Single-cell RNA sequencing identifies precise tolerogenic cellular and molecular pathways induced by depigmented-polymerized grass pollen allergen extract - 04/05/23
Graphical abstract |
Abstract |
Background |
Immunologic mechanism of action of allergoids remains poorly understood. Previous models of allergenicity and immunogenicity have yielded suboptimal knowledge of these immunotherapeutic vaccine products. Novel single-cell RNA sequencing technology offers a bridge to this gap in knowledge.
Objective |
We sought to identify the underpinning tolerogenic molecular and cellular mechanisms of depigmented-polymerized Phleum pratense (Phl p) extract.
Methods |
The molecular mechanisms underlying native Phl p, depigmented Phl p (DPG–Phl p), and depigmented-polymerized (DPG-POL–Phl p) allergoid were investigated by single-cell RNA sequencing. Allergen-specific TH2A, T follicular helper (Tfh), and IL-10+ regulatory B cells were quantified by flow cytometry in peripheral blood mononuclear cells from 16 grass pollen–allergic and 8 nonatopic control subjects. The ability of Phl p, DPG–Phl p, and DPG-POL–Phl p to elicit FcεRI- and FcεRII-mediated IgE responses was measured by basophil activation test and IgE-facilitated allergen binding assay.
Results |
Analysis revealed that DPG-POL–Phl p downregulated genes associated with TH2 signaling, induced functional regulatory T cells exhibiting immunosuppressive roles through CD52 and Siglec-10, modulated genes encoding immunoproteasome that dysregulate the processing and presentation of antigens to T cells and promoted a shift from IgE toward an IgA1 and IgG responses. In grass pollen–allergic subjects, DPG-POL–Phl p exhibited reduced capacity to elicit proliferation of TH2A, IL-4+ Tfh and IL-21+ Tfh cells while being the most prominent at inducing IL-10+CD19+CD5hi and IL-10+CD19+CD5hiCD38intCD24int regulatory B-cell subsets compared to Phl p (all P < .05). Furthermore, DPG-POL–Phl p demonstrated a hypoallergenic profile through basophil activation and histamine release compared to Phl p (31.54-fold, P < .001).
Conclusions |
Single-cell RNA sequencing provides an in-depth resolution of the mechanisms underlying the tolerogenic profile of DPG-POL–Phl p.
Le texte complet de cet article est disponible en PDF.Key words : Depigmented-polymerized allergen, allergen immunotherapy, allergic rhinitis, allergoid, mechanism of action, single-cell RNA-Seq
Abbreviations used : AIT, Breg, DAO, DPG–Phl p, DPG-POL–Phl p, GPA, IgE-FAB, MS, NAC, PBMC, Phl p, scRNA-Seq, Tfh, TH2A, Treg
Plan
| The first 2 authors contributed equally to this article, and both should be considered first author. The last 2 authors contributed equally to this article, and both should be considered senior author. |
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| This work was supported by LETI Pharma SL and the NIHR Imperial Biomedical Research Centre (BRC). The views expressed are those of the author(s) and not necessarily those of the NIHR or the Department of Health and Social Care. |
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| Disclosure of potential conflict of interest: J. A. Layhadi reports grants via Biomedical Research Funding (Imperial College BRC), all outside the submitted work. S. R. Durham reports grants from Imperial College Innovations during the conduct of the study; grants from Regeneron, USA, grants from Biotech Tools, grants from ALK, Denmark, personal fees from Anergis, Switzerland, personal fees from Adiga, USA, personal fees from Biomay, Austria, personal fees from Allergy Therapeutics, UK, personal fees from Med Update GmbH, Germany, grants from Food Standards Agency, UK, and grants from National Institute of Health Research, UK, all outside the submitted work. M. H. Shamji reports research grants from Immune Tolerance Network, Medical Research Council, Allergy Therapeutics, LETI Laboratorios, and Rovolo Biotherapeutics, and lecture fees from Allergy Therapeutics and LETI Laboratorios, all outside the submitted work. R. Moya and J. Carnés are employees of LETI Pharma SL. The rest of the authors declare that they have no relevant conflicts of interest. |
Vol 151 - N° 5
P. 1357 - mai 2023 Retour au numéro
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