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Transcriptomic profiling of purified polyclonal anti-dsDNA antibodies stimulated human mesangial cells

ABSTRACT: Analysis genes which alter after anti-dsDNA antibodies stimulation in human mesangial cells. The hypothesis is anti-dsDNA antibodies might affected a pro-inflammatory cytokine gene or genes in cell proliferation and apoptosis pathways

ORGANISM(S): Homo sapiens

PROVIDER: GSE80364 | GEO | 2016/10/01

SECONDARY ACCESSION(S): PRJNA318742

REPOSITORIES: GEO

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Homo sapiens

Project description:Transcriptomic profiling of purified polyclonal anti-dsDNA antibodies stimulated human mesangial cells

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Expression data from dsDNA-stimulated mouse embryonic fibroblasts

Project description:Transfection of dsDNA into many mammalian cell types indues the production of type I interferons and interferon-stimulated genes. We performed an siRNA screen to identify genes involved in this innate immune response, and identified Abcf1. We used microarrays to determine which genes are regulated by ABCF1 following dsDNA stimulation. We treated p53-/- mouse embryonic fibroblasts (MEFs) with siRNAs targeting Abcf1 or Irf3 (positive control) or a negative control siRNA (siCtrl), and then stimulated the cells with 45-base pair dsDNA for 6 hrs or left the cells unstimulated. We then lysed the cells and hybridized the RNA to Affymetrix microarrays.

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Expression data from dsDNA-stimulated mouse embryonic fibroblasts

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Multiple tolerance checkpoints restrain affinity maturation of B cells expressing the germline precursor of a lupus patient-derived anti-dsDNA antibody in knock-in mice

Project description:Anti-dsDNA antibodies are a hallmark of systemic lupus erythematosus and are highly associated with its exacerbation. Cumulative evidence has suggested that somatic hypermutation contributes to the high-affinity reactivity of anti-dsDNA antibodies. Our previous study demonstrated that these antibodies are generated from germline precursors with low-affinity ssDNA reactivity through affinity maturation and clonal expansion in patients with acute lupus. This raised the question of whether such precursors could be subject to immune tolerance. To address this, we generated a site-directed knock-in (KI) mouse line, G9gl, which carries germline-reverted sequences of the VH–DH–JH and Vκ–Jκ regions of patient-derived, high-affinity anti-dsDNA antibodies. G9gl heterozygous mice had a reduced number of peripheral B cells, only 27% of which expressed G9gl B cell receptor (BCR). The remaining B cells harbored non-KI allele-derived immunoglobulin heavy (IgH) chains or fusion products of upstream mouse VH and the KI gene, suggesting that receptor editing through VH replacement occurred in a large proportion of B cells in the KI mice. G9gl BCR-expressing B cells responded to ssDNA but not dsDNA, and exhibited several anergic phenotypes, including reduced surface BCR and shortened life span. Further, G9gl B cells were excluded from germinal centers (GCs) induced by several conditions. In particular, following immunization with methylated bovine serum albumin-conjugated bacterial DNA, G9gl B cells occurred at a high frequency in memory B cells but not GC B cells or plasmablasts. Thus, multiple tolerance checkpoints prevented low-affinity precursors of pathogenic anti-dsDNA B cells from undergoing clonal expansion and affinity maturation in GCs.

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Gene expression analysis of high-glucose stimulated human mesangial cells and tubular epithelial cells

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Expression data from primary mesangial cells stimulated with DNA and RNA ligands

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Gene expression analysis of high-glucose stimulated human mesangial cells and tubular epithelial cells

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Transcription profiling of mouse primary mesangial cells stimulated with DNA and RNA ligands

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