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| Status |
Public on Aug 19, 2023 |
| Title |
Germinal centers continually recruit naïve T cells that alter the balance between follicular T helper and follicular regulatory T cells. |
| Organism |
Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Follicular helper T cells (T FH ) mediate B cell selection and clonal expansion in germinal centers (GCs), and follicular regulatory T cells (T FR ) prevent the emergence of self-reactive B cells and help to extinguish the reaction. Here we show that GC reactions continually recruit T cells from both the naïve conventional and naive thymic regulatory T cell (Treg) repertoires. In the early GC, newly recruited T cells develop into T FH , whereas cells entering during the contraction phase develop into T FR cells that contribute to GC dissolution. The T FR fate decision is associated with decreased antigen availability and is modulated by slow antigen delivery or mRNA vaccination. Thus, invasion of ongoing GCs by newly developing T FH and T FR from naïve conventional and Treg repertoires, helps remodel the GC based on antigen availability.
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| Overall design |
For single cell RNA sequencing, single cell suspensions were prepared from half-spleens of NP-OVA immunized SellCreERT2 ROSAtdT mice on day 7 and 21 after immunization. Samples were indexed with TotalSeqC (BioLegend) cell surface antibodies and CD4 + , CD62 low , CD44 hi , PD1 hi , CXCR5 high , tdTomato + Tfh cells were purified by flow cytometry, pooled and loaded onto a Chromium Controller (10x Genomics). Single-cell RNA-seq libraries were prepared using the Chromium Single Cell 5′ v2 Reagent Kit (10x Genomics) according to manufacturer’s protocol. Libraries were loaded onto an Illumina NextSeq with the mid-Output Kit (150 paired end) for V-D-J analysis or NOVAseq for single cell gene expression. Hashtag indexing was used to demultiplex the sequencing data and generate gene- barcode matrices, respectively.
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| Contributor(s) |
Merkenschlager J, Bernz RM, Ramos V, Uhlig M, MacLean AJ, Nowosad CR, Oliveira TY, Nussenzweig MC |
| Citation(s) |
37907454 |
| |
| Submission date |
Aug 14, 2023 |
| Last update date |
Nov 16, 2023 |
| Contact name |
Thiago Y Oliveira |
| E-mail(s) |
toliveira@rockefeller.edu
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| Organization name |
The Rockefeller University
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| Department |
Immunology, Virology and Microbiology
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| Lab |
Laboratory of Molecular Immunology
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| Street address |
1230 YORK AVE
|
| City |
NEW YORK |
| State/province |
New York |
| ZIP/Postal code |
10065 |
| Country |
USA |
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| Platforms (2) |
| GPL19057 |
Illumina NextSeq 500 (Mus musculus) |
| GPL24247 |
Illumina NovaSeq 6000 (Mus musculus) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA1005169 |
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