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NIH/NIAID On the resistance of Staphylococcus aureus to neutrophil microbicides

ABSTRACT: Staphylococcus aureus strain MW2 was exposed to the following neutrophil microbicides, hydrogen peroxide (H2O2), hypochlorous acid (HOCl) and azurophilic granule proteins. At the indicated time points, bacterial cultures were centrifuged and bacteria were lysed with RLT buffer (Qiagen) using a FastPrep system. Purification of MW2 RNA and subsequent preparation of labeled cDNA target was performed as described in Methods. Labeling of samples, GeneChip hybridization and scanning were performed according to standard Affymetrix protocols. Experiments were performed in triplicate, using three bacterial cultures from separate days for each treatment. These analyses provide an enhanced view of the mechanisms used by CA-MRSA to circumvent destruction by the human innate immune system. Keywords: time course with three treatments

ORGANISM(S): Rickettsia rickettsii Staphylococcus epidermidis RP62A Coxiella burnetii RSA 493 Granulibacter bethesdensis Staphylococcus epidermidis ATCC 12228 Chlamydia muridarum Chlamydia trachomatis D/UW-3/CX Staphylococcus haemolyticus JCSC1435 Borreliella burgdorferi B31 Staphylococcus aureus Chlamydia pneumoniae AR39 Staphylococcus aureus subsp. aureus MW2 Coxiella burnetii Chlamydia caviae GPIC

PROVIDER: GSE6716 | GEO | 2008/01/31

SECONDARY ACCESSION(S): PRJNA99043

REPOSITORIES: GEO

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NIH/NIAID On the resistance of Staphylococcus aureus to neutrophil microbicides

Project description:Staphylococcus aureus strain MW2 was exposed to the following neutrophil microbicides, hydrogen peroxide (H2O2), hypochlorous acid (HOCl) and azurophilic granule proteins. At the indicated time points, bacterial cultures were centrifuged and bacteria were lysed with RLT buffer (Qiagen) using a FastPrep system. Purification of MW2 RNA and subsequent preparation of labeled cDNA target was performed as described in Methods. Labeling of samples, GeneChip hybridization and scanning were performed according to standard Affymetrix protocols. Experiments were performed in triplicate, using three bacterial cultures from separate days for each treatment. These analyses provide an enhanced view of the mechanisms used by CA-MRSA to circumvent destruction by the human innate immune system. 48 samples total: MW2 with the following treatments, untreated, GP (azurophilic granule proteins), H2O2 (hydrogen peroxide) or HOCl (hypochlorous acid) at 4 time points in triplicate

2010-05-26 | E-GEOD-6716 | biostudies-arrayexpress

NIH/NIAID Neutrophil Response to A. phagocytophilum

Project description:Polymorphonuclear leukocytes (PMNs) were obtained from healthy individuals in accordance with protocols approved by the Institutional Review Board for Human Subjects at the University of Minnesota and the National Institute of Allergy and Infectious Diseases. PMNs (107) were combined on ice with live S. aureus (108) or with live or heat-killed A. phagocytophilum (bacteria isolated from 5x106 infected HL60 cells for a ratio of 1 infected HL60 cell: 2 PMNs, ~ 5-20 A. phagocytophilum: PMN) in wells of a 12-well tissue culture plate (pre-coated with 20% autologous normal human serum). Unstimulated control assays received either buffer (for S. aureus comparisons) or clarified HL60 lysate (for A. phagocytophilum comparisons). Plates were centrifuged at 350 x g for 8 min at 4oC to synchronize phagocytosis and incubated at 37 deg. C in a CO2 incubator for the indicated times. At the indicated times, tissue culture medium was aspirated from the plate and PMNs were lysed directly with RLT buffer (Qiagen, Valencia, CA). Purification of PMN RNA and subsequent preparation of labeled cRNA target was performed as described in Methods. Labeling of samples, hybridization of cRNA with HU133A oligonucleotide arrays (Affymetrix, Santa Clara, CA), and scanning were performed according to standard Affymetrix protocols ( http://www.affymetrix.com/pdf/expression_manual.pdf ). Experiments were performed in triplicate, using PMNs from three healthy individuals for each treatment. Keywords = HUMAN NEUTROPHILS BACTERIAL APOPTOSIS GENE REGULATION Keywords: ordered

2005-05-15 | GSE2405 | GEO

Gene Expression Profiling of Cellular Dose-Response to Hypochlorous Acid (HOCl)

2009-04-01 | GSE15457 | GEO

NIH/NIAID Chronic Granulomatous Disease Neutrophils

Project description:Polymorphonuclear leukocytes (PMNs) were obtained from healthy individuals (Healthy1, Healthy2, Healthy3, and Healthy4) or patients with X-linked chronic granulomatous disease (XCGD1, XCGD2, XCGD3, XCGD4, XCGD5, and XCGD6). The studies were performed in accordance with a protocol approved by the Institutional Review Board for Human Subjects, National Institute of Allergy and Infectious Diseases, and the Institutional Review Board for Human Subjects at the University of Iowa. All studies were conducted according to Declaration of Helsinki principles. PMNs (107) were combined with or without IgG and C3bi-coated latex beads (8 x 107) in wells of a 12-well tissue culture plate (pre-coated with 20% normal human serum) and centrifuged at 350 x g for 8 min at 4oC to synchronize phagocytosis. For the purpose of these studies, activated or "Stimulated" PMNs are defined as those that have been stimulated by phagocytosis of IgG- and C3bi-coated latex beads. "Control" PMNs are defined as resting cells or those left unstimulated throughout the time-course. Following centrifugation, plates were incubated at 37 deg. C in a CO2 for the indicated times. At the indicated times, tissue culture medium was aspirated from the plate and PMNs were lysed directly with RLT buffer (Qiagen, Valencia, CA). Purification of PMN RNA and subsequent preparation of labeled cRNA target (12 g) was performed as described in Methods. Labeling of samples, hybridization of cRNA with Hu95Av2 oligonucleotide arrays (Affymetrix, Santa Clara, CA), and scanning were performed according to standard Affymetrix protocols. Gene expression in healthy control and XCGD individuals was always compared at the same time points after phagocytosis Keywords = HUMAN PMN CHRONIC GRANULOMATOUS DISEASE NEUTROPHILS Keywords: parallel sample

2003-12-31 | GSE935 | GEO

NIH/NIAID GM-CSF Modulates Global Changes in Neutrophil Gene Expression

Project description:PMNs were isolated from venous blood of healthy individuals in accordance with a protocol approved by the Institutional Review Board for Human Subjects, National Institute of Allergy and Infectious Diseases. Human PMNs (107) were cultured in RPMI/H -/+ 100 ng/ml GM-CSF at 37oC and 5% CO2 for up to 24 h as indicated. At the indicated time points, tissue culture medium was aspirated from each well and PMNs were lysed with RLT buffer (Qiagen). Purification of PMN RNA and subsequent preparation of labeled cRNA target was performed as described in Methods. Labeling of samples, GeneChip hybridization (Hu95Av2 oligonuceotide microarrays) and scanning were performed according to standard Affymetrix protocols (see http://www.affymetrix.com/support/technical/manual/expression_manual.affx). Experiments were performed in triplicate, using PMNs from three healthy individuals for each treatment. These analyses allowed comparison of global gene expression in human neutrophils during spontaneous apoptosis with that in cells cultured with human GM-CSF. Keywords: time course

2005-10-07 | GSE2803 | GEO

Transcription profiling of Escherichia coli strain MG1655 response to 10 mgl-1 hypochlorous acid

Project description:Examining the response of chemostat cultured, steady-state, Escherichia coli MG1655 to 10 mg/l hypochlorous acid (HOCl)

2005-12-01 | E-MEXP-244 | biostudies-arrayexpress

RNA-Seq analysis of melanoma cells, B16F10, in the presence of hypochlorous acid (HOCl)

Project description:We evaluated the changes in the transcriptome of a melanoma cell line, B16F10, and spleen isolated murine T cells when in the presence of hypochlorous acid (HOCl) using RNA-sequencing technologies.

2020-12-30 | GSE164014 | GEO

The NIH-NIAID Filariasis Research Reagent Resource Center.

Project description: Not available

| S-EPMC3226539 | biostudies-literature

Panton-Valentine Leukocidin Does Not Impact Virulence of Community-associated MRSA

Project description:Panton-valentine leukocidin (PVL) has been linked to worldwide emergence of community-associated methicillin resistant Staphylococcus aureus (CA-MRSA) -- its role in virulence in unclear. Here we show that PVL had no effect on global gene expression of prominent CA-MRSA strains nor did it affect bacterial clearance from lungs, spleen and kidneys in a highly discriminatory rabbit bacteremia model. These findings negate a large body of epidemiological research that implicated PVL in CA-MRSA virulence. Keywords: mutant vs wild type in 2 different growth phases grown in 2 different medias Wild type USA 300 (strain SF8300), wild type USA 400 (strain MW2) were compared against their respective PVL isogenic knock out strains. Strains were compared at both mid-exponential and stationary phase and grown in both TSB and CCY to determine if PVL plays a role in gene regulation under these conditions.

2010-06-19 | E-GEOD-8677 | biostudies-arrayexpress

Transcription profiling of mouse RAW 264.7 cells treated with the oxidant hypochlorous acid (HOCl) - dose response

Project description:Hypochlorous acid (HOCl) is a potent oxidant that is produced endogenously in mammalian tissue by phagocytes. Exogenous exposure to HOCl also can occur following inhalation of chlorine gas. HOCl has been implicated as a source of oxidative stress associated atherosclerosis and other diseases. The purpose of this study was to identify dose-dependent transitions in cellular response to hypochlorous acid (HOCl), with a focus on understanding how various cellular defense and stress dose-responses overlap. Experiment Overall Design: RAW 264.7 cells were grown to 80-90% confluency in DMEM supplemented with supplemented with 10% fetal bovine serum (FBS), 10 mM HEPES, 100 U penicillin/ml, and 100 Âµg streptomycin/ml. Cells were treated with 0, 0.14, 0.35, 0.7, 1.4, 2.1, 2.8 or 3.5 mM HOCl in medium for 6 hr. Total RNA was isolated from cells with TRIzol according to manufacturer's instructions and then subjected to cleanup using RNase-Free DNase Set and RNeasy Mini kit. The resultant DNA-free RNA was diluted in RNase-free H2O and quantified by Nanodrop at 260 nm. The quality of RNA samples was confirmed using RNA Nano Chips with Agilent 2100 Bioanalyzer. RNA samples were stored at â70 oC until use. Experiment Overall Design: From 5 micrograms of total RNA, cDNA was synthesized using a one-cycle cDNA synthesis kit (Affymetrix Corp., Santa Clara, CA). cDNA was transcribed to cRNA which was then biotin-labeled using GeneChip IVT labeling kit. Fifteen micrograms of labeled cRNA were then hybridized to an Affymetrix Mouse Genome 430 2.0 Array at 45Â°C for 16 hr. Biological cRNA replicates (n = 3) were each hybridized to an individual array. After being washed using the GeneChip Fluidics Station 450, arrays were scanned using a GeneChip 3000 scanner and intensity values were extracted from the CEL file using Array Assist software (Stratagene, La Jolla, CA). Experiment Overall Design:

2009-04-12 | E-GEOD-15457 | biostudies-arrayexpress

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