Immunohistochemistry for protein detection in PFA-fixed paraffin-embedded SARS-CoV-2-infected COPD airway epithelium

STAR Protoc. 2021 Sep 17;2(3):100663. doi: 10.1016/j.xpro.2021.100663. Epub 2021 Jun 18.

Abstract

Patients with chronic lung disease are vulnerable to getting severe diseases associated with SARS-CoV-2 infection. Here, we describe protocols for subculturing and differentiating primary normal human bronchial epithelial (NHBE) cells of patients with chronic obstructive lung disease. The differentiation of NHBE cells in air-liquid interface mimics an in vivo airway and provides an in vitro model for studying SARS-CoV-2 infection. We also describe a protocol for detecting proteins in the sectioned epithelium for detailing SARS-CoV-2 infection-induced pathobiology with a vertical view.

Keywords: Antibody; Health Sciences; Immunology; Microbiology; Microscopy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Bronchi / metabolism*
  • Bronchi / pathology
  • Bronchi / virology
  • COVID-19 / complications*
  • COVID-19 / metabolism
  • COVID-19 / virology
  • Coronavirus Nucleocapsid Proteins / analysis*
  • Coronavirus Nucleocapsid Proteins / metabolism
  • Epithelium / metabolism*
  • Epithelium / pathology
  • Epithelium / virology
  • Humans
  • Immunohistochemistry
  • Paraffin Embedding
  • Phosphoproteins / analysis
  • Phosphoproteins / metabolism
  • Pulmonary Disease, Chronic Obstructive / metabolism
  • Pulmonary Disease, Chronic Obstructive / pathology*
  • Pulmonary Disease, Chronic Obstructive / virology
  • SARS-CoV-2 / isolation & purification*
  • Virus Replication

Substances

  • Coronavirus Nucleocapsid Proteins
  • Phosphoproteins
  • nucleocapsid phosphoprotein, SARS-CoV-2