Identification and characterization of immunogenic rna species in hdm allergens that modulate eosinophilic lung inflammation

Hamad H. Alanazi, Li She, Xiao Dong Li

Research output: Contribution to journalArticlepeer-review

Abstract

Environmental allergens such as house dust mites (HDM) are often in complex forms containing both allergic proteins that drive aberrant type 2 responses and microbial substances that induce innate immune responses. These allergen-associated microbial components play an important role in regulating the development of type 2 inflammatory conditions such as allergic asthma. However, the underlying mechanisms remain largely undefined. The protocol presented here determines the structural characteristics and in vivo activity of allergen-associated immunostimulatory RNA. Specifically, common allergens are examined for the presence of double-stranded RNA (dsRNA) species that can stimulate IFN responses in lungs and restrain the development of severe lung eosinophilia in a mouse model of HDM-induced allergic asthma. Here, we have included the following three assays: Dot blot to show the dsRNA structures in total RNA isolated from allergens including HDM species, RT-qPCR to measure the activities of HDM RNA in interferon stimulating genes (ISGs) expression in mouse lungs and FACS analysis to determine the effects of HDM RNA on the number of eosinophils in BAL and lung, respectively.

Original languageEnglish (US)
Article numbere61183
Pages (from-to)1-10
Number of pages10
JournalJournal of Visualized Experiments
Volume2020
Issue number159
DOIs
StatePublished - May 2020

Keywords

  • BAL
  • Bronchoalveolar lavage
  • DsRNA
  • FACS
  • Fluorescence-activated cell sorting
  • HDM
  • House dust mites
  • ISGs
  • Immunology and Infection
  • Interferon stimulated genes
  • Issue 159
  • PAMP
  • Pathogen-associated molecular pattern
  • RT-qPCR

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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