Dataset Information

Gene expression profile of human monocytes treated with RPMI-1640 medium alone or with medium containing sMHC II (1 ug/ml) for three hours

ABSTRACT: We examined the effects of ligating CD4 expressed by primary human peripheral blood monocytes with soluble MHC Class II. 3 replicates are included. Fresh blood monocytes, obtained through negative selection via magnetic activated cell sorting (Miltenyi, Auburn, CA), from three separate individual donors were treated with RPMI-1640 medium alone or with medium containing sMHC II (1 ug/ml) for three hours. Cells were harvested and RNA purified using the RNeasy procedure (Qiagen). RNA was then analyzed by the UCLA Clinical Microarray Core Laboratory using the Human U133 Plus 2.0 Array (Affimetrix). Data was analyzed using Dchip software. Visual inspection did not find any potential outliers or suspect areas. Data across all groups were normalized using the PM/MM difference.

REANALYSIS of: E-GEOD-32939

ORGANISM(S): Homo sapiens

PROVIDER: E-GEOD-32939 | ExpressionAtlas | 2015-09-21

REPOSITORIES: ExpressionAtlas

ACCESS DATA

Publications

CD4 ligation on human blood monocytes triggers macrophage differentiation and enhances HIV infection.

Zhen Anjie A Krutzik Stephan R SR Levin Bernard R BR Kasparian Saro S Zack Jerome A JA Kitchen Scott G SG

Journal of Virology 20140618 17

A unique aspect of human monocytes, compared to monocytes from many other species, is that they express the CD4 molecule. However, the role of the CD4 molecule in human monocyte development and function is not known. We determined that the activation of CD4 via interaction with major histocompatibility complex class II (MHC-II) triggers cytokine expression and the differentiation of human monocytes into functional mature macrophages. Importantly, we determined that CD4 activation induces intrace ...[more]

PMID: 24942581

Similar Datasets

Project description:Transcript profiles of Postia placenta grown on media containing ball-milled aspen or ball-milled pine as the sole carbon source were analyzed. Array design was based on the DoE's Joint Genome Institute's gene models for P. placenta version 1. The research goal is to identtify genes essential for cellulose depolymerization. From a data set of 12,438 unique alleles, each NimbleGen (Madison, WI) array featured 10 unique 60mers per gene, all in triplicate. The dataset was manually annotated to include only the ‘best allelic model’ among CAZY-encoding genes. Total RNA was purified from medium containing ball-milled aspen or ball-milled pine as the sole carbon source. Three biological replicates per medium were used (6 separate arrays). RNA was converted to double-strand cDNA and labeled with the Cy3 fluorophore sample for hybridization to the Postia placenta MAD-698 whole genome expression array by Roche NimbleGen (Iceland). In brief, 10ug of total RNA was incubated with 1X first strand buffer, 10 mM DTT, 0.5mM dNTPs, 100 pM oligo T7 d(T)24 primer, and 200 units of SuperScript II (Invitrogen) for 60 min at 42°C. Second strand cDNA was synthesized by incubation with 1X second strand buffer, 0.2mM dNTPs, 0.07 units per ul DNA ligase (Invitrogen), 0.27 units per ul DNA polymerase I (Invitrogen), 0.013 units per ul RNase H (Invitrogen), at 16°C for 2 hours. Immediately following, 10 units T4 DNA polymerase (Invitrogen) was added for additional 5 minute incubation at 16°C. Double-stranded cDNA was treated with 27ng/ul of RNase A (EpiCenter Technologies) for 10 minutes at 37°C. Treated cDNA was purified using an equal volume of phenol:chloroform:isoamyl alcohol (Ambion), ethanol precipitated, washed with 80% ethanol, and resuspended in 20ul water. One ug of each cDNA sample was amplified and labeled with 1 unit per ul of Klenow Fragment (New England BioLabs) and 1 O.D unit of Cy3 fluorophore (TriLink Biotechnologies, Inc.) for 2 hours at 37°C. Array hybridization was carried out with 6ug of labeled cDNA suspended in NimbleGen hybridization solution for 17 hours at 42°C.