TY - JOUR
T1 - Progenitor cell-derived basophils
T2 - A novel barcoded passive degranulation assay in allergic diseases
AU - Wu, Jiakai
AU - Bahri, Rajia
AU - Tsoumani, Marina
AU - Semic-Jusufagic, Aida
AU - Murray, Clare S
AU - Custovic, Adnan
AU - Guibas, George V.
AU - Bennett, Miriam
AU - Wang, Ran
AU - Gauvreau, Gail
AU - Cusack, Ruth
AU - Mills, Clare
AU - Bulfone-Paus, Silvia
AU - Simpson, Angela
N1 - Funding Information:
We would like to thank NIHR Manchester Biomedical Research Centre, MRC Confidence in Concept 2015 – University of Manchester (MC_PC_15038) and JP Moulton Charitable Foundation, and the North West Lung Centre Charity, Medical Research Council (MRC) G0601361 (2007–2012), MR/K002449/1 (2013–2014) and MR/L012693/1 (2014–2018) for providing funding support of this work. We would also like to thank Manchester Allergy, Respiratory & Thoracic Surgery Biobank (ManARTS biobank) for providing the patient samples and clinical data. The clinical challenges were funded through the NIHR Clinical Research facility based at Manchester University NHS Foundation Trust and the European Union's Seventh Framework Program for research, technological development, and demonstration under grant agreement no. 312147 (Integrated Approaches to Food Allergen and Allergy Risk Management [iFAAM]). We also thank the Manchester Asthma and Allergy Study for providing the peanut tolerant samples and clinical data. Finally, we would like to thank Dr Gareth Howell for his invaluable help in flow cytometry technique, Dr Andrew Walls for supplying us with the BB1 antibody. Angela Simpson and Clare S Murray are supported by the Manchester Biomedical Research Centre.
Publisher Copyright:
© 2022 The Authors. Clinical & Experimental Allergy published by John Wiley & Sons Ltd.
PY - 2022/11/16
Y1 - 2022/11/16
N2 - Background: Effector cells assays provide an overall measure of responsiveness to allergen, but the lack of reliable and high-throughput assays limits the clinical utility. We aimed to develop a high-throughput basophil activation test based on human progenitor cell-derived basophils (PCB) and investigate the role of PCB activation test (PCBAT) in allergic diseases. Methods: Progenitor cell-derived basophils were differentiated from CD34+ progenitor cells and sensitized with sera from subjects sensitized to cat, peanut or atopic controls. Sensitized PCBs were stimulated with increasing concentrations of the corresponding allergens in vitro. Degranulation was assessed by measuring CD63 expression using flow cytometry. The correlations between PCBAT and clinical allergy were assessed. Results: Following passive sensitization of the mature PCBs with serum and allergen stimulation, an allergen specific dose-dependent increase in CD63 expression was observed. Sera from subjects sensitized to cat (n = 35, of which 17 subjects had clinical reactivity quantified using inhaled allergen challenge), peanut allergic (n = 30, of which 15 subjects had clinical reactivity validated using double blind, placebo controlled food challenges [DBPCFC]), peanut-sensitized but tolerant subjects (n = 5) were used to sensitize PCBs. PCBAT area under the curve (AUC) correlated with sIgE (r2 =.49, p =.001) in subjects sensitized to cat (sIgE ≥ 0.35KU/L). The provocation concentration of inhaled cat allergen (PC20) correlated with PCBAT AUC (r2 =.33, p =.016). In subjects sensitized to peanut, PCBAT AUC was highly correlated with sIgE to Ara h 2 (r2 =.59, p <.0001). Peanut threshold cumulative dose during DBPCFC was negatively correlated with PCBAT AUC (r2 =.57, p =.001) and IgE to Ara h1 (r2 =.55, p =.007), but not with sIgE to whole peanut or Ara h2. All peanut-sensitized but tolerant subjects showed no reaction to peanut on PCBAT. Conclusion: Progenitor cell-derived basophils activation test is a high-throughput assay, which correlates with clinical allergy and may confer a powerful alternative tool in allergy testing.
AB - Background: Effector cells assays provide an overall measure of responsiveness to allergen, but the lack of reliable and high-throughput assays limits the clinical utility. We aimed to develop a high-throughput basophil activation test based on human progenitor cell-derived basophils (PCB) and investigate the role of PCB activation test (PCBAT) in allergic diseases. Methods: Progenitor cell-derived basophils were differentiated from CD34+ progenitor cells and sensitized with sera from subjects sensitized to cat, peanut or atopic controls. Sensitized PCBs were stimulated with increasing concentrations of the corresponding allergens in vitro. Degranulation was assessed by measuring CD63 expression using flow cytometry. The correlations between PCBAT and clinical allergy were assessed. Results: Following passive sensitization of the mature PCBs with serum and allergen stimulation, an allergen specific dose-dependent increase in CD63 expression was observed. Sera from subjects sensitized to cat (n = 35, of which 17 subjects had clinical reactivity quantified using inhaled allergen challenge), peanut allergic (n = 30, of which 15 subjects had clinical reactivity validated using double blind, placebo controlled food challenges [DBPCFC]), peanut-sensitized but tolerant subjects (n = 5) were used to sensitize PCBs. PCBAT area under the curve (AUC) correlated with sIgE (r2 =.49, p =.001) in subjects sensitized to cat (sIgE ≥ 0.35KU/L). The provocation concentration of inhaled cat allergen (PC20) correlated with PCBAT AUC (r2 =.33, p =.016). In subjects sensitized to peanut, PCBAT AUC was highly correlated with sIgE to Ara h 2 (r2 =.59, p <.0001). Peanut threshold cumulative dose during DBPCFC was negatively correlated with PCBAT AUC (r2 =.57, p =.001) and IgE to Ara h1 (r2 =.55, p =.007), but not with sIgE to whole peanut or Ara h2. All peanut-sensitized but tolerant subjects showed no reaction to peanut on PCBAT. Conclusion: Progenitor cell-derived basophils activation test is a high-throughput assay, which correlates with clinical allergy and may confer a powerful alternative tool in allergy testing.
KW - allergy diagnosis
KW - asthma
KW - basophil
KW - cat allergy
KW - challenge tests
KW - flow cytometry
KW - peanut allergy
U2 - 10.1111/cea.14251
DO - 10.1111/cea.14251
M3 - Article
C2 - 36385515
AN - SCOPUS:85143316496
JO - Clinical and Experimental Allergy
JF - Clinical and Experimental Allergy
SN - 0954-7894
ER -
