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Virus neutralization assays for human respiratory syncytial virus using airway organoids
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Virus neutralization assays for human respiratory syncytial virus using airway organoids
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Journal Article
Virus neutralization assays for human respiratory syncytial virus using airway organoids
2024
Overview
Neutralizing antibodies are considered a correlate of protection against severe human respiratory syncytial virus (HRSV) disease. Currently, HRSV neutralization assays are performed on immortalized cell lines like Vero or A549 cells. It is known that assays on these cell lines exclusively detect neutralizing antibodies (nAbs) directed to the fusion (F) protein. For the detection of nAbs directed to the glycoprotein (G), ciliated epithelial cells expressing the cellular receptor CX3CR1 are required, but generation of primary cell cultures is expensive and labor-intensive. Here, we developed a high-throughput neutralization assay based on the interaction between clinically relevant HRSV grown on primary cells with ciliated epithelial cells, and validated this assay using a panel of infant sera. To develop the high-throughput neutralization assay, we established a culture of differentiated apical-out airway organoids (Ap-O AO). CX3CR1 expression was confirmed, and both F- and G-specific monoclonal antibodies neutralized HRSV in the Ap-O AO. In a side-by-side neutralization assay on Vero cells and Ap-O AO, neutralizing antibody levels in sera from 125 infants correlated well, although titers on Ap-O AO were consistently lower. We speculate that these lower titers might be an actual reflection of the neutralizing antibody capacity in vivo. The organoid-based neutralization assay described here holds promise for further characterization of correlates of protection against HRSV disease.
Publisher
Springer International Publishing,Springer Nature B.V
Subject
/ Antibodies, Monoclonal - immunology
/ Antibodies, Neutralizing - immunology
/ Antibodies, Viral - immunology
/ Assaying
/ Biomedical and Life Sciences
/ CX3C Chemokine Receptor 1 - immunology
/ CX3C Chemokine Receptor 1 - metabolism
/ Epithelial Cells - immunology
/ Epithelial Cells - metabolism
/ Humans
/ Infant
/ Infants
/ Neutralization Tests - methods
/ Original
/ Respiratory Syncytial Virus Infections - immunology
/ Respiratory Syncytial Virus Infections - virology
/ Respiratory Syncytial Virus, Human - immunology
/ Viral Fusion Proteins - immunology
/ Viral Fusion Proteins - metabolism
/ Viruses
