Inducing Immune Quiescence to Prevent HIV Infection in Women

Study Overview
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Brief Summary
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In this project, the investigators want to analyse the capacity of Acetylsalicylic acid and hydoxychlroquin (HCQ) to induce an Immune Quiescence (IQ) phenotype, which has been previously associated with natural protection to HIV infection. This phenotype is characterized by lower expression of genes involved in cellular activation, lower resting levels of inflammatory cytokine production, lower level of systemic activated T cells, increased levels of systemic T regulatory, increased production of anti-viral anti-protease serpins at the female genital tract and reduced numbers of HIV target cells (mainly CD4+ CCR5+ T cells) in the FGT ( female genital tract). The objective of this study is to determine if daily oral administration of Acetylsalicylic acid or hydroxychlroroquin can reduce systemic and mucosal immune activation in HIV negative women.

Detailed Description
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The investigators will enrol 80 non female sex work low-risk HIV negative women and 80 HIV negative female sex worker HIV negative form Nairobi, Kenya and followed for a 3 months period. During the first month, samples will be taken on a monthly basis blood, vaginal samples: cytobrush/scarper and cervico vaginal lavage (CVL). This is done to determine the baseline levels of systemic and mucosal immune activation of each woman. In this way, every women is acting as her own control thereby reducing variation between control and participant. Chemokine/cytokine level, as well as cellular immune activation and T regulatory cells will be assessed. At month two: the women will be divided in two different arms (oral administration of hydroxychloroquine: 200mg/day or Acetylsalicylic acid 81mg/day) and followed, on a monthly basis, for an 8 additional weeks. During this time, monthly blood and vaginal samples (cytobrush/scraper and CVL) the investigators will be taken. They will measure change in the systemic and mucosal immune activation. Immune Quiescence phenotype (decrease of T cells immune activation, lower immune genes activation expression and pro-inflammatory cytokine/chemokine expression) will be evaluated by flow cytometry, microarray, and multiplex bead array technology. Here is how samples will be taken: A sample of cervical mucus will be collected by using a cotton swab rotated 360º in the cervical os, and a second swab used to collect secretions from the posterior vaginal fornix. Both swabs will be transferred into a single vial containing 5 mL of phosphate-buffered saline (PBS) which will be transported to the laboratory to be tested and cultured for sexually transmitted infections such as gonorrhea, chlamydia etc. Cervical cells will be collected by using a small brush and a wooden spatula. Both specimen will be transferred into a 15ml conical tube containing 5 ml of PBS. This specimen will be used to characterize the cellular populations in the specimen. Cervico vaginal lavage will be performed by washing the endocervix with 2 ml of sterile 1x PBS. The liquid will be collected form the posterior fornix. Samples will be placed into a conical tube, centrifuged to remove cellular debris and the supernatant will be stored at -70°C and will be shipped in liquid nitrogen dry shipper to Winnipeg, Manitoba. Those specimens will be used for innate soluble factor detection (chemokines, cytokines, antibodies, innate protein, etc 30ml of venous blood will be taken. (Peripheral Blood Mononuclear Cells will be extracted for immune activation analysis, DNA will be used for immune genes expression, plasma will be used for protein and innate soluble factor detection.) .

Official Title
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Limiting HIV Target Cells by Inducing Immune Quiescence in the Female Genital Tract

Conditions
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HIV

Intervention / Treatment
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* Drug: Acetylsalicylic Acid (ASA)
* Drug: Hydroxychloroquine (HCQ)


Participation Criteria
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Eligibility Criteria
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Inclusion criteria: Age greater than 18 years old and less than 50 years old Uterus and cervix present Willing to take daily acetylsalicylic acid or HCQ Willing to undergo pelvic exams In general good health, no chronic infection and not taking any anti-inflammatory or immunosuppressors Being HIV negative Without any cardiovascular disease Being active in sex work (for the Female commercial sex worker group) Exclusion criteria: Age less than 18 years or more than 50 years old Pregnancy (if a women becomes pregnant during the 10 weeks of the project she will be excluded) Breast feeding Pregnant in the last 12 months Being positive for Sexual transmissible disease or bacterial vaginosis at week 0 Menopausal No longer involve in sex work (for the female sex worker group) Having a chronic disease Taking any of the medication listed in annex 1 for health conditions Being allergic to acetylsalicylic acid, other medication for pain or fever, tartrazine dye or chloroquine, hydroxuchloroquine, primaquine or any other medication Having heartburn, stomach pain, stomach ulcer, anemia, hemophilia, kidney or liver disease, psoriasis, porphyria or other blood disease, G-6-PD deficiency, dermatitis (skin inflammation), alcoholism Having experienced previous vision changes while taking chloroquine, hydroxychloroquine (Aralen) or primaquine. Having a history of a diagnosed cardiovascular event, heart failure, peripheral arterial disease, angina, stoke, transient ischemic attack Having a current or recurrent condition with a high risk of major bleeding Having anemia Current participation in a clinical trial - -

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 50 Years

Sexes Eligible for Study
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Female

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Other: Acetylsalicylic Acid (ASA)<br>ASA 81 mg. p.o. daily for two months | Drug: Acetylsalicylic Acid (ASA)<br>* Acetylsalicylic Acid (ASA) 81 mg. oral daily for two months<br>|
| Other: Hydroxychloroquine (HCQ)<br>Hydroxychloroquine (HCQ) 200 mg. o.d. p.o. for two months. | Drug: Hydroxychloroquine (HCQ)<br>* Hydroxychloroquine (HCQ) 200 mg. oral, daily for two months.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in Systemic Immune Activation From Baseline Observed by the CD69 Expression on CD4 T Cells | We will analyse reduce of immune activation by measuring change in T cell activation (CD69) between baseline and every month during drug administration phase (8 weeks). | Baseline and 8 weeks | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in Number of CCR5+CD4+ T Cell Population at the Female Genital Tract. | We will measure changes in the number of CD4+T cells expressing CCR5 at the female genital tract before and at the end of the study. | baseline and 8 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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HIV, Immune quiescence, ASA, Hydroxychloroquin