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| Status |
Public on Mar 11, 2019 |
| Title |
In vivo microarray expression data from WT, Irf4-/- and checkpoint blockade treated Irf4-/- TEa CD4+ T cells |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by array
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| Summary |
Previously we found that Irf4-deficient T cells were dysfunctional and unable to reject heart allografts and checkpoint blockade treatment could reinvigorate dysfunctional Irf4-deficient T cells and enable them reject heart allografts. But shortly after the reinvigoration those T cells went back to dysfunctional state. TCR-transgenic TEa CD4+ T cells (B6 background) recognize a Balb/c I-Eα allopeptide presented by B6 antigen presenting cells, and were used to assess the effects of immune checkpoint blockades on Irf4-deficient alloreactive T cells. WT, Irf4-/- and checkpoint blockade treated Irf4-/- TEa CD4+ T cells were isolated from Balb/c heart transplanted B6 mice, followed by microarray analysis.
To define un-restored gene expressions in Irf4-deficient alloreactive T cells upon immune checkpoint blockades, we compared genes expression level among three groups. We revealed that checkpoint blockade restored the expression levels of the majority of wild-type T cell-expressed genes in Irf4-/- T cells, indicating the reinvigoration of Irf4-/- T cells. The remaining un-restored genes following checkpoint blockade, though minimal in number, may be responsible for the reinvigorated Irf4-/- T cells to become re-dysfunction.
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| Overall design |
TCR(Vα2+Vβ6+)CD45.2+CD4+ TEa cells were isolated from splenocytes of WT TEa or Irf4−/− TEa mice by a FACSAria flow cytometer (BD Biosciences). B6.SJL CD45.1+ congenic mice were adoptively transferred with either 5 x 10e6 CD45.2+ WT TEa or 5 x 10e6 CD45.2+ Irf4‒/‒ TEa cells on day -1, and transplanted with Balb/c hearts on day 0. CD45.1+ mice adoptively transferred with either CD45.2+ Irf4‒/‒ TEa cells were i.p. injected with 400 μg Rat IgG, or 200 μg anti-PD-L1 plus 200 μg anti-CTLA-4 (9D9) mAbs on days 0, 3, 5. On day 6, adoptively transferred CD45.2+ TEa cells were sorted from splenocytes of transplant recipients by the FACSAria flow cytometer. Total RNA was extracted from sorted cells with RNeasy mini kit (Qiagen), followed by microarray analysis. Two independent experiments for each group were performed.
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| Contributor(s) |
Wu J, Chen W, Zhang H |
| Citation(s) |
30468559 |
| Submission date |
Mar 13, 2018 |
| Last update date |
Apr 09, 2019 |
| Contact name |
Hedong Zhang |
| Organization name |
Houston Methodist Hospital Research Institute
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| Department |
Immunobiology & Transplant Science Center
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| Lab |
Wenhao Chen's Lab
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| Street address |
6670 Bertner Ave
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| City |
Houston |
| State/province |
TX |
| ZIP/Postal code |
77030 |
| Country |
USA |
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| Platforms (1) |
| GPL21163 |
Agilent-074809 SurePrint G3 Mouse GE v2 8x60K Microarray [Probe Name version] |
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| Samples (6)
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| GSM3039393 |
Irf4-/- CD4+ TEa T cells_Rat IgG treated_rep |
| GSM3039394 |
Irf4-/- CD4+ TEa T cells_checkpoint blockade treated |
| GSM3039395 |
Irf4-/- CD4+ TEa T cells_checkpoint blockade treated_rep |
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| Relations |
| BioProject |
PRJNA438072 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE111757_RAW.tar |
29.9 Mb |
(http)(custom) |
TAR (of TXT) |
| Processed data included within Sample table |
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