Contribution of cysteine and serine proteases to proteolytic digestion in an allergy-eliciting house dust mite

• Published in: Journal of insect physiology Vol. 133; p. 104285
• Main Authors: Vidal-Quist, José Cristian, Ortego, Félix, Hernández-Crespo, Pedro
• Format: Journal Article
• Language: English
• Published: Elsevier Ltd 01.08.2021
• Subjects: Allergens; Coprophagy; Guanine; Immunotherapy; Protease inhibitors; Zymogen activation; HDM; Allergens; Zymogen activation; Protease inhibitors; Immunotherapy; Coprophagy; WMC; CP; SP; Guanine
• ISSN: 0022-1910; 1879-1611
• DOI: 10.1016/j.jinsphys.2021.104285

[Display omitted] •Body-associated proteolytic activity in HDM is dependent on cysteine proteases.•Major allergen Der p 1 accounts for most of the cysteine protease activity.•Fecal-associated proteolytic activity depends on cysteine and serine proteases.•Ingested cystatin A, by inhibiting Der p 1, impairs activation of serine proteases.•Feeding experiments demonstrate the autocoprophagic behavior of the mite. The digestive physiology of house dust mites (HDM) is of interest to understand their allergenicity towards humans since many of their allergens are digestive enzymes and/or are excreted into airborne fecal pellets. The aim of this study is to provide insight on the biochemical basis of proteolytic digestion in Dermatophagoides pteronyssinus, the most widespread HDM species. First, assays using non-specific protein substrates on purified fecal and body extracts determined that body-associated activity is almost exclusively dependent on cysteine proteases, and specifically on major allergen Der p 1. By contrast, cysteine and serine proteases contributed similarly to the activity estimated on fecal extracts. Second, the screening of group-specific peptide-based protease inhibitors followed by ingestion bioassays revealed that the human skin-derived cysteine protease inhibitor cystatin A produces a significant reduction in mite feeding (i.e. excreted guanine), and triggers the overproduction of Der p 1 (3-fold increase by ELISA). Noteworthy, the inhibition of cysteine proteases by cystatin A also resulted in a reduction in three non-target serine protease activities. Further incubation of these extracts with exogenous Der p 1, but not with other commercial cysteine proteases, restored trypsin (Der p 3) and chymotrypsin (Der p 6) activities, indicating that Der p 1 is responsible for their activation in vivo. Finally, the role of serine proteases on the mite's digestive physiology is discussed based on their remarkable activity in fecal extracts and the autocoprophagic behavior reported in mites in this study.