|
| Status |
Public on Sep 13, 2007 |
| Title |
PP164_infected_DC |
| Sample type |
RNA |
| |
|
| Source name |
Monocyte derived Dendritic Cells from healthy human subjects
|
| Organism |
Homo sapiens |
| Characteristics |
Dendritic Cells infected with H. ducreyi from pustule former, id 164
|
| Treatment protocol |
Dendritic Cells incubated with H.ducreyi
|
| Growth protocol |
Peripheral blood was obtained from the 3 PP and 3RR subjects several mo after they were infected a third time. CD14+ peripheral blood monocytes were isolated from PBMC by positive selection magnetic beads (Miltenyi Biotec, Auburn, CA) and grown in the presence of recombinant human (rh) IL-4 (1 ng/ml) and rhGM-CSF (0.2 ng/ml) (R&D Systems, Minneapolis, MN). After 7 d of culture, the cells were HLA-DR+, CD86+ CD40+, CD3-, CD14-, and CD19- by flow cytometry. DC were incubated with nonopsonized H. ducreyi for 90 min at an MOI of 30:1, washed to remove non-associated bacteria and incubated an additional 22.5 h.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNeasy® Mini Kit, according to the manufacturer's instructions.
|
| Label |
biotin
|
| Label protocol |
Biotinylated cRNA was prepared according to the Affymetrix 2-cycle protocol from 100 ng total RNA.
|
| |
|
| Hybridization protocol |
Following fragmentation, 15 microg of cRNA were mixed with spike controls and 2/3 was hybridized for 17 hr at 45C on GeneChip Human Genome 133 plus 2 Array. GeneChips were washed and stained in the Affymetrix Fluidics Station per manufacturer's instructions.
|
| Scan protocol |
GeneChips were scanned using the Affymetrix GeneChip Scanner 3000
|
| Description |
Gene expression from Dendritic cells from human subject with pustule former phenotype infected with H. ducreyi.
|
| Data processing |
The data were analyzed with Microarray Suite version 5.0 (MAS 5.0) using Affymetrix default analysis settings and global scaling as normalization method. The trimmed mean target intensity of each array was set to 1000.
|
| |
|
| Submission date |
Aug 16, 2006 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Stanley M. Spinola |
| E-mail(s) |
sspinola@iupui.edu
|
| Organization name |
Indiana University School of Medicine
|
| Department |
Infectious Diseases
|
| Street address |
545 Barnhill Dr.
|
| City |
Indianapolis |
| State/province |
IN |
| ZIP/Postal code |
46202 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE5547 |
Host Susceptibility to H. ducreyi Infection is Associated with Unique Transcript Profiles in Tissue and Dendritic Cells |
|
| Relations |
| Reanalyzed by |
GSE64985 |
| Reanalyzed by |
GSE86362 |
| Reanalyzed by |
GSE119087 |
