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| Status |
Public on Jan 10, 2020 |
| Title |
Transcriptional profilling of pathogen-specific T helper cells |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Introduction: We reported a in vitro pathogen-specific priming system to using pathogen lysate stimulated splenic CD11c+ DCs to differentiate pathogen-specific murine T helper cells. This method allowed us to compare side-by-side transcriptional profiles of bulk T helper cells that are specific to distinct pathogens and to study their composition and functionality.
Method: We stimulated mouse splenic CD11c+ DCs with 10ug/ml Lm or Cr lysate for 5hours. The DCs were subsequently washed and co-cultured at 1:5 ratio to purified, CFSE-labeled CD62L+CD44- mouse naïve CD4 T cells. At day-7, CD90+CFSE- (Pathogen-specific Th cells) or CD90+CFSE+ (naive T cells) were sorted from the same culture and were subjected to RNA isolation, library prep and subsequent sequencing.
Conclusion: We found that Lm- and Cr-specific Th cells have distinct transcriptomes and contained distinct Th subtypes. Lm-specific Th cells contained higher expressions of Th1-associated cytokines, while Cr-specific Th cells contained higher levels of Th17 associate cytokines and transcription factors. In addition, we have identified heterogeneity within the pathogen-specific Th cells, as they express various cytokines from multiple Th lineages. Lastly, we found a vast variety of genes differentially expressed between Lm- and Cr-specific Th cells, indicating that different pathogen-stimulated DCs denote distinct function and developmental cues to T cells during differentiation.
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| Overall design |
mRNA profiles of Lm- or Cr-specific Th cells were generated by Illumina sequencing in duplicate. CFSE+CD90+ naïve T cell from Lm and Cr condition cultures served as control.
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| Contributor(s) |
Pasare C, Gao Y |
| Citation(s) |
31967646 |
| NIH grant(s) |
| Grant ID |
Grant title |
Affiliation |
Name |
| R01 AI123176 |
INNATE MECHANISMS OF REGULATION OF MEMORY TH17 CELL RESPONSES |
CHILDREN'S HOSPITAL MEDICAL CENTER |
Chandrashekhar Pasare |
| R01 AI113125 |
ROLE OF BCAP IN REGULATING INFLAMMATION AND ADAPTIVE IMMUNITY |
CHILDREN'S HOSPITAL MEDICAL CENTER |
Chandrashekhar Pasare |
| R01 GM120196 |
Endocytic Trafficking and Cell Signaling in Models of ARC Syndrome |
UT SOUTHWESTERN MEDICAL CENTER |
Helmut J Kramer |
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| |
| Submission date |
Nov 26, 2019 |
| Last update date |
Apr 10, 2020 |
| Contact name |
Chandrashekhar Pasare |
| E-mail(s) |
chandrashekhar.pasare@cchmc.org
|
| Phone |
513-636-6656
|
| Organization name |
Cininnati Children's Hospital Medical Center
|
| Street address |
3333 Burnet Ave
|
| City |
Cincinnati |
| State/province |
OH |
| ZIP/Postal code |
45208 |
| Country |
USA |
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| Platforms (1) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA591896 |
| SRA |
SRP233309 |
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