Project description:This series of microarray experiments monitored the gene expression profiles for monoclonal cell lines (derived from HEK-293 parental cell culture) with high (H1, H15, H24, H36, H39) or low (L3, L28, L29) levels of store-operated Ca2+ entry (SOCE). For selection of clones, HEK-293 cells were loaded with indo-1 and sorted by FACS on the basis of their cyclopiazonic acid (CPA)-stimulated Ca2+ entry. Monoclonal cell lines were selected from the sorted cells and their levels of SOCE confirmed by monitoring thapsigargin-stimulated Ba2+ entry. Total RNA was extracted from cells immediately after removal from their growth environment. RNA was processed and hybridized to the Affymetrix HG-U133A chip. Two parallel hybridizations were done for each RNA preparation from each monoclonal cell line or from the parental HEK-293 cell culture.
Project description:This series of microarray experiments monitored the gene expression profiles for monoclonal cell lines (derived from HEK-293 parental cell culture) with high (H1, H15, H24, H36, H39) or low (L3, L28, L29) levels of store-operated Ca2+ entry (SOCE). For selection of clones, HEK-293 cells were loaded with indo-1 and sorted by FACS on the basis of their cyclopiazonic acid (CPA)-stimulated Ca2+ entry. Monoclonal cell lines were selected from the sorted cells and their levels of SOCE confirmed by monitoring thapsigargin-stimulated Ba2+ entry. Total RNA was extracted from cells immediately after removal from their growth environment. RNA was processed and hybridized to the Affymetrix HG-U133A chip. Two parallel hybridizations were done for each RNA preparation from each monoclonal cell line or from the parental HEK-293 cell culture. Keywords: parallel sample
Project description:Calcium is a critical signaling molecule in many cell types including immune cells. The calcium-release activated calcium channels (CRAC) responsible for store-operated calcium entry (SOCE) in immune cells are gated by STIM family members functioning as sensors of Ca2+ store content in the endoplasmic reticulum. We investigated the effect of SOCE blockers BTP2 and CM4620 on human peripheral blood mononuclear cells (PBMCs) polyclonally-stimulated with the mitogen phytohemagglutinin (PHA).
Project description:Calcium is a critical signaling molecule in many cell types including immune cells. The calcium-release activated calcium channels (CRAC) responsible for store-operated calcium entry (SOCE) in immune cells are gated by STIM family members functioning as sensors of Ca2+ store content in the endoplasmic reticulum. We investigated the effect of SOCE blocker BTP2 on human peripheral blood mononuclear cells (PBMC) stimulated with the mitogen phytohemagglutinin (PHA).
Project description:Loss of function mutations of ORAI1 suppress store-operated Ca2+ entry (SOCE) and cause an immunodeficiency disorder called Ca2+ release-activated Ca2+ (CRAC) channelopathy. An infant patient who is compound heterozygous for His134Pro and Leu194Pro mutations in the ORAI1 gene had strongly reduced SOCE in response to T cell receptor stimulation. He suffered from a severe form of combined immunodeficiency (CID) resulting in fatal chronic cytomegalovirus infection. Single cell transcriptomics revealed an overall strong activation of the patient's T cells but attenuated CD8+ effector memory T cell function.
Project description:The purpose is to have a overview on how chronic dysfunction of store-operated calcium entry change transcription profiles of adult fly fat tissue. Methods:To achieve chronic dysfunction of store-operated calcium entry, we carried out the Temperature-caused RNAi Pulse Induction of Stim (29 celcius degree incubation, Stim-TRiPI, or Stim-TRIP in data file) in fat storage tissue of 6 days old adult male flies. At day 10 and 11 after Stim-TRiPI (Stim-TRIP), we isolated the fat tissues and carried out RNAseq analysis based gene-level read counts.
Project description:<p>Regulatory T cells (Tregs) expand during Mycobacterium tuberculosis (Mtb) infection and suppress T cell mediated control. Whether Mtb actively contributes to this process is unclear. Here, using a genome-wide mutant library, we show that expression of Mtb Rv1272c, an ATP-binding cassette transporter, increased under hypoxic condition, promotes Mtb survival in vivo by increasing lecithin import, followed by the production and release of linoleic acid. Linoleic acid released by infected macrophages promoted surface trafficking of the immune checkpoint molecule CTLA-4 in Tregs via the Ca²⁺ transporter ATP2a3. This in turn inhibited macrophage reactive oxygen species production and promoted Mtb survival inside macrophages. Rv1272c-induecd linoleic acid further promoted Mtb immune evasion via increasing CTLA-4 surface trafficking on Tregs in vivo. [AU please mention in vivo work on virulence as well] Mechanistically, linoleic acid interacts with ATP2a3 in Tregs and promotes mitochondria-associated endoplasmic reticulum (ER) membrane formation. This facilitates ER to mitochondrial Ca2+ transfer, depletion of Ca2+ in the ER, and triggers store-operated calcium entry, thus elevating cytosolic Ca2+ levels to increase Ca2+-dependent CTLA-4 surface trafficking in Tregs. These findings reveal that Mtb can use a metabolite to manipulate host responses and promote its intracellular survival.</p>
