Epithelial Interleukin‐1 Receptor‐Like‐1 Activation Is Contingent on Interleukin‐33 Isoforms and Asthma‐Related Receptor Variation

ABSTRACT Introduction The interleukin‐33/interleukin‐1 receptor‐like‐1 (IL‐33/IL1RL1) signalling pathway is implicated in asthma pathogenesis, with IL1RL1 nonsynonymous genetic polymorphisms associated with disease risk. We aimed to determine these variants' effect on IL1RL1 signalling induced by different IL33 isoforms thought to be elevated in the asthmatic airway. Method In a project funded by GSK plc, which has developed an IL‐33 receptor inhibitor for asthma treatment, human embryonic kidney 293 (HEK293) cells expressing secreted embryonic alkaline phosphatase (SEAP) driven by a nuclear factor kappa‐beta (NF‐κB) promoter, were transiently transfected with IL1RL1 , containing one of four extracellular and Toll/interleukin 1 receptor (TIR) domain haplotypes. Cells were stimulated with seven different splice and proteolytic‐generated IL‐33 isoforms (0.001–50 ng/mL) for 24 h. Supernatant SEAP activity and interleukin‐8 (IL‐8) levels were determined. Primary human bronchial epithelial cells (HBECs) representing different genotype carriers were stimulated with IL‐33 112–270 (50 ng/mL) and induced IL‐8 mRNA expression measured. Results HEK293 cells carrying both asthma extracellular and TIR domain IL1RL1 risk haplotypes presented maximal IL33‐driven signalling, with minimal signalling after IL‐33 activation in other protective haplotypes. All IL‐33 isoforms activated IL1RL1 but with differing magnitudes. Proteolytically cleaved IL33 95–270 and IL33 106–270 had the greatest effect and the IL33 113–270 , and Exon 3,4 deletion isoform exhibited the lowest. The effect of extracellular and TIR domain genetic variants on receptor signalling was replicated in primary HBECs. Maximal IL1RL1 signalling was observed in cells carrying both extracellular and TIR signalling domain risk haplotypes. Conclusions Overall, our study suggests asthma patients carrying the extracellular and TIR domain risk haplotype and have a lung microenvironment that promotes elevated levels of cleaved IL33, particularly where IL33 95–270 and IL33 106–270 may be more amenable to IL33/IL1RL1 targeting.