Unknown,Transcriptomics,Genomics,Proteomics

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Transcriptomic signature of the CD24hiCD38hi transitional B cells associated with an immunoregulatory phenotype in renal transplant recipient.

ABSTRACT: The role of B cells after transplant regarding allograft rejection or tolerance has become a topic of major interest. Recently, in renal transplant recipients, a B cell signature characterized by the overexpression of CD19+CD38hiCD24hi transitional B cells has been observed in operationally tolerant patients and in Belatacept treated patients with significant lower incidence of donor specific antibodies. The phenotypic and functional characterization of these transitional B cells is far to be exhaustive. We present the first transcriptomic and phenotypic analysis associated with this phenotype. Three populations were studied and compared: (i) transitional CD24hiCD38hi (ii) CD24+CD38- and (iii) CD24intCD38int populations. Peripheral blood mononuclear cells were isolated from 5 healthy donors and CD20+ B cells were isolated by magnetic beads. Three B cells populations were then sorted by flow cytometry: CD19+CD24hiCD38hi (test), CD19+CD24+CD38- (control 1) and CD19+CD24intCD38int (control 2)

ORGANISM(S): Homo sapiens

SUBMITTER: Philippe Grimbert

PROVIDER: E-GEOD-76272 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:During a T cell response, naÃ¯ve CD8 T cells differentiate into effector cells. Subsequently, a subset of effector cells termed memory precursor effector cells (MPECs) further differentiates into functionally mature memory CD8 T cells. The transcriptional network underlying this carefully scripted process is not well understood. Here, we report that the transcription factor FoxO1 plays an integral role in facilitating effector to memory transition and functional maturation of memory CD4 and CD8 T cells. We find that FoxO1 is not required for differentiation of effector cells, but in the absence of FoxO1, memory CD8 T cells displayed features of scenescence and progressive attrition in polyfunctionality, which in turn led to impared recall responses and poor protective immunity. These data suggest that FoxO1 is essential for active maintenance of functional CD8 T cell memory and protective immunity. Under competing conditions in bone marrow Single-cell suspensions from splenocytes of eight samples WT (control) and FoxO1-/- (experimental) LCMV-immune mice were prepared using standard procedures. CD8 T cells were then isoloated using Thy1.2 (CD90.2) (30-H12) microbeads (Miltenyi Biotec). Cells were then stained with anti-CD8, anti-CD44 and Db/NP396 MHC class I tetramer. Activated (CD8+CD44hi), naive (CD8+CD44lo), and virus-specific CD8 T cells were sorted using FACSAria II instrument (BD Biosciences). The purity of the cells was >95%. Total RNA was extracted from the sorted cells by Trizol Reagent. RNA samples were reverse transcribed and Cy3-labeled cDNAs were hyrbidized to Agilent whole Mouse Genome Oligo Microarrays. Fluorscence signals were detected using Agilent's Microarray Scanner system, data was analyzed using the Rosetta Resolver gene expression data analysis system and genes with a fold change < and p-values <0.01 were identified. Microarray data discussed in the paper is focused on virus-specific memory CD8 T cells from samples WT_Tet_2 vs KO_Tet_2.

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Transcriptomic signature of the CD24hiCD38hi transitional B cells associated with an immunoregulatory phenotype in renal transplant recipient.

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