Induced Tolerogenic Dendritic Cells as Modulators of Allergic Asthma (MGH-002)
Despite advances in medications, allergic diseases, including allergic asthma continue to rise in prevalence. For this reason, there is a need for a better understanding of the mechanisms of allergic diseases and novel insights into modulating allergic inflammation. The investigators hypothesize that much remains to be learned about the behavior of T effector and T regulatory cells in allergic disease. Furthermore, the investigators hypothesize that novel mechanisms of allergic tolerance may exist, and elucidation of these mechanisms may provide insights into novel therapeutic strategies to control allergic diseases. The investigators will investigate the capacity for T cell tolerance induction in allergic subjects by a novel type of immune tolerizing dendritic cell (it-DC). The investigators will assess whether in vitro generated it-DCs have the capacity to induce antigen-specific T regulatory cells and suppress allergen-specific T effector cell function in vitro.
Standardized Cat Allergen extract and Dust Mite Allergens will be used to generate changes in the airways that occur during exposure to allergen. For this investigation, the route of administration will be topical application of the titrated allergen to a bronchoscopically isolated subsegment of one lobe of one lung. The dose of biologic will be determined from prior skin-prick testing.
|Asthma Allergies||Biological: Bronchoscopy, Segmental Allergen Challenge and Bronchoalveolar Lavage Procedure: Phlebotomy||Phase 1|
|Study Design:||Intervention Model: Single Group Assignment
Masking: Open Label
Primary Purpose: Basic Science
|Official Title:||Induced Tolerogenic Dendritic Cells as Modulators of Allergic Asthma|
- Effect of it-DC on the phenotype, function, proliferation and survival of allergen-specific effector T cells. [ Time Frame: 24 hours ]
We will generate induced tolerogenic dendritic cells (it-DCs) in vitro from peripheral blood monocytes and dendritic cells isolated from each allergic asthmatic subject and assess their capacity to (a) attenuate effector CD4+ T cell (Teff) responses and/or (b) induce T regulatory (Treg) cells from Teff cells as well as CD4+ naïve and memory T cells. Attenuation of T cell responses (a) will be assessed by proliferation assays and cytokine production. Moreover, in order to evaluate the functionality of the it-DC induced Tregs (b), co-culture suppression assays using cat or house dust-mite allergen-specific Teff cells isolated from BAL following segmental allergen challenge (SAC) as responder cells, will be examined. The percent suppression of responding Teff cells will be assessed by examining Teff cell proliferation and survival/apoptosis, and by cytokine production.
(Note: SAC samples from healthy controls are not involved in the primary analysis.)
- Percent distribution of peripheral blood DC subsets [ Time Frame: 4 weeks ]We will determine the frequency of each peripheral blood DC subset from allergic asthmatic subjects at 0 and 4 weeks using cell surface staining/flow cytometry analysis. Results will be expressed in terms of percent distribution of each DC subset within the total DC pool.
- Percent suppression of naïve T cells (Tn) and memory T cells (Tmem) proliferation by it-DC-induced Tregs from each subject. [ Time Frame: 4 weeks ]The ability of it-DC-induced Tregs from each allergic asthmatic to suppress proliferation of autologous CD4+ Tn and Tmem cells in response to anti-CD3 will be assessed and expressed as a percent suppression of proliferation using an in vitro proliferation assay.
- Differences in proliferation by allergen-specific Teff cells from BAL and blood. [ Time Frame: 4 weeks ]We will measure the proliferative responses in it-DC co-culture assays by allergen-specific CD4+ Teff cells obtained from the BAL of allergic asthmatic subjects as compared to those of blood-derived CD4+ Tmem cells from the same subjects and from 10 healthy controls.
|Study Start Date:||January 2013|
|Study Completion Date:||September 2016|
|Primary Completion Date:||September 2016 (Final data collection date for primary outcome measure)|
Experimental: Allergic Asthmatic, Non-allergic non-asthmatics(HC)
Track 1: Adult subjects who are allergic asthmatics or non-allergic non-asthmatics(Healthy Controls) will not receive segmental allergen challenge to the lung but will have their blood drawn at 1 time point. Track 2: Adult subjects who are allergic asthmatics will receive segmental allergen challenge to the lung and have their blood drawn at 2 time points.
Biological: Bronchoscopy, Segmental Allergen Challenge and Bronchoalveolar Lavage
On the first bronchoscopy day,BAL will first be performed in the lingual without instillation of diluent or allergen. A 2ml aliquot of isotonic diluent is instilled into the right upper lobe.The procedure is repeated in the right middle lobe with instillation of 2ml of standardized cat or mite allergen solution. A test dose concentration of allergen is administered first consisting of 2 ml of allergen at 1/10th(Cat,D.farinae)or 1/30th(D.pteronyssinus)the threshold concentration.If on visual inspection through the bronchoscope there is no evidence of mucosal inflammation after 2 minutes a second segmental allergen challenge will be done in the right middle lobe using 2ml of full-dose allergen at the threshold concentration(Cat,D.farinae)or 1/3 the threshold concentration(D.pteronyssinus). This dose will be predetermined by quantitative skin prick testing.A second bronchoscopy is performed 24 hours after delivery of allergen extract and diluent to obtain BAL fluid from the RUL and RML.
Other Names:Procedure: Phlebotomy
200 (40 teaspoons) ml of blood will be obtained from the subjects to collect different leukocyte populations. A second 200 ml of blood will be obtained from the Track 2 subjects 4 weeks later for isolation of T lymphocytes for functional studies.
Other Name: Blood Draw
This is a single-center, open-label, controlled mechanistic study before-and-after allergen instillation study that uses each subject as his/her own control. Reliability of study measurements of expression is accomplished by comparing repeated measurements. Additional controls are provided by measuring the expression of markers known to have stable expression, including cell structural proteins and chemokine markers that the investigators know are induced in this experimental paradigm from our previous studies. This is an investigational study. Subjects will receive an investigational product, either Standardized Cat Allergen Extract or Standardized Dust Mite Allergen (Dermatophagoides farinae or D. pteronyssinus). No subject will be given a placebo. There are 2 cohorts, allergic asthmatics (AA) and healthy controls (HC). The main comparisons will be measurement of primary and secondary outcome measures in: (1) diluent versus allergen-challenged segments of the lung in each asthmatic subject and (2) comparison of these responses in allergic asthmatics (AA) and healthy controls (HC).
The investigators will enroll 20 AA subjects (10 in Track 1 & 10 in Track 2) and 10 HC subjects (Track 1). There will be no randomization. The choice of allergen will be determined by allergy skin testing. When a subject is allergic to both cat and dust mite, one will be selected with a goal of having a representative sample of cat and dust mite challenges in each cohort. The 10 allergic asthmatic (AA) subjects and HC subjects in Track 1 will undergo preliminary screening tests and provide 200 ml of blood for DC and T cell studies on one occasion. Track 1 subjects will not undergo bronchoscopy, bronchoalveolar lavage, or segmental allergen challenge. The 10 allergic asthmatic (AA) subjects in Track 2 will undergo SAC with either standardized cat or mite allergen. These subjects will also donate 200 ml of blood on two occasions separated by a minimum of 4 weeks and not more than 12 weeks.
Please refer to this study by its ClinicalTrials.gov identifier: NCT01711593
|United States, Massachusetts|
|Massachusetts General Hospital|
|Boston, Massachusetts, United States, 02114|
|Principal Investigator:||Andrew D Luster, M.D., Ph.D.||Massachusetts General Hospital|