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| Status |
Public on Dec 29, 2023 |
| Title |
Loss of B cell Tolerance is TCR Dependent [amplicon-seq] |
| Organism |
Mus musculus |
| Experiment type |
Other
|
| Summary |
We tested orphan TCR autoreactivity using the peptide MHC-TCR chimeric receptor (MCR) co-culture system. In this system, cognate antigen recognition leads to TCR specific NFAT activation in MCR reporter cells expressing a mouse I-Ab MHC class II extracellular domain covalently linked to candidate peptides and an intracellular TCR signaling domain. We used mixed autoimmune bone marrow chimera spleens and kidneys as sources of cDNA to generate a transcriptome-wide library of natural autoantigen peptides . We cloned this cDNA-derived peptide (CDP) autoantigen library into the MCR retroviral backbone and transduced NFAT reporter cells to make a murine autoantigen MCR reporter library (MCR-Lib). We then used this library to screen orphan TCRs identified by scTCR-seq for autoreactivity.
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| Overall design |
Amplicon NGS was performed on PCR-amplified peptide sequences from MCR-Libraries following screening with orphan TCRs
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| Contributor(s) |
Akama-Garren EH, Carroll MC |
| Citation(s) |
38363829 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 AR074105 |
Evolution of autoreactive GC and epitope spreading in lupus |
CHILDREN'S HOSPITAL |
Michael Craig Carroll |
| F30 AI160909 |
Follicular helper T cells as drivers of epitope spreading |
HARVARD UNIVERSITY (MEDICAL SCHOOL) |
Elliot Hideki Akama-Garren |
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| |
| Submission date |
Apr 17, 2023 |
| Last update date |
Mar 29, 2024 |
| Contact name |
Elliot Akama-Garren |
| Organization name |
Harvard Medical School
|
| Street address |
200 Longwood Ave
|
| City |
Boston |
| State/province |
MA |
| ZIP/Postal code |
02115 |
| Country |
USA |
| |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (34)
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| Relations |
| BioProject |
PRJNA956588 |
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