Impact of Smoking and Its Cessation on Systemic and Airway Immune Activation
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|ClinicalTrials.gov Identifier: NCT02836067|
Recruitment Status : Recruiting
First Posted : July 18, 2016
Last Update Posted : August 14, 2019
|Condition or disease||Intervention/treatment||Phase|
|HIV Smoking||Behavioral: Counseling Drug: Smoking Cessation drugs Procedure: Bronchoscopy Procedure: Blood Draw Behavioral: Questionnaires||Not Applicable|
For Aim 1, 20 patients with HIV disease who have never smoked will be recruited with comparison to 20 active smokers with HIV disease, without evidence of COPD from spirometry, who are matched in demographics. Smokers who are interested in participating in a smoking cessation program will be referred to our Clinical Trials Unit (CTU) for all subsequent study visits.
Additionally, a comparison group of 20 uninfected smokers who have already been enrolled in the co-investigator's (Dr. Kwon) study will be used as comparison group. These patients have similar inclusion/exclusion criteria as this study and have been verified to be HIV-antibody negative. We will obtain de-identified samples and immunological and virological data already collected by Dr. Kwon. De-identified banked PBMC, plasma and BAL samples will also be accessible to us using a material transfer agreement to perform epithelial transcriptional gene expression profiling.
For Aim 2, a total of 100 HIV-infected individuals on effective ART who are active smokers and interested in participating in a smoking cessation program will be recruited. If 20 individuals who achieve 10-week of cessation are enrolled before 100 HIV smokers are fully enrolled, enrollment will cease, as 100 participants is an overestimate of the number of patients needed need to enroll to have 20 subjects achieve successful smoking cessation. The maximum total number of patients needed for the grant is 120 (100 HIV smokers, 20 HIV non-smokers).
We will recruit up to 120 patients from the BMC Center for Infectious Diseases (CID) outpatient clinic. The clinic serves the largest HIV-infected population in Boston, approximately 1,700 persons, and is composed largely of an urban socioeconomically disadvantaged population. Over 50% of HIV-infected patients in the CID are smokers, and >60% (based on prescription history of NRT, bupropion, varenicline) have attempted smoking cessation. We will recruit from flyers, the BMC ReSPECT registry, medical record screening, and physician referrals. A listing of the trial will also be posted on clinicaltrials.gov.
|Study Type :||Interventional (Clinical Trial)|
|Estimated Enrollment :||120 participants|
|Intervention Model:||Parallel Assignment|
|Masking:||None (Open Label)|
|Primary Purpose:||Basic Science|
|Official Title:||Impact of Smoking and Its Cessation on Systemic and Airway Immune Activation|
|Actual Study Start Date :||June 15, 2017|
|Estimated Primary Completion Date :||September 2021|
|Estimated Study Completion Date :||September 2021|
HIV-positive smokers will be enrolled in a smoking cessation program including the following procedures:
Counseling Smoking cessation drugs Questionnaires Blood Draw Bronchoscopy
Smoking cessation counseling
Drug: Smoking Cessation drugs
Medication to aid smoking cessation
Procedure: Blood Draw
Screening and research blood draws
HIV-positive non-smokers will be enrolled as a comparison group to HIV-positive smokers and will have the following procedures:
Questionnaires Blood Draw Bronchoscopy
Procedure: Blood Draw
Screening and research blood draws
- .Difference in T cell and monocyte immune subsets, level of activation [ Time Frame: Week 12 ]Aliquots of BAL and PBMC will be stained for surface antibodies to distinguish differentiation and activation markers. Monocyte cells will be phenotyped by CD3, CD19 and CD56 all on FITC, CD14 BUV 395, CD16 BV510, CCR2 PE, CX3CR1 APC, CD11c PEcf594, CD80 BV 421, CD86 BV 605, HLA-DR BV785, CD123 PE Cy7 and eFluor780 fixable viability dye. T cell populations will be stained for CD3, CD4, CD8, CD45RA, CCR7, CD27 and CD28; activation status by expression of CD25, CD38, CD69, HLA-DR, OX40. We will determine if they are Th2/Tc2-type cells by expression of CCR4, CCR8, T1/ST2, CRTH2. Flow will be performed on a LSRII (BD) and analyzed with FlowJo (Tree Star).
- Difference in levels of plasma inflammatory markers [ Time Frame: Week 12 ]For analysis of inflammatory protein levels, BAL fluid will be concentrated 10-fold using a Centricon filter (Millipore) with a 3,000 MW cutoff. We have found that assaying for cytokines is more reliable when the BAL is concentrated 10-fold since BAL is diluted ~100-fold by the procedure. sCD14 (R&D) and iFABP (Hycult) will be measured by ELISA. LPS will be measured with the LAL assay (Pierce) and sCD163 (Trillium Diag). Additional human cytokines associated with inflammatory processes, including TNF-α, IFN-α/g, IL-6/7/10, IP-10 and MCP-1, will be measured using the Milliplex cytokine- kit (Millipore), read on a Luminex 100 and analyzed with Beadview software (Upstate Cell).
- Differences in level of oxidative stress [ Time Frame: Week 12 ]Cells from blood and BAL will be stimulated with and without PMA for 30 min. and incubated with DHR123 to directly detect intracellular ROS production. Cells will be stained with fluorescent antibodies recognizing lineage markers for T cells (CD3, CD4, CD8), macrophages (CD14), B cells (CD19, CD20), and granulocytes (CD66b).
- Differences in the level of measure of HIV residual viremia [ Time Frame: Week 12 ]Measure levels of HIV-1 ca-DNA, ca-RNA and 2-LTR circles in BAL and PBMC samples collected from HIV-infected smokers versus non-smokers. Cellular DNA and RNA from PBMCs and BAL cells will be extracted with AllPrep DNA/RNA mini kit (Qiagen). HIV-1 ca-DNA will be quantified using a sensitive quantitative PCR (qPCR) assay to measure a conserved LTR/gag region96, and modified in our lab118. Measurements of ca-DNA will be reported as copies of HIV DNA/106 cells. Quantification of a conserved region of the human CCR5 will be used to determine the number of cells in each sample well.
- Differences airway transcriptional profile [ Time Frame: Week 12 ]Compare gene expression profiles between HIV smokers and non-smokers. Library preparation for RNA sequencing will be accomplished using Illumina's TruSeq RNA Sample Prep Kit v2, using 200-500ng of total RNA from each bronchial epithelial brushing. Briefly, RNA will be isolated using poly(A) selection, fragmented into a range of lengths centered around 200 base pairs, and randomly primed for reverse transcription followed by first and second-strand synthesis to create double-stranded cDNA fragments. Subsequently, these fragments will undergo PCR amplification, purification, and be used for cluster generation on a cBot machine using Illumina TruSeq Paired-End Cluster Generation Kits.
Please refer to this study by its ClinicalTrials.gov identifier (NCT number): NCT02836067
|Contact: Nina Lin, M.D.||email@example.com|
|Contact: Jennifer Bombardfirstname.lastname@example.org|
|United States, Massachusetts|
|Boston Medical Center||Recruiting|
|Boston, Massachusetts, United States, 02118|
|Contact: Nina Lin, MD 617-414-5242 email@example.com|
|Principal Investigator:||Nina Lin, M.D.||Boston Medical Center|