FEMALE SEX HORMONES INFLUENCE VAGINAL HIV 1 INFECTION AND - PowerPoint PPT Presentation

FEMALE SEX HORMONES INFLUENCE VAGINAL HIV ‐ 1 INFECTION AND DISSEMINATION IN A HUMANIZED MOUSE MODEL Philip V Nguyen, Jocelyn M Wessels, KristenMueller, Varun Anipindi, FatemahVahedi, Marianne Chew, Alexandre Deshiere, Tony Mazzulli, Michel J Tremblay, Ali A Ashkar, & Charu Kaushic Kristen Mueller McMaster University OHTN ENDGAME 1 Conference October 24, 2016

Presenter Disclosure • I have no conflicts of interest to declare

Women & HIV Approximately 40% of HIV transmission occurs in the female genital tract 12.6 million cases of HIV are estimated to be a result of vaginal intercourse In 2015 women account for 25% of new HIV cases in Ontario OHTN has identified women as one of their 5 priority populations most affected by HIV "This epidemic unfortunately remains Social & biological factors for increased infection an epidemic of women." rates Michel Sidibé, Executive Director of UNAIDS June 2010 United Nations News Centre

Biological Mechanism of HIV Infection Hormones Estrogen: Protective effects against viral infection Increase epithelial barrier integrity Decrease levels of HIV target cells Increase protease inhibitors Progesterone: Increased susceptibility to viral infections Epithelial barrier remodeling/disruption Immune cell recruitment Increased HIV target cells Hormonal Contraceptives Birse et al. J. Virol (Sep 2015).

Hormonal Contraception & HIV Risk Over 150 million women world wide use hormonal contraceptive Approximately 50 million women use the injectable synthetic progestin Depot medroxyprogesterone acetate (DMPA) Heffron et al 2011: associated DMPA use with ~2 ‐ fold increase in risk of HIV ‐ 1 acquisition & transmission

Hormonal Contraception & HIV Risk Research

Hormonal Contraception & HIV Risk Morrison CS, et al. (2015) PLoS Med 12(1): e1001778.

Early Events of HIV infection in Women HIV antibody detection Early Chronic Exposure In blood Infection Eclipse Phase Viral Expansion Virus Dissemination Reservoir established Research Aim: to study early events of HIV replication and spread in our model of heterosexual transmission using a humanized mouse model

Humanized Mouse model of HIV infection Humanization of Mice Reconstitution Irradiation Pups injected with 1x106 ‐ 2x106 Newborn pups exposed CD34 ‐ enriched human to sub ‐ lethal radiation hematopoietic stem cells Remove “mouse” Give mice “human” immune system immune system Endpoint: Blood 1 ‐ 12 Weeks collection, Harvest Tissues Monitor Mice, Flow Cytometry, Blood collection, Histology, PCR Vaginal washes HIV ‐ 1 12 Week old Hu ‐ Mice Intravaginal Inoculation Reconstitution level confirmed in 10 3 ‐ 10 5 TCID50/mL of circulating PBMCs NL4.3 ‐ Bal ‐ Env HIV ‐ 1

HIV ‐ 1 viral burden declines in vaginal lavage and increases in plasma following intravaginal infection Plasma Vaginal Wash * Vaginal lavage HIV -1 RNA (copies/mL) ** * Plasma HIV -1 RNA (copies/mL) * ** Time post-inoculation (weeks) Time post-inoculation (weeks)

Dissemination of HIV ‐ 1 following intravaginal infection of humanized mice 5 weeks post infection – 10 3 1 week post infection – 10 3 HIV -1 RNA (copies/mL) HIV -1 RNA (copies/mL)

Intravaginal HIV ‐ 1 infection in humanized mice leads to a decrease in circulating human CD4 T cells over time 100 1 Week * 80 hCD4 (%) 60 40 20 0 Initial Endpoint 100 12 Weeks * 80 hCD4 (%) 60 * 40 20 0 Initial Endpoint

Intravaginal model of HIV infection in Humanized Mice Following intravaginal exposure HIV establishes as a local infection in the vaginal tract of humanized mice that disseminates systemically to other tissues within the body within 3 ‐ 5 weeks Humanized mice provide a good model to understand heterosexual HIV transmission in women

The effects of sex hormones on intravaginal HIV infection in a humanized mouse model 1) Examine if different stages of the female reproductive cycle had any effect on susceptibility of humanized ‐ mice to HIV ‐ 1 2) Compare susceptibility of hu ‐ mice following DMPA administration Endpoint: Blood Estrus (Estrogen High) 1 ‐ 5Weeks collection, Harvest Vaginal wash Diestrus (Progesterone High) Tissues for Estrus Monitor Mice, Flow Cytometry, Staging Blood collection, DMPA Injection Histology, PCR Vaginal washes HIV ‐ 1 12 Week old Hu ‐ Mice Intravaginal Inoculation Reconstitution level 10 3 ‐ 10 5 TCID50/mL of confirmed in circulating PBMCs NL4.3 ‐ Bal ‐ Env HIV ‐ 1

Hormonal environment in mice determines infection rate in intravaginal HIV exposure 10 6 10 5 10 4 Percent (%) 10 3 85% 0% 68% 10 2 10 1 10 0 Estrus Diestrus DMPA

HIV ‐ 1 levels in plasma is comparable between Diestrus and DMPA treated mice 1 Week Post ‐ Challenge 3 Weeks Post ‐ Challenge 5 Weeks Post ‐ Challenge Plasma HIV ‐ 1 RNA (copies/mL) P=0.1979 P=0.5826 P=0.3993

Dissemination of HIV ‐ 1 following intravaginal infection of Diestrus and DMPA treated mice

Uninfected DMPA treated humanized mice have increased levels of Target cells in blood 100 100 80 80 * * 60 60 40 40 20 20 0 0 Diestrus DMPA Diestrus DMPA

Summary Estrogen Progesterone DMPA High High treated 1.4 fold increase risk of Studies suggest Studies suggest HIV acquisition protection against viral susceptible period to viral infections infections We see comparable HIV levels & viral spread as Progesterone High mice Our mice model doesn’t We see local infection & We see increased get infected then systemic spread infection rates & target cells in blood

Acknowledgements Kaushic Lab Collaborators Dr. Charu Kaushic Ashkar Lab Dr. Jocelyn Wessels Dr. Ali Ashkar Philip Nguyen FatemahVahedi Kristen Mueller Marianne Chew Dr. Atif Zahoor Dr. Matthew Woods Dr. Michel Tremblay Puja Bagri Alexandre Deshiere Sara Dizzell Danielle Vitali Mt. Sinai Hospital & Jeff Lam University Health Network Maeve Cooper Dr. Tony Mazzulli Yung Lee John Ng, Ana Aquino Nishant Heryani Padmaja Sreeram Canadian HIV Vaccine Initiative www.chvi-icvv.gc.ca

Hladik, F. & McElrath, M. J. Setting the stage: host invasion by HIV. Nature Reviews Immunology 8 , 447 ‐ 457 (2008).

Follicular Phase: Estrogen Dominant  Protease inhibitors  Epithelial barrier integrity (cell ‐ cell adhesion factors)  Levels of cornification differentiation factors  Amounts of proximal HIV target cells Less hospitable for HIV infection? Luteal Phase: Progesterone Dominant  Protease mediated epithelial barrier remodeling/disruption  Immune cell recruitment  HIV target cells Ideal conditions for HIV infection? Birse et al. J. Virol (Sep 2015).

Net change in maternal & HIV ‐ related deaths from removing all injectable contraceptives WHO recommends women to discuss HIV risk and DMPA use with their doctor to determine if it is a suitable choice of contraceptive DMPA users recommended to use barrier methods to prevent STI transmission Insufficient evidence to globally condemn DMPA use

Expression of viral p24 ‐ antigen begins in the vaginal tract before spreading systemically Vagina Blood Spleen Week 1 Week 1 Week 1 p24 p24 p24 Spleen Vagina Blood Week 5 Week 5 Week 5 p24 p24 p24

HIV ‐ 1 infection is independent of the level of human CD45+ cells in the peripheral blood d b c Low-Med Blood Reconstitution (4.1% hCD45+) HIV -1 RNA (copies/mL) f g e High Blood Reconstitution (25.2% hCD45+) Plasma Vaginal lavage

Dissemination of HIV ‐ 1 in NRG mice following intravaginal infection **Based on 4 th mouse 7/18 positive tissues….waiting for Toronto results – may change!

HIV ‐ 1 levels in vaginal lavages lower in DMPA treated mice compared to Diestrus mice 1 Week Post ‐ Challenge 3 Weeks Post ‐ Challenge 5 Weeks Post ‐ Challenge Vaginal lavage HIV ‐ 1 RNA (copies/mL) *** * P=0.3962 P=0.0005 P=0.0420

10 5 Infectious Units 10 3 Infectious Units A B Vaginal lavage HIV ‐ 1 RNA Vaginal lavage HIV ‐ 1 RNA (copies/mL) (copies/mL) Time post ‐ challenge (weeks) Time post ‐ challenge (weeks) D C 10 5 Infectious Units 10 3 Infectious Units Plasma HIV ‐ 1 RNA Plasma HIV ‐ 1 RNA (copies/mL) (copies/mL) Time post ‐ challenge (weeks) Time post ‐ challenge (weeks)