|
| Status |
Public on Jan 04, 2019 |
| Title |
CD8+ T eff, P. blood 3 |
| Sample type |
RNA |
| |
|
| Source name |
CD8+ Teff_P. blood
|
| Organism |
Mus musculus |
| Characteristics |
strain background: C57Bl/6 m-act OVA
gender: female
age: 8-16 weeks
tissue: peripheral blood
cell type: CD8+ T effector (Teff) cells
|
| Treatment protocol |
Lethal acute graft versus host disease (aGVHD) was induced in healthy m-act OVA C57Bl/6 female mice by allogeneic bone marrow transplantation (BMT). On the day of BMT (BMT 0) recipient m-act OVA mice (C57Bl/6-Tg(CAG-OVA)-916Jen/J) received 11Gy total body irradiation (TBI) in two split doses (5,5 Gy each), 3 hours apart, using a linear accelerator (1,07Gy/min). Irradiated animals were transplanted with 3x10^6 bone allogeneic marrow cells and 1.5x10^7 splenocytes via retroorbital injection, under ketamine-xylazine anesthesia from donor OT-I mice (C57BL/6-Tg(TcraTcrb) 1100Mjb/J). Graft OT-I CD8+ T cells were obtained from CD45.1 animals, while all other graft cells and recipients carried the CD45.2 allele. Four days after BMT (BMT+4) recipients were euthanized for sample retrieval.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
On the BMT+4 day transplanted recipients were sacrificed to obtain the small intestine, lungs, liver and peripheral blood. Grafted CD8+ T cells were labeled with anti-CD45.1 antibodies (Miltenyi Biotec), anti-Biotin Microbeads were added, and grafted CD45.1 CD8+ OT-I T cells were retrieved by positive selection using the AutoMACS Pro system (Miltenyi Biotec). Total RNA was extracted from MACS-separated T cells using the RNeasy Plus Micro Kit (Qiagen) following the manufacturer's recommendations. RNA yield was measured and integrity checked on an Agilent 2100 Bioanalyzer (Agilent Technologies).
|
| Label |
Cy3
|
| Label protocol |
Two rounds of linear RNA amplificaton were performed by the Arcturus RiboAmp HS PLUS, Cy3 Kit (Life Technologies) according to the manufacturer's instructions. Dye incorporation and cRNA yield were checked with the NanoDrop ND-1000 Spectrophotometer.
|
| |
|
| Hybridization protocol |
Amplified samples were hybridized to 4x44k mouse whole genome microarrays using the Gene Expression Hybridization Kit (Agilent)
|
| Scan protocol |
Samples were scanned on an Agilent Microarray Scanner using one color scan setting for 4x44k array slides
|
| Description |
13
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.7 (Agilent). Raw data were quantile normalized and further analyzed by Partek GS (Partek).
|
| |
|
| Submission date |
Jan 16, 2015 |
| Last update date |
Jan 04, 2019 |
| Contact name |
Nikolett Lupsa |
| E-mail(s) |
[email protected]
|
| Organization name |
Semmelweis University
|
| Department |
Genetics, Cell-and Immunbiology
|
| Lab |
MTA-SE Lendület Research Group
|
| Street address |
Nagyvárad tér 4
|
| City |
Budapest |
| State/province |
Pest |
| ZIP/Postal code |
1085 |
| Country |
Hungary |
| |
|
| Platform ID |
GPL11202 |
| Series (2) |
| GSE65043 |
Comparative analysis of mouse CD8+ Teff cells infiltrating distinct organ targets of aGVHD by gene expression profiling |
| GSE65045 |
Comparative analysis of murine CD8+ resident memory and effector T cells in various organ environments |
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