Neutrophilic inflammation is a hallmark of some specific asthma phenotypes; its etiology is not fully understood yet. House dust mite (HDM) is the most common factor involving with the pathogenesis of airway inflammation. This study aims to elucidate the role of cross-antibodies against HDM-derived factors in the development of neutrophilic inflammation in the airway.Blood samples were collected from asthma patients with chronic neutrophilic asthma to be analysed the HDM-specific cross-reactive antibodies. The role of an antibody against HDM-derived enolase (EnoAb) in the impairment of airway epithelial barrier function and induction of airway inflammation was assessed in a cell culture model and an animal model.High similarity (72%) of the enolase gene sequences was identified between HDM and human. Serum EnoAb was detected in patients with chronic neutrophilic asthma. The EnoAb bound to airway epithelial cells to form complexes with enolase, which activated complements, impaired airway epithelial barrier functions and induced neutrophilic inflammation in the airway tissues.HDM-derived enolase can induce specific cross-antibodies by human, which induce neutrophilic inflammation in the airway.