Project description:Purpose: We aimed to link different variations of memory B cell subsets to their proliferating precursors via shared transcriptomic features. Methods : Spleens were processed according to the manuscript. Cells were collected via FACS, washed twice in PBS + 1% FCS, resuspended in 350 ul RLT Plus Buffer (Qiagen) with 1% 2-Me, and frozen at -80 ⁰C. Results: Memory cells have many DEGs in comparison to FO naïve cells. However, differences between memory cells themselves can be attributed to imprinting by their respective proliferating precursors. DP MBCs are split into two major subsets based on transcriptional state. A germinal center B cell program may be required to establish some DEGs in GC-derived memory cells.
Project description:Purpose: We aimed to link different variations of memory B cell subsets to their proliferating precursors via shared epigenetic features. Methods : Spleens were processed according to the manuscript. Cells were sorted and DNA was isolated using a protocol adapted from Corces et al. Samples were sequenced to obtain 20M 2x75 bp paired-end reads using an Illumina NextSeq 550 sequencer (Illumina, Inc, California, USA). Results: Memory cells have many DARs in comparison to FO naïve cells. However, differences between memory cells themselves can be attributed to imprinting by their respective proliferating precursors. A germinal center B cell program may be required to establish some DARs in GC-derived memory cells.
Project description:Purpose: We wanted to explore further heterogeneity of MBC subsets that were previously defined by our lab. Methods: MBC subsets were sorted and stained with unique anti-mouse HTO antibodies (CITE-seq). Results: We found that heterogeneity among MBC subsets was most prominent among DP MBC subsets, which could be separated by extrafollicular or germinal center transcriptional signatures, whereas DN and SP MBCs mostly exhibited an extrafollicular signature.