Antigen specificity and cross-reactivity drive functionally diverse anti–Aspergillus fumigatus T cell responses in cystic fibrosis
Carsten Schwarz, … , Alexander Scheffold, Petra Bacher
Carsten Schwarz, … , Alexander Scheffold, Petra Bacher
Published January 26, 2023
Citation Information: J Clin Invest. 2023;133(5):e161593. https://doi.org/10.1172/JCI161593.
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Clinical Research and Public Health Immunology Pulmonology

Antigen specificity and cross-reactivity drive functionally diverse anti–Aspergillus fumigatus T cell responses in cystic fibrosis

Abstract

BACKGROUND The fungus Aspergillus fumigatus causes a variety of clinical phenotypes in patients with cystic fibrosis (pwCF). Th cells orchestrate immune responses against fungi, but the types of A. fumigatus–specific Th cells in pwCF and their contribution to protective immunity or inflammation remain poorly characterized.METHODS We used antigen-reactive T cell enrichment (ARTE) to investigate fungus-reactive Th cells in peripheral blood of pwCF and healthy controls.RESULTS We show that clonally expanded, high-avidity A. fumigatus–specific effector Th cells, which were absent in healthy donors, developed in pwCF. Individual patients were characterized by distinct Th1-, Th2-, or Th17-dominated responses that remained stable over several years. These different Th subsets target different A. fumigatus proteins, indicating that differential antigen uptake and presentation directs Th cell subset development. Patients with allergic bronchopulmonary aspergillosis (ABPA) are characterized by high frequencies of Th2 cells that cross-recognize various filamentous fungi.CONCLUSION Our data highlight the development of heterogenous Th responses targeting different protein fractions of a single fungal pathogen and identify the development of multispecies cross-reactive Th2 cells as a potential risk factor for ABPA.FUNDING German Research Foundation (DFG), under Germany’s Excellence Strategy (EXC 2167-390884018 “Precision Medicine in Chronic Inflammation” and EXC 2051-390713860 “Balance of the Microverse”); Oskar Helene Heim Stiftung; Christiane Herzog Stiftung; Mukoviszidose Institut gGmb; German Cystic Fibrosis Association Mukoviszidose e.V; German Federal Ministry of Education and Science (BMBF) InfectControl 2020 Projects AnDiPath (BMBF 03ZZ0838A+B).

Authors

Carsten Schwarz, Patience Eschenhagen, Henrijette Schmidt, Thordis Hohnstein, Christina Iwert, Claudia Grehn, Jobst Roehmel, Eva Steinke, Mirjam Stahl, Laura Lozza, Ekaterina Tikhonova, Elisa Rosati, Ulrik Stervbo, Nina Babel, Jochen G. Mainz, Hilmar Wisplinghoff, Frank Ebel, Lei-Jie Jia, Matthew G. Blango, Peter Hortschansky, Sascha Brunke, Bernhard Hube, Axel A. Brakhage, Olaf Kniemeyer, Alexander Scheffold, Petra Bacher

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Figure 4

A. fumigatus–reactive Th cell subsets recognize different protein targets.

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A. fumigatus–reactive Th cell subsets recognize different protein targe...
(A) PBMCs from pwCF (n = 8–14) were stimulated with whole A. fumigatus lysate, and reactive CD154+ Tmem, Th1 (IFN-γ+), Th17 (IL-17A+), or Th2 (CRTH2+) cells were FACS sorted, expanded, and restimulated with a panel of single A. fumigatus proteins. Reactivity is indicated as the percentage of CD154+TNF-α+ within CD4+ T cells. (B) Th2 target proteins (Aspf2, Aspf3, CpcB, CatB, Fg-gap, GliT) and non-Th2 target proteins (Scw4, Aspf22, Pst1, Shm2, CcpA, TpiA, Crf1, Sod3) were pooled and used for ex vivo stimulation of PBMCs from A. fumigatus–sensitized pwCF. Relative cytokine production within reactive CD154+ Tmem cells (upper plots) and absolute frequencies of cytokine producers within CD4+ T cells (lower plots) are shown (IL-4, IFN-γ, IL-17A, n = 20; IL-5, IL-13, n = 13). (C) A. fumigatus–reactive T cell lines were generated as in A and restimulated with different A. fumigatus antigen extracts in the presence of autologous FastDCs derived from blood monocytes. Reactivity in relation to restimulation with the initially used total A. fumigatus lysate is shown. (D) Reactivity of expanded CD154+ Tmem cells from Th2-high (n = 7) or Th2-low (n = 6) patients to the different A. fumigatus protein fractions. Each symbol in A–D represents 1 donor; horizontal lines indicate the mean in upper plots and the geometric mean in lower plots of B; truncated violin plots with the quartiles and range are shown in C and D. Statistical differences were determined by 2-tailed, paired Wilcoxon rank test in B. and by Kruskal-Wallis test with Dunn’s post hoc test in C and D.