Investigating the cervicovaginal mucus barrier
properties of women with bacterial vaginosis
Thuy Hoang
May 18, 2015
Mucus is a barrier to pathogens
Mucus coats entry
points1
Nanoporous mesh2
Highly adhesive and viscoelastic 3
Mucus can sterically and adhesively block pathogens
2
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1. Iwasaki, A., Nat Rev Immunol. 2010
2. Sanders et al., AJRCCM, 2000
3. Cone, R.A., Mucosal Immunology, 1999
Cervicovaginal Mucus (CVM)
• 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
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Iwasaki, A., Nat Rev Immunol. 2010
The vaginal microbiota varies among women
• Lactobacilli dominated:
-Near monomicrobial Lactobacilli
-pH < 4.0
Lactobacillus-dominated
• 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
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Polymicrobial/BV
Methods
• Donor Mucus
Undiluted CVM can be obtained from self-administered
collection device
• mCherry fluorescently labeled HIV
Instead® menstrual cups
• Quantitative Microscopy Technique
-Multiple Particle Tracking (MPT)
-Obtain x, y positional data over time
-Calculate Mean-Square Displacement (MSD)
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HIV is diffusive in polymicrobial CVM
Lactobacillus-dominated
Polymicrobial/BV
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HIV is diffusive in polymicrobial CVM
Polymicrobial/BV
Lactobacillus-dominated
Water
0
10
2
<MSD> (µm )
Distance Traveled
1
10
Polymicrobial/BV
-1
10
-2
10
Lactobacillus-dominated
-3
10
0.01
0.1
1
Time Scale (s)
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10
HIV is diffusive in CVM even after antibiotic
treatment
Water
0
10
2
<MSD> (µm )
Distance Traveled
1
10
Before
After
-1
10
-2
10
-3
10
0.01
0.1
1
Time Scale (s)
10
• 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
1
10
0
10
Symptomatic
Asymptomatic
2
<MSD> (µm )
Distance Traveled
Water
-1
10
-2
10
Lactobacillus-dominated
-3
10
0.01
0.1
1
10
Time Scale (s)
• 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
1
1
10
10
Polymicrobial/BV
0
10
MSD at 1 second
MSD at 1 second
Polymicrobial/BV
-1
10
0
10
-1
10
-2
10
-2
Lactobacillus-dominated
10
Lactobacillus-dominated
-3
10
-3
3
3.5
4
4.5
5
5.5
6
6.5
7
10
0
1
1.5
2
% Lactic Acid
pH
• HIV diffusion correlated with higher pH
(>4.5)
0.5
• Lactobacillus make lactic acid
• Higher % lactic acid correlates with
HIV trapping
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Future Directions
• 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
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Acknowledgements
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.
Funding
R01HD062844
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
CVM donors
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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
Collaborators
Richard Cone, Ph.D
Kevin DeLong
Sam Lai, Ph.D
Craig Hendrix, MD
Jenell Coleman Fennell, MD
Ronald Schnaar, Ph.D