SARS-CoV-2 Spike Expression at the Surface of Infected Primary Human Airway Epithelial Cells
Shilei Ding, Damien Adam, Guillaume Beaudoin-Bussières, Alexandra Tauzin, Shang Yu Gong, Romain Gasser, Annemarie Laumaea, Sai Priya Anand, Anik Privé, Catherine Bourassa, Halima Medjahed, Jérémie Prévost, Hugues Charest, Jonathan Richard, Emmanuelle Brochiero , and Andrés Finzi
Centre de Recherche du CHUM (CRCHUM), Montréal
Different serological assays were rapidly generated to study humoral responses against the SARS-CoV-2 Spike glycoprotein. Due to the intrinsic difficulty of working with SARS-CoV-2 authentic virus, most serological assays use recombinant forms of the Spike glycoprotein or its receptor binding domain (RBD). Cell-based assays expressing different forms of the Spike, as well as pseudoviral assays, are also widely used. To evaluate whether these assays recapitulate findings generated when the Spike is expressed in its physiological context (at the surface of the infected primary cells), we developed an intracellular staining against the SARS-CoV-2 nucleocapsid (N) to distinguish infected from uninfected cells. Human airway epithelial cells (pAECs) were infected with authentic SARS-CoV-2 D614G or Alpha variants. We observed robust cell-surface expression of the SARS-CoV-2 Spike at the surface of the infected pAECs using the conformational-independent anti-S2 CV3-25 antibody. The infected cells were also readily recognized by plasma from convalescent and vaccinated individuals and correlated with several serological assays. This suggests that the antigenicity of the Spike present at the surface of the infected primary cells is maintained in serological assays involving the expression of the native full-length Spike.
Product use
Isolation and culture of primary human airway epithelial cells (pAECs)
Tissue type
Tissue info
Primary human airway epithelial cells (pAECs)

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