Project description:We examined the transcriptional changes that occur as Antibody Secreting Cells (ASC) differentiate and mature using an in vitro culture system. ASC were profiled directly post isolation and at time points weekly out to 48 days in culture. These data reveal how the long-lived ASC transcriptome matures over time.
Project description:We examined the chromatin accessibility changes that occur as antibody-secreting cells (ASC) differentiate and mature using an in vitro culture system. ASC were profiled directly post isolation and at time points weekly out to 14 days in culture. These data reveal how the long-lived ASC accessible chromatin matures over time.
Project description:We measured genes expression differences in antibody secreting cells from LynKO mice in following treatment with a BMI-1 inhibitor (PTC-028) or a vehicle control. BMI-1 inhibition lead to a reduction in antibody secreting cells in LynKO mice and in humans donors. We performed RNA sequencing to understand the impact of BMI-1 inhibition on antibody secreting cells in LynKO mice.
Project description:The CD19 positive antibody secreting cells (ASC) in both bone marrow (BM) have the capacity to provide immune memory in addition to cells traditionally considered long-lived, the CD19-negative BM ASC. We performed flow cytometry (FCM) immunophenotyping, fluorescence-activated cell sorting (FACS) for cell subset isolation, ELISpot assays detecting the isotype of antibody secretion as well as antibodies against vaccine derived antigens, and comparative gene expression analyses of CD19- ASC, CD19+ ASC, CD20- B cells, and CD20+ B cells from BM. The findings may aid in the understanding of the differential cell subsets created through vaccination and lead to improved vaccine strategies and production. FACS sorted tissue B cells and antibody secreting cell subset gene expression.
Project description:We report whole transcriptome RNASeq data for cell-sorted pop2, pop3, and pop5 which are antibody-secreting cells from human peripheral blood
Project description:Following activation by cognate antigen, B cells undergo fine-tuning of their antigen receptors and may ultimately differentiate into antibody-secreting cells (ASCs). While antigen-specific antibodies from B cell receptor (BCR) expressing B cells can be readily cloned and sequenced following flow sorting, antigen-specific plasma cells that lack surface BCR cannot be easily profiled in a high-throughput way. Here, we report an approach, TRAPnSeq (antigen specificity mapping through Ig secretionTRAPandSequencing), that allows capture of secreted antibodies on the surface of ASCs, which in turn enables high-throughput screening of single ASCs against large antigen panels and recovery of paired VH:VL antibody sequences. This approach incorporates flow cytometry, standard microfluidic platforms and DNA barcoding technologies to isolate and characterize antigen-specific ASCs through single cell V(D)J, RNA and antigen barcode sequencing. We show the utility of TRAPnSeq by profiling antigen-specific IgG and IgE plasma cells from mouse and humans and validate antigen binding by ELISA. TRAPnSeq can easily be combined with existing B cell platforms to accelerate antibody discovery from ASCs and can further be expanded to any protein secreting-cells.
Project description:The differentiation of activated B cells into antibody secreting plasma cells (PCs) is governed by a strict regulatory network that results in expression of specific transcriptomes along the continuum. In vitro models resulting in PCs that are phenotypically identical to the in vivo state permit investigation of molecules, transcripts, pathways, and metabolomes not previously identified in the differentiation process. We used an in vitro system and RNA-seq to identify protein-coding and non-coding RNA (ncRNA) transcripts expressed in human resting and activated B cells, and PCs. Analysis of immunoglobulin (Ig) transcripts showed that the repertoire was not skewed, and that memory B cells preferentially expanded and differentiated into PCs. Of interest, we identified more than 980 differentially expressed ncRNA transcripts displaying stage-specific patterns of increased or decreased expression across differentiation some of which are known to target transcription, proliferation, cytoskeletal, autophagy and proteasome pathways. Interestingly, ncRNAs located within Ig loci may be targeting both Ig and non-Ig related transcripts. ncRNAs associated with B cell malignancies were also identified in this model. Taken together, this system provides a platform to study the detailed role of specific ncRNAs in B cell differentiation and altered expression of those ncRNAs involved in B cell malignancies.