Ignite Creation Date:
2024-05-06 @ 1:44 PM
Last Modification Date:
2024-10-26 @ 1:18 PM
Study NCT ID:
NCT04100902
Status:
COMPLETED
Last Update Posted:
2022-05-10
First Post:
2019-09-23
Brief Title:
The Effect of Allergen Immunotherapy on Anti-viral Immunity in Patients With Allergic Asthma
Sponsor:
Bispebjerg Hospital
Organization:
Bispebjerg Hospital
Study Overview
Official Title:
The Effect of Allergen Immunotherapy on Anti-viral Immunity in Patients With Allergic Asthma - A Randomized Double-blind Placebo-controlled Trial of HDM-AIT
Status:
COMPLETED
Status Verified Date:
2022-05
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:
VITAL
Brief Summary:
Aim To investigate the possible immune modulatory effects of allergen immunotherapy AIT on respiratory immunity in patients with allergic asthma AA
Background Allergic sensitization to aeroallergens is a common co-morbidity in asthma that is associated with more frequent and severe asthma attacks The investigators have recently shown that patients with allergic asthma also have an increased risk of pneumonia and hence allergy in asthma may be associated with a relative respiratory immunodeficiency However the increased risk was obliterated in patients treated with AIT
Methods Patients with asthma sensitized to house-dust mite HDM is enrolled in a randomized double-blind placebo-controlled study of HDM-AIT Patients will be scheduled for 9 visits through 8 months including randomization to 6 months of treatment with either HDM-AIT AcarizaxOdactra or placebo Primary interferons IFN type I and III will be investigated in human bronchial epithelial cells as the primary outcome Secondary outcomes such as Inflammatory cytokines immunologic phenotype and immunohistochemistry will be investigated in bronchial biopsies blood bronchoalveolar lavage fluid sputum and HDM-patch biopsies as well as a thorough respiratory and allergic evaluation
Expected outcomes The investigators expect that patients with AA have 1 decreased production of anti-viral type I and III IFN and that AIT increases these measures 2 Anti-bacterial response is reduced through IL12 ß-defensin and IFN-γ and that AIT increases these measures 3 Lastly the investigators expect that T-cell response is dysregulated Th11Th2 in patients with AA and that these findings are modulated in an immuno-protective direction after AIT
Perspectives This project will expand our understanding of the clinical significance of allergy in asthma in a completely novel direction and show how AIT may modulate the immune response to prevent infections
Background Allergic sensitization to aeroallergens is a common co-morbidity in asthma that is associated with more frequent and severe asthma attacks The investigators have recently shown that patients with allergic asthma also have an increased risk of pneumonia and hence allergy in asthma may be associated with a relative respiratory immunodeficiency However the increased risk was obliterated in patients treated with AIT
Methods Patients with asthma sensitized to house-dust mite HDM is enrolled in a randomized double-blind placebo-controlled study of HDM-AIT Patients will be scheduled for 9 visits through 8 months including randomization to 6 months of treatment with either HDM-AIT AcarizaxOdactra or placebo Primary interferons IFN type I and III will be investigated in human bronchial epithelial cells as the primary outcome Secondary outcomes such as Inflammatory cytokines immunologic phenotype and immunohistochemistry will be investigated in bronchial biopsies blood bronchoalveolar lavage fluid sputum and HDM-patch biopsies as well as a thorough respiratory and allergic evaluation
Expected outcomes The investigators expect that patients with AA have 1 decreased production of anti-viral type I and III IFN and that AIT increases these measures 2 Anti-bacterial response is reduced through IL12 ß-defensin and IFN-γ and that AIT increases these measures 3 Lastly the investigators expect that T-cell response is dysregulated Th11Th2 in patients with AA and that these findings are modulated in an immuno-protective direction after AIT
Perspectives This project will expand our understanding of the clinical significance of allergy in asthma in a completely novel direction and show how AIT may modulate the immune response to prevent infections
Detailed Description:
11 Background Respiratory viral infections are the major cause of acute worsening of asthma Importantly too these infections may also predispose to bacterial infections Patients with allergic asthma suffer from more frequent lower respiratory tract infections more severe and longer lasting lower respiratory tract symptoms compared to healthy controls Hence the need to use antibiotics may reflect in part the increased susceptibility to viral infection in allergic asthma The investigators have recently demonstrated that patients with allergic asthma have an increased risk of being prescribed antibiotics for respiratory infections compared to non-allergic patients with asthma Other studies have identified allergic sensitization and respiratory viral infections as synergetic risk factors in asthma exacerbations These findings suggest a possible link between the allergic sensitization asthma and an impaired immune response against respiratory infections Furthermore the investigators have found that allergen-specific immunotherapy AIT reduces the risk of being prescribed antibiotics for respiratory infections in patients with allergic asthma These novel observations make it of interest to investigate mechanistic pathways in target cells subjected to AIT
In a recent study investigating potential mechanisms involved in the interaction between allergen and viral infection the investigators found that house dust mite HDM impairs viral stimulus TLR3-induced type I and type III interferons in bronchial epithelial cells from asthmatic patients and similarly in an in vivo mouse model of asthma exacerbation This novel finding was accompanied with a direct effect of HDM on reducing TLR3 glycosylation Hence HDM exposure in patients with allergic asthma may also contribute to a defect antiviral response by interfering with components important for the anti-viral signalling such as TLR3 Furthermore the investigators have observed that HDM has a capacity to induce over-expression of upstream Th2 cytokines such as IL-33 which may contribute to asthma exacerbations In our studies HDM stands out amongst different allergens in causing both release of inflammatory inducing damage-associated metabolic patterns and impaired interferon response
Inferentially in this study the investigators will have the unique opportunity to examine effects of human in vivo HDM-AIT on the bronchial epithelial cell production of type I and type III interferons as well as the balance between Th1and Th2 inflammation in response to viral and bacterial triggers The investigators speculate that exposure to HDM-allergen may predispose to asthma exacerbations by causing dysregulation of the anti-viral interferon system as well as Th1and Th2 inflammation and then that desensitization of patients with HDM-sensitive allergic asthma AA increases innate interferon response
12 Hypothesis
1 Viral induced type I and type III IFN is increased in HBECS from patients with allergic asthma sensitized to HDM after 24 weeks of Acarizax
2 Viral induced type I and III IFN response is impaired in HBECs from patients with allergic asthma sensitized to HDM before 24 weeks of Acarizax
13 Investigational Medical Product IMP ACARIZAX Acarizax is sublingual allergen immunotherapy SLIT for the treatment of HDM allergic rhinitis AR and allergic asthma AA Acarizax is composed of standardized allergen extract from the HDMs Dermatophagoides pteronyssinus and Dermatophagoides farina
The efficacy of Acarizax has been tested in randomized double-blinded placebo-controlled trials as add-on treatment for the best maintenance treatment for AR and AA The clinical effect of Acarizax has been demonstrated 8-14 weeks after initiation of treatment and data on clinical effect is available for up to 18 months of treatment Acarizax was found to reduce time to first exacerbation hazard-ratio 07 in patients with AA Acarizax has been shown to reduce daily mean inhaled corticosteroid budesonide by 81 microgramday 95 CI 27-136microgramday In AR Acarizax has been shown to reduce Total Combined Rhinitis Score TCRS A meta-analysis on the efficacy of AIT on atopic dermatitis shows moderate level of efficacy Hence we will be studying the potential change in the immunologic phenotype as an exploratory outcome
AIT has been known for more than 100 years as a potential curative and specific treatment for allergic diseases The mechanism by which AIT works is poorly understood but a key target is likely modulation of immunopathology Dysregulation of the immune system is thought to play an essential role in atopic diseases and is influenced by multiple complex regulatory mechanisms Well-established immune modulatory effects of AIT are classified into following type of events
Table 1 Mechanisms of AIT Order of events Events Change Time to initial change
1 Basophil- and Mast cell activity and degranulation of IgE mediated histamine release Decrease Immediately
2 Induction of allergen-specific Treg- and Breg cells and suppression of allergen specific Th1 and Th2 cells Increase Days to weeks
3 Specific IgE Early increase followed by chronic decrease to baseline Weeks to years
4 Specific IgG4 Continuously increase throughout treatment duration Weeks
5 Tissue resident eosinophils basophils and mast cells and release of their mediators Decrease Months
6 Type 1 skin reactivity decrease Months
14 Rationale
Anti-viral immunity Interferons IFNs are essential cytokines in the anti-viral defence IFNs are produced by many cell types in the lungs where human bronchial epithelial cells HBECs and plasmacytoid dendritic cells pDCs are the main producers The major effects of IFNs include blocking of viral invasion into neighbour cells by cleaving viral nucleic acid and inhibiting viral replication and inducing apoptosis in affected cells HBECs are critical for the development of a sufficient innate immune response to viral infection Increasing evidence suggest that HBECS from patients with asthma and allergic asthma exhibit an impaired ability to secrete type I IFN-beta and III IFN-lambda interferons at viral infection In addition IFN-beta and IFN-lambda may inversely correlate with the level of Type 2 inflammation in the airway epithelium
pDCs are essential effectors of the primary innate immune defence against viral infections releasing large amounts of IFN-alpha and driving Th1 polarisation of CD4 lymphocytes However pDCs also play an important role as antigen-presenting cells by which they may drive a Th2 polarisation increasing IgE-production in B-cells and potential cross-linking of FcepsilonR1-alpha receptors Interestingly it has recently been demonstrated that viral induced IFN-alpha release was diminished by activation of the FcepsilonR1-alpha receptor on the pDC by allergen exposure suggesting a potential link between allergy and a reduced viral defence In patients with allergic asthma viral induced IFN-alpha release was impaired in PBMCs indicating a more systemic impact in response to bronchial infection
AIT Mechanisms leading to suppression of allergic inflammation In general it is acknowledged that Th2 Treg cells tolerance is decisive for the development or suppression of allergic inflammation Treg cells secrete high amounts of essential suppressor cytokines IL-10 and TGF-beta These cytokines contribute to the control of Th2 mediated allergic disease in various ways Treg cells regulates B-cells by inducing a competitive IgG4 and IgA response resulting in suppression of IgE Furthermore Treg cells may suppress activation and degranulation of mast cells basophils and eosinophils Lastly Treg cells suppress Th2 cells homing to tissue and epithelial cell activation and production of proinflammatory cytokines Breg cells control excessive inflammatory responses through secretion of IL-10 supports differentiation of Treg cells and induces synthesis of IgG4 These effects may be involved in response to AIT and correlate with clinical improvement Thus AIT may result in a decrease of bronchial nasal and conjunctival hyperreactivity which is a reflection of decrease in underlying mucosal inflammation Anti-IgE treatment ameliorates the seasonal variations in asthma exacerbations associated with virus infections which may be related to an increase in the initial interferon response to viral infections We thus speculate that the potential immunomodulatory effects of AIT resulting in lowered risk of prescription antibiotics is the net result of alteration in allergen-induced mucosal Th-2 inflammation mediated by an increase in primary IFN defence responses to viral infections
66 Study governance and oversight
661 Study Site Bispebjerg Hospital Dep of Respiratory Medicine Bispebjerg Bakke 23 Entrance 66 DK - 2400 Copenhagen NV Denmark Phone 45 35 31 35 69 Fax 45 35 31 21 79
6612 Additional sites of analyses
Department of Immunology and Microbiology University of Copenhagen
Department of Respiratory Immunopharmacology University of Lund
ALK-Abelló AS Boege Allé 6-8 2970 Hoersholm
6613 Study registration and authorities Study code VITAL EudraCT 2019-003261-18 Ethics Committee journal number H-19052148 Danish Medicines Agency 2019-003261-18 ClinicalTrialgov NCT04100902
6614 Steering Committee Christian Uggerhoej Woehlk MD PhD-student Asger Sverrild MD PhD Charlotte Menné Bonefeld Professor Lena Uller Ass Professor Steen Roenborg MD PhD Celeste Porsbjerg Professor MD
6615 Participants and collaborators
1 Pharmacy Region Hovedstadens Apotek Marielundvej 25 2730 Herlev Denmark
2 Peter Arvidsson MD PhD ALK-Abelló Nordic AS Kungbacke Sweden
3 Peter Sejer Andersen ALK-Abelló AS Boege Allé 6-8 Bygn 9 2970 Hoersholm
4 Peter Adler Würtzen Senior Specialist PhD ALK-Abelló AS Boege Allé 6-8 Bygn 9 2970 Hoersholm
8 STATISTICAL ANALYSES 81 Sample size estimate Sample size calculations have been carried out using G-Power software Behaviour Research Methods 2007 Faul et al
811 Primary outcome
To our knowledge there is no preceding studies investigation the relationship between AIT treatment and change in IFN-response in HBECS The investigators have previously shown that treating human airway epithelium in vitro with azithromycin increase the IFN-response to viral infection mimics in a dose-dependent manner Fold-change in INF-β was 155 SD 0555 at the given concentration Given a significant change in IFN-β from week 0 to week 24 is set to be a 155-fold change with SD of 0555 that alpha is set to 005 and the power to 080 in a two-sided test design the calculation would look like this
Mean fold change in INF-β group 1 placebo 1 Mean fold change in INF-β group 2 Acarizax 155 SD 055 16 subjects will be needed in each study arm Assuming a drop-out of 20 a minimum of 40 subjects in total will need to be randomized
82 Definitions of analysis sets 1 Populations for analysis
1 Intention-to-treat population All randomized subjects who receive at least one dose of Acarizax or placebo A comparison of the treatment groups will be performed
2 Population for analysis based on adherence 80 to treatment
1 Intention-to-treat population All randomized subjects with adherence 80 to Acarizax therapy or placebo A comparison of the treatment groups will be performed
821 Efficacy analysis set Intention-to-treat population as defined in 82
822 Safety analysis set All patients who receive at least one dose of Acarizaxplacebo
83 Methods for statistical analyses All data limited to primary and key secondary outcomes will be analysed using SPSS v 14 or later all data are reported as median values with interquartile ranges and were analysed using non-parametric tests Data within each group Acarizax and placebo will be analysed using linear mixed model or ANCOVA Statistical differences between the Acarizax and placebo groups will be determined using the Mann - Whitney U test for unrelated samples Skewed-data will be log-transformed before analysis and reported using median and 2575 percentiles Parametric data will be analysed using the above-mentioned methods Categorial data will be analysed using χ²-test Statistical significance is defined as p005 The data from the subject dropping out will still be used in the analysis using methods for handling of missing data
831 Analysis of the primary variables The fold-change in IFN-β and IFN-λ are analysed before and after RV infection andor polyIC stimulation from V3 to V12 and will be compared between intervention and placebo treatment The investigators expect parametric distribution of data
The calculation will be as follows
1 deltaIFN-β IFN-β V12 - IFN-βV3
2 Mean fold-change IFN-β Sum of individual changes N
Supportive primary objectives are calculated between intervention and placebo treatment groups
1 Change in expression of IFN-λ measured in ρgml ELISA and reported as fold change mean SD
2 Change in viral loadTCID50 assay measure reported as fold change mean SD
832 Analysis of the secondary variables
Changes in IFN-responses will be analysed using same procedure as analysis of the primary outcome
Cytokines will be analysed accordingly to the method described in 831 and reported as ρmml or similar quantitative method and reported as fold change mean SEM
Change in IFN-βTSLP ratio and reported in
The change expressed as a ratio in number of airway submucosal muscle and epithelium mast cells eosinophils neutrophils NK-cells MCT MCTC and MCCPA3 per mm2 determined by microscopic evaluation of mucosa biopsies and epithelial brushings from V2 to V7
833 Interim analysis No interim analysis is planned
834 Use of database REDCap will be used for online CRF data management
9 STUDY AND DATAMANGEMENT 91 Economy
The scientific ideas of protocol are based solely on the work of members of the steering committee None of the members of the steering committee have any commercial interest in the results of this study As per 010819 the project has received
1 Unrestricted grant from ALK-Abelló Nordic of 900000dkk
2 Unrestricted grant from Sawmill owner Jeppe Juhl and wife Ovita Juhls memorial fund 250000 dkk
3 HARBOEFONDEN DKK 200000
4 Pharmaxis Osmohale Mannitol Challenge tests
92 Subject reimbursement Each subject completing the study will receive a total of 6 months of Acarizax treatment equivalent to approximate 5500 dkk Subject randomized to Acarizax arm will not be furtherly compensated
921 Source data Will be filed at Bispebjerg Hospital Denmark study site in accordance with The Danish Data Protection Agency laws Source data will be kept for a maximum of 20 years from 01092019 At this point all source data will be destroyed
922 Study agreements All agreements relating to the study will be available in the trial master file TMF
923 Archiving of study documents Documents will be archived at Bispebjerg Hospital Denmark study site in accordance with GCP and The Danish Data Protection Agency laws
93 Monitoring of the study
The study will be monitored by the GCP Unit in Copenhagen
Copenhagen University Hospital GCP-unit Bispebjerg Hospital Building 51 3fl Bispebjerg Bakke 23 DK - 2400 København NV Denmark Phone 45 3531 3887 E-mail gcp-enhedenbispebjerg-frederiksberg-hospitalerregionhdk wwwgcp-enheddk
In a recent study investigating potential mechanisms involved in the interaction between allergen and viral infection the investigators found that house dust mite HDM impairs viral stimulus TLR3-induced type I and type III interferons in bronchial epithelial cells from asthmatic patients and similarly in an in vivo mouse model of asthma exacerbation This novel finding was accompanied with a direct effect of HDM on reducing TLR3 glycosylation Hence HDM exposure in patients with allergic asthma may also contribute to a defect antiviral response by interfering with components important for the anti-viral signalling such as TLR3 Furthermore the investigators have observed that HDM has a capacity to induce over-expression of upstream Th2 cytokines such as IL-33 which may contribute to asthma exacerbations In our studies HDM stands out amongst different allergens in causing both release of inflammatory inducing damage-associated metabolic patterns and impaired interferon response
Inferentially in this study the investigators will have the unique opportunity to examine effects of human in vivo HDM-AIT on the bronchial epithelial cell production of type I and type III interferons as well as the balance between Th1and Th2 inflammation in response to viral and bacterial triggers The investigators speculate that exposure to HDM-allergen may predispose to asthma exacerbations by causing dysregulation of the anti-viral interferon system as well as Th1and Th2 inflammation and then that desensitization of patients with HDM-sensitive allergic asthma AA increases innate interferon response
12 Hypothesis
1 Viral induced type I and type III IFN is increased in HBECS from patients with allergic asthma sensitized to HDM after 24 weeks of Acarizax
2 Viral induced type I and III IFN response is impaired in HBECs from patients with allergic asthma sensitized to HDM before 24 weeks of Acarizax
13 Investigational Medical Product IMP ACARIZAX Acarizax is sublingual allergen immunotherapy SLIT for the treatment of HDM allergic rhinitis AR and allergic asthma AA Acarizax is composed of standardized allergen extract from the HDMs Dermatophagoides pteronyssinus and Dermatophagoides farina
The efficacy of Acarizax has been tested in randomized double-blinded placebo-controlled trials as add-on treatment for the best maintenance treatment for AR and AA The clinical effect of Acarizax has been demonstrated 8-14 weeks after initiation of treatment and data on clinical effect is available for up to 18 months of treatment Acarizax was found to reduce time to first exacerbation hazard-ratio 07 in patients with AA Acarizax has been shown to reduce daily mean inhaled corticosteroid budesonide by 81 microgramday 95 CI 27-136microgramday In AR Acarizax has been shown to reduce Total Combined Rhinitis Score TCRS A meta-analysis on the efficacy of AIT on atopic dermatitis shows moderate level of efficacy Hence we will be studying the potential change in the immunologic phenotype as an exploratory outcome
AIT has been known for more than 100 years as a potential curative and specific treatment for allergic diseases The mechanism by which AIT works is poorly understood but a key target is likely modulation of immunopathology Dysregulation of the immune system is thought to play an essential role in atopic diseases and is influenced by multiple complex regulatory mechanisms Well-established immune modulatory effects of AIT are classified into following type of events
Table 1 Mechanisms of AIT Order of events Events Change Time to initial change
1 Basophil- and Mast cell activity and degranulation of IgE mediated histamine release Decrease Immediately
2 Induction of allergen-specific Treg- and Breg cells and suppression of allergen specific Th1 and Th2 cells Increase Days to weeks
3 Specific IgE Early increase followed by chronic decrease to baseline Weeks to years
4 Specific IgG4 Continuously increase throughout treatment duration Weeks
5 Tissue resident eosinophils basophils and mast cells and release of their mediators Decrease Months
6 Type 1 skin reactivity decrease Months
14 Rationale
Anti-viral immunity Interferons IFNs are essential cytokines in the anti-viral defence IFNs are produced by many cell types in the lungs where human bronchial epithelial cells HBECs and plasmacytoid dendritic cells pDCs are the main producers The major effects of IFNs include blocking of viral invasion into neighbour cells by cleaving viral nucleic acid and inhibiting viral replication and inducing apoptosis in affected cells HBECs are critical for the development of a sufficient innate immune response to viral infection Increasing evidence suggest that HBECS from patients with asthma and allergic asthma exhibit an impaired ability to secrete type I IFN-beta and III IFN-lambda interferons at viral infection In addition IFN-beta and IFN-lambda may inversely correlate with the level of Type 2 inflammation in the airway epithelium
pDCs are essential effectors of the primary innate immune defence against viral infections releasing large amounts of IFN-alpha and driving Th1 polarisation of CD4 lymphocytes However pDCs also play an important role as antigen-presenting cells by which they may drive a Th2 polarisation increasing IgE-production in B-cells and potential cross-linking of FcepsilonR1-alpha receptors Interestingly it has recently been demonstrated that viral induced IFN-alpha release was diminished by activation of the FcepsilonR1-alpha receptor on the pDC by allergen exposure suggesting a potential link between allergy and a reduced viral defence In patients with allergic asthma viral induced IFN-alpha release was impaired in PBMCs indicating a more systemic impact in response to bronchial infection
AIT Mechanisms leading to suppression of allergic inflammation In general it is acknowledged that Th2 Treg cells tolerance is decisive for the development or suppression of allergic inflammation Treg cells secrete high amounts of essential suppressor cytokines IL-10 and TGF-beta These cytokines contribute to the control of Th2 mediated allergic disease in various ways Treg cells regulates B-cells by inducing a competitive IgG4 and IgA response resulting in suppression of IgE Furthermore Treg cells may suppress activation and degranulation of mast cells basophils and eosinophils Lastly Treg cells suppress Th2 cells homing to tissue and epithelial cell activation and production of proinflammatory cytokines Breg cells control excessive inflammatory responses through secretion of IL-10 supports differentiation of Treg cells and induces synthesis of IgG4 These effects may be involved in response to AIT and correlate with clinical improvement Thus AIT may result in a decrease of bronchial nasal and conjunctival hyperreactivity which is a reflection of decrease in underlying mucosal inflammation Anti-IgE treatment ameliorates the seasonal variations in asthma exacerbations associated with virus infections which may be related to an increase in the initial interferon response to viral infections We thus speculate that the potential immunomodulatory effects of AIT resulting in lowered risk of prescription antibiotics is the net result of alteration in allergen-induced mucosal Th-2 inflammation mediated by an increase in primary IFN defence responses to viral infections
66 Study governance and oversight
661 Study Site Bispebjerg Hospital Dep of Respiratory Medicine Bispebjerg Bakke 23 Entrance 66 DK - 2400 Copenhagen NV Denmark Phone 45 35 31 35 69 Fax 45 35 31 21 79
6612 Additional sites of analyses
Department of Immunology and Microbiology University of Copenhagen
Department of Respiratory Immunopharmacology University of Lund
ALK-Abelló AS Boege Allé 6-8 2970 Hoersholm
6613 Study registration and authorities Study code VITAL EudraCT 2019-003261-18 Ethics Committee journal number H-19052148 Danish Medicines Agency 2019-003261-18 ClinicalTrialgov NCT04100902
6614 Steering Committee Christian Uggerhoej Woehlk MD PhD-student Asger Sverrild MD PhD Charlotte Menné Bonefeld Professor Lena Uller Ass Professor Steen Roenborg MD PhD Celeste Porsbjerg Professor MD
6615 Participants and collaborators
1 Pharmacy Region Hovedstadens Apotek Marielundvej 25 2730 Herlev Denmark
2 Peter Arvidsson MD PhD ALK-Abelló Nordic AS Kungbacke Sweden
3 Peter Sejer Andersen ALK-Abelló AS Boege Allé 6-8 Bygn 9 2970 Hoersholm
4 Peter Adler Würtzen Senior Specialist PhD ALK-Abelló AS Boege Allé 6-8 Bygn 9 2970 Hoersholm
8 STATISTICAL ANALYSES 81 Sample size estimate Sample size calculations have been carried out using G-Power software Behaviour Research Methods 2007 Faul et al
811 Primary outcome
To our knowledge there is no preceding studies investigation the relationship between AIT treatment and change in IFN-response in HBECS The investigators have previously shown that treating human airway epithelium in vitro with azithromycin increase the IFN-response to viral infection mimics in a dose-dependent manner Fold-change in INF-β was 155 SD 0555 at the given concentration Given a significant change in IFN-β from week 0 to week 24 is set to be a 155-fold change with SD of 0555 that alpha is set to 005 and the power to 080 in a two-sided test design the calculation would look like this
Mean fold change in INF-β group 1 placebo 1 Mean fold change in INF-β group 2 Acarizax 155 SD 055 16 subjects will be needed in each study arm Assuming a drop-out of 20 a minimum of 40 subjects in total will need to be randomized
82 Definitions of analysis sets 1 Populations for analysis
1 Intention-to-treat population All randomized subjects who receive at least one dose of Acarizax or placebo A comparison of the treatment groups will be performed
2 Population for analysis based on adherence 80 to treatment
1 Intention-to-treat population All randomized subjects with adherence 80 to Acarizax therapy or placebo A comparison of the treatment groups will be performed
821 Efficacy analysis set Intention-to-treat population as defined in 82
822 Safety analysis set All patients who receive at least one dose of Acarizaxplacebo
83 Methods for statistical analyses All data limited to primary and key secondary outcomes will be analysed using SPSS v 14 or later all data are reported as median values with interquartile ranges and were analysed using non-parametric tests Data within each group Acarizax and placebo will be analysed using linear mixed model or ANCOVA Statistical differences between the Acarizax and placebo groups will be determined using the Mann - Whitney U test for unrelated samples Skewed-data will be log-transformed before analysis and reported using median and 2575 percentiles Parametric data will be analysed using the above-mentioned methods Categorial data will be analysed using χ²-test Statistical significance is defined as p005 The data from the subject dropping out will still be used in the analysis using methods for handling of missing data
831 Analysis of the primary variables The fold-change in IFN-β and IFN-λ are analysed before and after RV infection andor polyIC stimulation from V3 to V12 and will be compared between intervention and placebo treatment The investigators expect parametric distribution of data
The calculation will be as follows
1 deltaIFN-β IFN-β V12 - IFN-βV3
2 Mean fold-change IFN-β Sum of individual changes N
Supportive primary objectives are calculated between intervention and placebo treatment groups
1 Change in expression of IFN-λ measured in ρgml ELISA and reported as fold change mean SD
2 Change in viral loadTCID50 assay measure reported as fold change mean SD
832 Analysis of the secondary variables
Changes in IFN-responses will be analysed using same procedure as analysis of the primary outcome
Cytokines will be analysed accordingly to the method described in 831 and reported as ρmml or similar quantitative method and reported as fold change mean SEM
Change in IFN-βTSLP ratio and reported in
The change expressed as a ratio in number of airway submucosal muscle and epithelium mast cells eosinophils neutrophils NK-cells MCT MCTC and MCCPA3 per mm2 determined by microscopic evaluation of mucosa biopsies and epithelial brushings from V2 to V7
833 Interim analysis No interim analysis is planned
834 Use of database REDCap will be used for online CRF data management
9 STUDY AND DATAMANGEMENT 91 Economy
The scientific ideas of protocol are based solely on the work of members of the steering committee None of the members of the steering committee have any commercial interest in the results of this study As per 010819 the project has received
1 Unrestricted grant from ALK-Abelló Nordic of 900000dkk
2 Unrestricted grant from Sawmill owner Jeppe Juhl and wife Ovita Juhls memorial fund 250000 dkk
3 HARBOEFONDEN DKK 200000
4 Pharmaxis Osmohale Mannitol Challenge tests
92 Subject reimbursement Each subject completing the study will receive a total of 6 months of Acarizax treatment equivalent to approximate 5500 dkk Subject randomized to Acarizax arm will not be furtherly compensated
921 Source data Will be filed at Bispebjerg Hospital Denmark study site in accordance with The Danish Data Protection Agency laws Source data will be kept for a maximum of 20 years from 01092019 At this point all source data will be destroyed
922 Study agreements All agreements relating to the study will be available in the trial master file TMF
923 Archiving of study documents Documents will be archived at Bispebjerg Hospital Denmark study site in accordance with GCP and The Danish Data Protection Agency laws
93 Monitoring of the study
The study will be monitored by the GCP Unit in Copenhagen
Copenhagen University Hospital GCP-unit Bispebjerg Hospital Building 51 3fl Bispebjerg Bakke 23 DK - 2400 København NV Denmark Phone 45 3531 3887 E-mail gcp-enhedenbispebjerg-frederiksberg-hospitalerregionhdk wwwgcp-enheddk
Study Oversight
Has Oversight DMC:
None
Is a FDA Regulated Drug?:
True
Is a FDA Regulated Device?:
False
Is an Unapproved Device?:
None
Is a PPSD?:
None
Is a US Export?:
True
Is an FDA AA801 Violation?:
None