Cystic Fibrosis
Conditions
Keywords
cystic fibrosis, IL-17 neutrophils
Brief summary
The purpose of this study is to determine whether IL-17 polymorphonuclear leukocytes (PMNs) are central to the disease pathology in CF. This will be determined by demonstrating that IL-17 PMNs are present in the CF airway, correlate with lung function measures, and decrease in patients being treated with IV antibiotics for a pulmonary exacerbation.
Detailed description
This study consists of two parts which will be conducted in parallel. In the first part of the study, 14 subjects will be recruited for the Clinically Stable Cohort. Subjects will be asked to provide 1 gm of expectorated sputum and 40-ml of blood. A cell count and differential will be performed on the sputum followed by analysis for IL-17 PMNs by fluorescence-activated cell sorter (FACS). IL-17 PMNs also will be isolated by running the remainder of the cell pellet through a column of magnetic beads designed for this purpose. These neutrophils will be lysed and intracellular cytokines determined. Sputum supernatants will be stored frozen until analyzed for the presence of IL-1β, IL-6, IL-8, IL-17A, TGF-β, TNF-α, and neutrophil elastase. Clinical data will be captured from the subject's clinical outpatient visit including lung function measures and clinical culture results. IL-17 PMNs will be correlated with lung function measures and inflammatory mediators at baseline. In the second part of this study, 10 subjects will be recruited for the Exacerbation Cohort. Subjects will provide at least 1 gram of expectorated sputum and 40-ml of blood within 72 hours of hospital admission. Sputum and blood will be processed and analyzed as described above. Sputum and blood also will be obtained at the end of treatment (within 72 hours of completion of IV antibiotics) when the subject is at his or her baseline as determined by the managing physician. Clinical data will be captured from the subject's hospitalization records including lung function measures and clinical culture results.
Interventions
None listed
Sponsors
Case Western Reserve University
University Hospitals Cleveland Medical Center
Study design
Observational model
CASE_ONLY
Time perspective
PROSPECTIVE
Eligibility
Sex/Gender
ALL
Age
18 Years to 50 Years
Healthy volunteers
No
Inclusion criteria
* For Both Cohorts: ≥ 18 \< 50 years of age * For Both Cohorts: Must have a documented diagnosis of CF (positive sweat chloride ≥60 milliequivalents (mEq)/liter, by pilocarpine iontophoresis) and/or a genotype with two identifiable mutations consistent with CF accompanied by one or more clinical features with the CF) phenotype * For Both Cohorts: Chronically infected with P. aeruginosa defined by 2 positive cultures in the past year or 3 in the last two years * For Both Cohorts: Ability to expectorate mucus * For Both Cohorts: Ability to provide written informed consent * For Clinically Stable Cohort: 4 subjects with one copy of G551D * For Clinically Stable Cohort: 10 subjects who do not have G551D, they must have one copy of F508del * For Clinically Stable Cohort: CF subjects must have a baseline FEV1 percent predicted \> 50% (in the last year, obtained from medical record) * For Clinically Stable Cohort: Clinically stable: free of any acute illness for \>14 days * For Clinically Stable Cohort: Must have performed spirometry for clinical purposes at that clinical visit * For Clinically Stable Cohort: Must have a sputum culture sent to the clinical lab for clinical purposes at the time of the study visit * For Clinically Stable Cohort: Have not been prescribed any new systemic antibiotics for the 14 days prior to enrollment * For Exacerbation/IV Antibiotics Cohort: One copy of F508del * For Exacerbation/IV Antibiotics Cohort: Must have a doctor defined pulmonary exacerbation requiring treatment with IV antibiotics * For Exacerbation/IV Antibiotics Cohort: Must have performed spirometry for clinical purposes within 72 hours of initiating IV antibiotics and within 72 hours of completing IV antibiotics * For Exacerbation/IV Antibiotics Cohort: Must have a sputum culture sent to the clinical lab for clinical purposes within 72 hours of admission
Exclusion criteria
* For Both Cohorts: Pregnancy (based on self-report) * For Both Cohorts: Co-infection with Burkholderia cepacia complex organisms * For Both Cohorts: Any condition that in the opinion of the subject or the subject's managing physician that would compromise that individuals ability to participate in these studies * For Both Cohorts: Inability to tolerate the study procedures
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Change in Sputum IL-17 Neutrophils | End of Treatment, two weeks. Samples will be obtained from each study volunteer at the beginning of IV antibiotic treatment and at the completion of antibiotic treatment for a pulmonary exacerbation | In the Exacerbation/IV Antibiotics Cohort--Subjects will serve as their own controls. The percentage of neutrophils (in sputum) positive for IL-17 was determined by flow cytometry for each subject at the beginning and end of treatment for a pulmonary exacerbation. Sputum IL-17 neutrophil counts will be compared to the change in lung function (FEV1) as determined by spirometry (American Thoracic Society standards). |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, TGF-β, TNF-α, and Neutrophil Elastase | Samples will be obtained at one outpatient clinic visit during the next calendar year | In the Clinically Stable Cohort--Measurement of sputum inflammatory mediators: IL-1β, IL-6, IL-8, IL-17A, TGF-β, TNF-α, and neutrophil elastase |
| Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | End of Treatment, two weeks. Samples will be obtained from each study volunteer at the beginning of IV antibiotic treatment and at the completion of antibiotic treatment for a pulmonary exacerbation | In the Exacerbation/IV Antibiotics Cohort--Measurement of sputum inflammatory mediators by multiplex assay for IL-1β, IL-6, IL-8, and IL-17A. Neutrophil elastase determined by colorimetric assay. Measurements at the beginning of IV antibiotic treatment and after 2 weeks antibiotic treatment for a pulmonary exacerbation |
Countries
United States
Participant flow
Participants by arm
| Arm | Count |
|---|---|
| Clinically Stable Patients with cystic fibrosis and are clinically stable, 10 subjects with one copy of F508del and 4 subjects with at least one copy of G551D | 3 |
| Exacerbation Patients with cystic fibrosis admitted for treatment of a pulmonary exacerbation with IV antibiotics, 10 subjects with one copy of F508del | 11 |
| Total | 14 |
Baseline characteristics
| Characteristic | Clinically Stable | Exacerbation | Total |
|---|---|---|---|
| Age, Categorical <=18 years | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical >=65 years | 0 Participants | 0 Participants | 0 Participants |
| Age, Categorical Between 18 and 65 years | 3 Participants | 11 Participants | 14 Participants |
| Region of Enrollment United States | 3 participants | 11 participants | 14 participants |
| Sex: Female, Male Female | 1 Participants | 6 Participants | 7 Participants |
| Sex: Female, Male Male | 2 Participants | 5 Participants | 7 Participants |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk |
|---|---|---|
| deaths Total, all-cause mortality | — / — | — / — |
| other Total, other adverse events | 0 / 3 | 0 / 11 |
| serious Total, serious adverse events | 0 / 3 | 0 / 11 |
Outcome results
Primary
Change in Sputum IL-17 Neutrophils
In the Exacerbation/IV Antibiotics Cohort--Subjects will serve as their own controls. The percentage of neutrophils (in sputum) positive for IL-17 was determined by flow cytometry for each subject at the beginning and end of treatment for a pulmonary exacerbation. Sputum IL-17 neutrophil counts will be compared to the change in lung function (FEV1) as determined by spirometry (American Thoracic Society standards).
Time frame: End of Treatment, two weeks. Samples will be obtained from each study volunteer at the beginning of IV antibiotic treatment and at the completion of antibiotic treatment for a pulmonary exacerbation
Population: Regarding Clinically Stable Arm: Because very few neutrophils at the end of treatment for a pulmonary exacerbation were positive for IL-17, it was determined not to undertake studies examining sputum neutrophils during periods of clinical stability.
| Arm | Measure | Value (MEAN) | Dispersion |
|---|---|---|---|
| Begining of Exacerbation | Change in Sputum IL-17 Neutrophils | 55 % of neutrophils positive for IL-17 | Standard Deviation 23 |
| End of Exacerbation | Change in Sputum IL-17 Neutrophils | 4 % of neutrophils positive for IL-17 | Standard Deviation 9 |
Secondary
Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase
In the Exacerbation/IV Antibiotics Cohort--Measurement of sputum inflammatory mediators by multiplex assay for IL-1β, IL-6, IL-8, and IL-17A. Neutrophil elastase determined by colorimetric assay. Measurements at the beginning of IV antibiotic treatment and after 2 weeks antibiotic treatment for a pulmonary exacerbation
Time frame: End of Treatment, two weeks. Samples will be obtained from each study volunteer at the beginning of IV antibiotic treatment and at the completion of antibiotic treatment for a pulmonary exacerbation
| Arm | Measure | Group | Value (MEAN) | Dispersion |
|---|---|---|---|---|
| Begining of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-1b (pg/ml) | 2313 pg/ml | Standard Deviation 1452 |
| Begining of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-17 (pg/ml) | 302 pg/ml | Standard Deviation 185 |
| Begining of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-6 (pg/ml) | 53 pg/ml | Standard Deviation 26 |
| Begining of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | Neutrophil elastase activity (pg/ml) | 195 pg/ml | Standard Deviation 45 |
| Begining of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-8 (pg/ml) | 5785 pg/ml | Standard Deviation 859 |
| End of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | Neutrophil elastase activity (pg/ml) | 53 pg/ml | Standard Deviation 45 |
| End of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-8 (pg/ml) | 3428 pg/ml | Standard Deviation 2604 |
| End of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-17 (pg/ml) | 147 pg/ml | Standard Deviation 152 |
| End of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-1b (pg/ml) | 757 pg/ml | Standard Deviation 782 |
| End of Exacerbation | Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, and Neutrophil Elastase | IL-6 (pg/ml) | 37 pg/ml | Standard Deviation 17 |
Secondary
Sputum Inflammatory Mediators: IL-1β, IL-6, IL-8, IL-17A, TGF-β, TNF-α, and Neutrophil Elastase
In the Clinically Stable Cohort--Measurement of sputum inflammatory mediators: IL-1β, IL-6, IL-8, IL-17A, TGF-β, TNF-α, and neutrophil elastase
Time frame: Samples will be obtained at one outpatient clinic visit during the next calendar year
Population: Data not collected and will never be analyzed.