RNA-seq of SFB-tetramer+ cells
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA718888 | ENA |
REPOSITORIES: ENA
Dataset's files
| Action | DRS | |||
|---|---|---|---|---|
| SRR14121887_1.fastq.gz | Fastqsanger.gz | |||
| SRR14121887_2.fastq.gz | Fastqsanger.gz | |||
| SRR14121888_1.fastq.gz | Fastqsanger.gz | |||
| SRR14121888_2.fastq.gz | Fastqsanger.gz | |||
| SRR14121889_1.fastq.gz | Fastqsanger.gz |
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Project description:We utilized H2-I-Ab tetramers to isolate CD4 T cells from mice to characterize the functional characteristics of ileal intraepithelial T cells and their lymphoid counterparts in the corresponding draining lymph nodes (mLN). Non-CD4 T cells (B220+, CD11b+, CD11c+, F4/80+, CD8a+) were excluded before FACS-sorting of CD45+CD3+CD4+ tetramer-specific T cells discriminated by double-positive staining with dual-labeled tetramer fluorophores. We recovered tetramer+ and tetramer- IELs and mLN T cells, and profiled them by single-cell RNA and TCR sequencing.
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Project description:We report that adhesion of microbes to intestinal epithelial cells is a critical cue for Th17 induction. SFB colonized in the intestine of mice can adhere to mouse small intestinal epithelial cells and induce intestinal Th17 cells. However, SFB colonized in rats cannot adhere to mouse intestinal epithelial cells and induce Th17 cells. Likewise, Citrobacter rodentium (WT) can adhere to mouse colonic epithelial cells and induce Th17 cells, but non-adherent mutant of C. rodentium (Δeae) cannot induce Th17 cells. To examine the influence of adherent bacteria on intestinal epithelial cells, we performed RNA seq. Germ free mice were orally inoculated with M-SFB or R-SFB and total RNA was isolated from small intestinal epithelial cells 1 week after inoculation. Alternatively, germ free mice were orally inoculated with C. rodentium WT or eae mutant and total RNA was isolated from colonic epithelial cells 5 days after inoculation. The gene expression of small intestinal epithelial cells isolated from small intestine of germ free mice (2 mice), mice monocolonized with M-SFB (2 mice) or R-SFB (3 mice), and colon of germ free mice (3 mice), mice monocolonized C. rodentium WT (3 mice) or eae mutant (3 mice).
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