Ignite Creation Date:
2024-05-06 @ 6:58 PM
Last Modification Date:
2024-10-26 @ 2:58 PM
Study NCT ID:
NCT05846620
Status:
RECRUITING
Last Update Posted:
2024-01-24
First Post:
2023-04-27
Brief Title:
Pulmonary Immune Cell-microbiome Interactions in the Healthy Lung
Sponsor:
Hvidovre University Hospital
Organization:
Hvidovre University Hospital
Study Overview
Official Title:
Pulmonary Immune Cell-microbiome Interactions in the Healthy Lung The ILLUMINA-2 Study
Status:
RECRUITING
Status Verified Date:
2024-01
Last Known Status:
None
Delayed Posting:
No
If Stopped, Why?:
Not Stopped
Has Expanded Access:
False
If Expanded Access, NCT#:
N/A
Has Expanded Access, NCT# Status:
N/A
Acronym:
ILLUMINA-2
Brief Summary:
The overall aim is to to provide a normal material for the composition and spatial heterogeneity of the following in the healthy lung i immune cell populations and their activation patterns ii the surrounding cytokine-chemokine milieu including trans-compartmental fluxes of these mediators between the lung and bloodstream and iii the lung microbiome
Main hypotheses
Absolute and relative immune cell counts in bronchoalveolar lavage fluid BALF are similar to those previously reported by other methods67
No trans-compartmental flux of cytokines between the lungs and bloodstream is present but cytokine concentrations notably IL-6 and IL-8 vary with the immune-cell-microbiome composition
Immune cell mainly T cell activation differentiation and gene expression patterns are expected to differ between blood and BALF in a manner that depends on the regional diversity of the pulmonary microbiome
Main hypotheses
Absolute and relative immune cell counts in bronchoalveolar lavage fluid BALF are similar to those previously reported by other methods67
No trans-compartmental flux of cytokines between the lungs and bloodstream is present but cytokine concentrations notably IL-6 and IL-8 vary with the immune-cell-microbiome composition
Immune cell mainly T cell activation differentiation and gene expression patterns are expected to differ between blood and BALF in a manner that depends on the regional diversity of the pulmonary microbiome
Detailed Description:
The coronavirus disease 2019 pandemic caused by severe acute respiratory syndrome coronavirus 2 has brought with it several studies on the local pulmonary immune system which was until very recently largely a terra incognita With these new methods apparently fundamental aspects of lung disease have been uncovered including pulmonary hyper inflammation However very little is currently known about the composition of the normal immune cell population in the human lung including its interplay with the pulmonary microbiome that is the commensal microbiota of interacting bacteria and fungi that reside within the healthy lung
Overall design In 50 patients 25 males 25 females intubated for elective surgery in general anaesthesia an endotracheal aspirate and BAL fluid BALF from separate lung segments will be obtained Furthermore an oral and nasal swab and blood samples will be collected This will be done immediately after intubation
The following is obtained from the patients electronic health record diagnosis codes and medication type and dosage smoking history currentpreviousnever smoker pack years type of operation furthermore the health record is screened for any exclusion criteria
Blood samples are drawn from the patients peripheral venous catheter inserted for clinical purposes immediately before BALF collection
Bronchoscopy with BALF collection This procedure is performed in a standardized fashion according to current clinical guidelines Immediately prior to the procedure an oral swab nasal swab and an endotracheal aspirate ETA are obtained FIO2 is then increased to 10 and the bronchoscopy procedure is performed using a disposable videoscope with an outer diameter of 50 mm Three successive 50-ml aliquots of prewarmed 37C isotonic saline are instilled in the medial segment of the right middle lobe aspirated immediately with low negative suction pressure 100 cm H2O and pooled into a sterile glass container on ice to obtain a BALF specimen
Afterwards a mini-BAL is performed in the upper and lower lobe of the right lung with a single installation of 20 ml isotonic saline in each lobe with immediately aspiration into a sterile container
Measurements The composition of the immune cell population as well as the function and differentiation of various cell lines will be investigated by single-cell RNA sequencing scRNA-seq on selected immune cells from BALF ETA and blood and this will be supplemented by bulk RNA sequencing with sample barcoding and multiplexing giving a detailed expression pattern of all samples
The composition microbiome in BALF ETA and oral swabs will be assessed by targeted amplicon sequencing of the hyper-variable regions 1 through 3 of the 16S subunit of ribosomal RNA gene for bacteria
Statistical analyses will be performed using R statistical software version 411 R Project for Statistical Computing within RStudio statistical software version 141717 RStudio and p005 considered statistically significant Inspection of normality and variance homogeneity will be done by creating qq-plots and histograms The statistical inference tools SPIEC-EASI and HeatMaps will be used and based on correlational analyses principal component analyses including non-hierarchal cluster analysis will be applied to identify traits
Overall design In 50 patients 25 males 25 females intubated for elective surgery in general anaesthesia an endotracheal aspirate and BAL fluid BALF from separate lung segments will be obtained Furthermore an oral and nasal swab and blood samples will be collected This will be done immediately after intubation
The following is obtained from the patients electronic health record diagnosis codes and medication type and dosage smoking history currentpreviousnever smoker pack years type of operation furthermore the health record is screened for any exclusion criteria
Blood samples are drawn from the patients peripheral venous catheter inserted for clinical purposes immediately before BALF collection
Bronchoscopy with BALF collection This procedure is performed in a standardized fashion according to current clinical guidelines Immediately prior to the procedure an oral swab nasal swab and an endotracheal aspirate ETA are obtained FIO2 is then increased to 10 and the bronchoscopy procedure is performed using a disposable videoscope with an outer diameter of 50 mm Three successive 50-ml aliquots of prewarmed 37C isotonic saline are instilled in the medial segment of the right middle lobe aspirated immediately with low negative suction pressure 100 cm H2O and pooled into a sterile glass container on ice to obtain a BALF specimen
Afterwards a mini-BAL is performed in the upper and lower lobe of the right lung with a single installation of 20 ml isotonic saline in each lobe with immediately aspiration into a sterile container
Measurements The composition of the immune cell population as well as the function and differentiation of various cell lines will be investigated by single-cell RNA sequencing scRNA-seq on selected immune cells from BALF ETA and blood and this will be supplemented by bulk RNA sequencing with sample barcoding and multiplexing giving a detailed expression pattern of all samples
The composition microbiome in BALF ETA and oral swabs will be assessed by targeted amplicon sequencing of the hyper-variable regions 1 through 3 of the 16S subunit of ribosomal RNA gene for bacteria
Statistical analyses will be performed using R statistical software version 411 R Project for Statistical Computing within RStudio statistical software version 141717 RStudio and p005 considered statistically significant Inspection of normality and variance homogeneity will be done by creating qq-plots and histograms The statistical inference tools SPIEC-EASI and HeatMaps will be used and based on correlational analyses principal component analyses including non-hierarchal cluster analysis will be applied to identify traits
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
False
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
None
Is an FDA AA801 Violation?:
None