Comparison of gene expression in NOD versus NOD.NOR-Chr4 (NR4) splenic B cells.

ABSTRACT: Type 1 Diabetes (T1D) in humans and the non-obese diabetic (NOD) mouse model results from autoreactive T cell destruction of pancreatic beta cells. A pathogenic role for B lymphocytes (B cells) in T1D first became evident when NOD mice made deficient in this population through introduction of an inactivated IgM-BM-5 heavy chain gene (NOD.IgM-BM-5null) or chronic treatment with anti-IgM antibodies were strongly protected from disease. We produced an NOD background strain developing a greatly decreased T1D incidence due to a NOR-derived 44.31Mb congenic region from rs3674285 to D4Mit127 on distal Chr. 4 (termed NOD.NOR-Chr4 (NR4)) containing disease resistance alleles decreasing the pathogenic activity of autoreactive B cells. Microarrays were conducted on B cells purified from spleens of NOD and NR4 mice to highlight differentially expressed genes within the distal Chr. 4 locus. B cells were either cultured in media alone (unstimulated) or with BCR cross-linking anti-IgM-F(abM-bM-^@M-^Y)2 fragments (stimulated) for 2h before RNA was extracted for transcript analysis. B cells from three independent lots of pooled spleens (n=7-8) from NOD or NR4 female mice, respectively, were purified using a magnetic-activated cell sorting (MACS) negative depletion strategy. Purified B cells from each lot were resuspended at 1x10<7> cells/ml for 2h in complete RPMI media alone or with 10M-BM-5g/ml AffiniPure goat anti-mouse IgM F(abM-bM-^@M-^Y)2 fragments. RNA was then purified from B cells and subjected to one-cycle linear amplification and biotin labeling. Two BCR-stimulated and three unstimulated NOD B cell samples plus three BCR-stimulated and three unstimulated NR4 B cell samples were hybridized to Mouse Genome 430 2.0 Arrays.

ORGANISM(S): Mus musculus

SUBMITTER: Pablo Silveira 

PROVIDER: E-GEOD-37294 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress