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Comparison of gene expression in NOD versus B6 splenic B cell subsets.

ABSTRACT: NOD mice are an inbred strain that display enhanced MZ B cell differentiation from an early age. Interestingly, several lines of evidence implicate MZ B cells in this strain as important contributors to the T cell mediated beta cell destruction associated with the development of type 1 diabetes (T1D). In order to develop a better understanding of the underlying causes for augmented MZ B cell production in NOD mice, we obtained the transcriptional profiles of FO and MZ subsets and TR precursors from NOD mice and compared them to those of the B6 strain. Red cell depleted splenocyte suspensions from from eight pooled NOD or B6 mice were stained with biotinylated anti-CD3Îµ (145-2C11) and CD11b (M1/70, BD Biosciences) antibodies followed by anti-biotin Microbeads (Miltenyi Biotec, Bergisch Gladbach, Germany). Separation of unlabeled B cells from samples was achieved to a >93% purity (as determined by flow cytometry) by negative depletion with an autoMACS cell sorter (Miltenyi Biotec). These cells were then stained with a cocktail of: non-stimulatory Affinipure goat anti-mouse-IgM-FITC Fab fragments from Jackson Immunoresearch; anti-CD21/35-PE (7G6), anti-CD24-biotin (30-F1; with streptavidin-PerCP secondary), B220-APC-Cy7 (RA36B2) from BD Biosciences; anti-CD93-APC (AA4.1), anti-CD23-PE-Cy7 (B3B4) from eBiosciences. T1, T2FO, T3, FO, T2MZ and MZ subsets were then separated with a FACSAria Cell Sorter (BD Biosciences). This process was repeated in an independent experiment in order to generate a duplicate sample of each B cell subset from NOD and B6 mice. Labelled cRNA from B cell samples were hybridized to Mouse Genome 430 2.0 Arrays.

ORGANISM(S): Mus musculus

SUBMITTER: Pablo Silveira

PROVIDER: E-GEOD-43396 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Comparison of gene expression in NOD versus B6 splenic B cell subsets.

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