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| Status |
Public on Nov 30, 2021 |
| Title |
Effector Memory CD4 T cells engage the innate immune system through CD40 and TNFR to trigger cytokine storm and autoimmune pathology I |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Introduction: DCs sense and respond to a variety of pathogen-dependent and independent features. We found that DCs can directly sense the presence of immunological memory in the form of effector memory T cells. This activation by effector memory T cells led to a robust inflammatory signature in interacting DCs.
Method: Bone marrow derived DCs (BMDCs) were differentiated in the presence of GMCSF for 7 days. WT naïve T cells were polarized to Th0 (platebound anti-CD3 and anti-CD28 and soluble IL-2) for 5 days, followed by 2 days of rest in low IL-2 containing media to mimic generation of effector memory T cells. CD11c+ BMDCs were sorted via AutoMacs and activated with LPS (100ng/mL) or effector memory T cells (ratio of 1:4) in the presence or absence of soluble anti-CD3 (200ng/mL) for 3 hours. Cells were blocked with FC block and stained with flourophores to distinguish DCs from T cells. DCs were FACS sorted and lysed in Trizol.
Conclusion: BMDCs activated with LPS or effector memory T cells had a distict transcriptional response. When activated with effector memory T cells, BMDCs upregulated genes associated with TNFRSF signaling. BMDCs activated with LPS or effector memory T cells also shared certain transcriptional features of upregulated cytokine and chemokine expression.
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| Overall design |
mRNA profiles of BMDCs stimulated with LPS or effector memory Th0 cells via CD3 for 3 hours by Illumina sequencing in triplicate.
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| Contributor(s) |
Pasare C, McDaniel M, Chawla AS, Jain A, Meibers H |
| Citation(s) |
35061504 |
| |
| Submission date |
Sep 22, 2021 |
| Last update date |
Mar 01, 2022 |
| Contact name |
Chandrashekhar Pasare |
| E-mail(s) |
Chandrashekhar.pasare@cchmc.org
|
| Organization name |
Cincinnati Children's Hospital Medical Center
|
| Department |
Immunology
|
| Lab |
Chandrashekhar Pasare
|
| Street address |
250 Albert Sabin Way
|
| City |
Cincinnati |
| State/province |
Ohio |
| ZIP/Postal code |
45229 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (12)
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| Relations |
| BioProject |
PRJNA765348 |
| SRA |
SRP338240 |
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