For identification of GCs by flow cytometry, cells isolated from lymph nodes collected at the 5th (after HF or DMSO priming), 14th, 28th (after booster), 42nd, and 56th days were passed through a 40-μm cell strainer to generate single-cell suspensions, after which they were stained with Alexa Fluor 488–labeled anti-mouse B220 (eBioscience), Alexa Fluor 647–labeled anti-mouse total IgG (Cell Signaling Technology), or biotin-labeled anti-GL7 (eBioscience) for 1 hour at 4°C, followed by Alexa Fluor 555–labeled streptavidin (Invitrogen) for 1 hour at room temperature. The stained cells were fixed using 4% paraformaldehyde, washed with FACS buffer, and data were acquired on a BD LSRFortessa (BD Biosciences) cytometer and analyzed using FlowJo (Tree Star Inc.).

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