Project description:We analyzed the transcriptome (RNA-seq) of mouse plasma cells (PC) expressing a human Ig light chain from a patient suffering light chain deposition disease (LCDD) as compared to control plasma cells (WT and DH-LMP2A, the latter producing no complete immunoglobulins, only free Ig light chains). Ig light chains causing LCDD present structural peculiarities leading to their aggregation and deposition into tissues and organs, mainly the kidney. We sought to determine if plasma cells producing these abnormal Ig could present specific phenotypes. The aim of the present study is to analyse the transcriptomic signature of plasma cells producing abnormal Ig free light chains.
Project description:We analyzed the transcriptome (RNA-seq) of glomeruli in a mouse model of light chain deposition disease (LCDD) as compared to control mice (WT and DH-LMP2A, the latter producing no complete immunoglobulins, only free Ig light chains). Kidney lesions in LCDD are due to the deposition of an abnormal monoclonal free Ig light chain and comprise progressive thickening of basement membranes (tubular and glomerular) and nodular glomerulosclerosis resembling the lesions observed in diabetic nephropathy. The aim of the present study is to analyse the transcriptomic changes at early steps of glomerulosclerosis.
Project description:Chip-chip from pro-B cells from Rag1KO mice for H3K27ac and RNA Pol II Identification of novel enhancers from Ig heavy and light chain loci Rag1KO pro-B epigenetic landscape at Ig heavy and light chain loci
Project description:Chip-chip from pro-B cells from Rag1KO mice for H3K27ac and RNA Pol II Identification of novel enhancers from Ig heavy and light chain loci
Project description:We used 454 sequencing to assess the repertoire of B cell subsets from bone marrow, spleen, and small intestinal lamina propria from two mouse strains. We used a RAG2-GFP reporter mouse strain (129Sve background) to isolate CD19+ RAG2+ B lineage cells from bone marrow and small intestinal lamina propria and total splenic B cells. We used 5' RACE to amplify cDNA libraries using primers specific for the mu constant region of IgH and the Ig kappa constant region. We also used this technique to analyze total B cell libraries from Swiss Webster germ-free mice to compare to littermate controls that were cohoused with regular specific pathogen free (SPF) mice for 7 days. Examination of the Ig kappa repertoire and IgH repertoire in RAG2+ bone marrow B lineage cells compared to RAG2+ small intestinal lamina propria B lineage cells or total splenic B cells. There are 8 (Ig kappa) or 4 (IgH) independent experiments comparing repertoires in RAG2-GFP mice. Each experiment in RAG2-GFP+ mice consisted of a pool of 8-12 mice. There are 3 experiments comparing germ-free to colonized mouse total B cell repertoires, each consisting of one mouse per condition.
Project description:We used 454 sequencing to assess the repertoire of B cell subsets from bone marrow, spleen, and small intestinal lamina propria from two mouse strains. We used a RAG2-GFP reporter mouse strain (129Sve background) to isolate CD19+ RAG2+ B lineage cells from bone marrow and small intestinal lamina propria and total splenic B cells. We used 5' RACE to amplify cDNA libraries using primers specific for the mu constant region of IgH and the Ig kappa constant region. We also used this technique to analyze total B cell libraries from Swiss Webster germ-free mice to compare to littermate controls that were cohoused with regular specific pathogen free (SPF) mice for 7 days.