Nature research paper: Long-primed germinal centres with enduring affinity maturation and clonal migration
For the MD39 plus alum bolus group, RMs were housed at the Tulane National Primate Research Center as part of a larger NHP study . This study was approved by the Tulane University IACUC. Animals were grouped together to match age, weight and gender. Animals were aged 3.5–5.0 years at the time of first immunization, with three females and three males in the study group.
For LN FNA data inclusion in GC gating, a threshold of 250 B cells in the sample was used and, for Env-binding GC B cell gating, a threshold of 75 GC B cells was used. Any sample with fewer than 75 GC B cells but with a B cell count of more than 500 cells was set to a baseline of 0.001% EnvGC B cells ; otherwise, the limit of detection was calculated based on the median of ) from pre-immunization LN FNA samples.
The following reagents were used for staining: Alexa Fluor 647 streptavidin , BV421 streptavidin , BV711 streptavidin , PE streptavidin , Live/Dead fixable aqua , Propidium iodide , eBioscience Fixable Viability Dye eFluor 780 , mouse anti-human CD20 BV785, BUV395, Alexa Fluor 488, PerCP-Cy5.5 , mouse anti-human IgM PerCP-Cy5.
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