Divergent SARS CoV-2 Omicron-reactive T- and B cell responses in COVID-19 vaccine recipients

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Divergent SARS CoV-2 Omicron-reactive T- and B cell responses in COVID-19 vaccine recipients
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Vaccines produce robust Tcell immunity against SARSCoV2 variants. Neutralizing antibodies were lowest against Omicron but can be partially restored with booster vaccination. SciImmunology

Serum was collected in 10-ml tubes without anticoagulant, centrifuged at 2500 rpm for 15 min, aliquoted, and stored at −20°C for further experiments. PBMCs were isolated from blood collected in KEDTA or Lithium Heparin tubes by density gradient centrifugation. Briefly, blood was layered on a density gradient , and PBMCs were separated by centrifuging at 2000 rpm for 30 min.

Viral genome sequences were determined using Illumina deep-sequencing. RNA was extracted using AMPure XP beads and cDNA was generated using ProtoscriptII reverse transcriptase enzyme according to the manufacturer’s protocol . Samples were amplified using the QIAseq SARS-CoV-2 Primer Panel . Amplicons were purified with 0.

The beta variant contained the following spike changes: L18F, D80A, D215G, del241-243, K417N, E484K, N501Y, D614G, and A701V. The delta variant contained the following spike changes: T19R, G142D, del156-157, R158G, A222V, L452R, T478K, D614G, P681R and D950N.

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