title: B cell receptor induced Fc receptor-like 5 expression is mediated by multiple signaling pathways converging on NF-κB and NFAT.
abstract: Fc receptor-like (FCRL) proteins are novel regulators of the B cell response to antigen. Human FCRL5 binds intact IgG and modifies the strength of antigen receptor (BCR) signaling. Altering FCRL5 expression could therefore regulate the B cell response to antigen. In this study, we found that FCRL5 expression is induced specifically upon BCR stimulation and dissected the molecular mechanism. FCRL5 mRNA and cell surface protein expression required prolonged BCR stimulation and de novo protein synthesis. Using chemical inhibitors and activators, we identified roles for several signaling pathways, indicating a complex mechanism. Specifically, the PI3K/AKT, JNK, PKC and IKK2-dependent classical NF-κB pathways were involved in induced FCRL5 expression. Furthermore, induced FCRL5 expression required elevation of intracellular Ca(++) and was partially blocked by cyclosporine A, a calcineurin inhibitor. The importance of the transcription factors NF-κB, NFAT and CREB-binding protein was revealed based on sensitivity to inhibitors. Using reporter gene assays, we showed that the core FCRL5 promoter was sufficient to drive induced gene expression. Mutations of two predicted NF-κB sites or an NFAT site in the core promoter abrogated induced gene expression, suggesting direct regulation of the FCRL5 gene by NF-κB and NFAT. In support, we detected binding of NF-κB and NFAT family proteins to oligonucleotides corresponding to the predicted sites. We propose that the identified intricate mechanism serves to ensure that FCRL5 is expressed on B cells at a precise time following antigen encounter, with potential implications regarding regulation of the B cell response.
authors.affiliation: Office of Biotechnology Products, Center for Drug Evaluation and Research, US Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20993, USA.
authors.forename: Bazarragchaa
authors.initials: B
authors.lastname: Damdinsuren
authors.affiliation: Office of Biotechnology Products, Center for Drug Evaluation and Research, US Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20993, USA.
authors.forename: Jessica
authors.initials: J
authors.lastname: Dement-Brown
authors.affiliation: Office of Biotechnology Products, Center for Drug Evaluation and Research, US Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20993, USA.
authors.forename: Huifang
authors.initials: H
authors.lastname: Li
authors.affiliation: Office of Biotechnology Products, Center for Drug Evaluation and Research, US Food and Drug Administration, 10903 New Hampshire Ave., Silver Spring, MD 20993, USA. Electronic address: mate.tolnay@fda.hhs.gov.
authors.forename: Mate
authors.initials: M
authors.lastname: Tolnay
mesh_terms: D002460:Cell Line; D024202:Electrophoretic Mobility Shift Assay; D005434:Flow Cytometry; D005786:Gene Expression Regulation / Q000276:immunology*; D006801:Humans; D008213:Lymphocyte Activation / Q000276:immunology; D016328:NF-kappa B / Q000276:immunology* / Q000378:metabolism; D050778:NFATC Transcription Factors / Q000276:immunology* / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D011947:Receptors, Antigen, B-Cell / Q000096:biosynthesis*; D011961:Receptors, Fc / Q000276:immunology* / Q000378:metabolism; D015398:Signal Transduction / Q000276:immunology*
chemical_list: C432367:FCRL5 protein, human; D016328:NF-kappa B; D050778:NFATC Transcription Factors; D011947:Receptors, Antigen, B-Cell; D011961:Receptors, Fc
keywords: B cell antigen receptor; B lymphocytes; Fc receptor-like; Signal transduction; Transcriptional regulation