fu, Ph.D. Seulki Lee, Ph.D. Manoj Mishra, Ph.D. Ogyi Park, Ph.D.

CVM donors

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