Mice can make neutralizing antibodies to SARSCoV2 via a complementary pathway not requiring follicular helper T cells or GerminalCenters, says new research from SciImmunology.
mice were infected with 70 PFU of PR8. All work with SARS-CoV-2 was performed in a biosafety level 3 facility with approval from the office of Environmental Health and Safety and the Institutional Animal Care and Use Committee at Yale University.Used vials of Moderna mRNA-1273 and Pfizer-BioNTech BNT162b2 mRNA vaccine were obtained from Yale Health within 6 hours of opening. All vials contained less than one full dose per vial, and no vaccines were diverted for the purpose of this study.
For cell sorting, medLN were stained with aforementioned B cell surface markers and viability dye, along with 2 μg/ml FLAG-tagged SARS-CoV-2 spike protein for 30 min at 4°C . Cells were then stained with both APC- and PE-conjugated anti-FLAG antibodies to double-stain spike-specific B cells. Spike-specific plasmablasts were sorted into 350 μl Buffer RLT Plus with 1% β-mercaptoethanol. RLT lysate was vortexed for 1 min, frozen on dry ice, and then stored at -80°C.
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