SFR KO mice germinal center B cell RNA sequencing
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE147990 | GEO | 2020/11/16
REPOSITORIES: GEO
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Project description:Microarrays of gene expression in mouse germinal center light zone and dark zone B cells sorted according to the expression of cell surface molecules CD83 and CXCR4 We used microarray data to identify genes differentially expressed by B cells in the light and dark zones of germinal centers from mouse skin-draining lymph nodes 12 days after subcutaneous immunization with NP-OVA in alum. Germinal center B cells (B220+CD38-CD95+) were sorted from immunized mouse lymph nodes into light zone (CXCR4hi/CD83lo) and dark zone (CXCR4lo/CD83hi) populations, from which RNA was extracted
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Project description:A range of cellular interactions and cytokines influence immunoglobulin class switch recombination (CSR), which occurs both in extrafollicular antibody responses and germinal centers. This study addresses where CSR occurs during extrafollicular responses, allowing identification of the cellular interactions and cytokine-producing cells that influence CSR in vivo. The response in mice of B cells specific for 4-hydroxy-3-nitrophenyl-acetyl (NP) to NP-Ficoll was analyzed. Immunohistology and flow cytometry confirm responding NP-binding cells move to the outer T zone of the spleen and start dividing there on the second day. During the third day they move either to follicles as germinal center founding cells, or extrafollicular foci as plasmablasts. Analysis of expression of AID, Bcl-6 and Blimp-1 mRNA in single NP-binding cells indicates a proportion of T zone B blasts express AID transiently without Bcl-6 and these cells are shown to undergo CSR. By contrast Blimp-1+ cells emerging on the third day and were universally AID- and hence unable to initiate CSR. Bcl-6 mRNA without protein, expressed by 40% of naïve NP-binding cells, is rapidly downregulated. Bcl-6 and AID are coexpressed by germinal centre founding cells only on moving to follicles during the third day after immunization. Keywords: TaqMan Low Density Array
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