Recycling of memory B cell supports affinity maturation to antigenic drift

This study aims to compare relative mRNA transcript levels between murine naïve B cells, germinal centre B cells, emigrating memory B cells in draining lymph node and circulating memory B cells in distal lymph nodes. Overall design: Four B cell subsets were isolated by fluorescence activated cell sorting and processed for RNA sequencing. Mice used were C57BL/6J background. Germinal centre and emigrating memory B cells were isolated from mGFP-expressing cells in popliteal lymph nodes among IgHNP Cg1cre k-/- mTmG adoptive cell transfer recipients immunised with 4-hydroxy-3-nitrophenylacetyl (NP) conjugated to chicken gamma globulin (CGG) in alum 8 days prior to isolation of cells. Circulating memory B cells were isolated from the axillary and brachial lymph nodes of the same mice at d8 post-immunisation with NP-CGG. Naive B cells were isolated from the non-adoptively transferred cells demarcated as those expressing neither mGFP or mTomato in axillary and brachial lymph nodes.