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Series GSE147182 Query DataSets for GSE147182
Status Public on Mar 19, 2020
Title Dynamic Regulation of Tfh Clonal Selection During the Germinal Center Reaction
Organism Mus musculus
Experiment type Expression profiling by high throughput sequencing
Other
Summary The Germinal center is a dynamic microenvironment wherein B cells expressing high affinity antibody variants produced by hypermutation are selected for clonal expansion by limiting numbers of T follicular helper cells. Although a great deal is known about the mechanisms that control B cell selection in the germinal center, far less is understood about the clonal behavior of the T follicular helper cells that regulate this process. Here we report on the dynamic behavior of clones of T follicular helper cells during the germinal center reaction. We find that like germinal center B cells, T follicular helper cells undergo antigen dependent selection during the germinal center reaction resulting in differential proliferative expansion and contraction. Increasing the amount of antigen presented in the germinal center leads to increased T follicular cell division. Competition between T follicular helper cell clones is mediated by T cell receptor affinity for peptide-MHC ligand. Higher affinity T cells expanding preferentially in the germinal center show increased expression of genes downstream of the T cell receptor, genes required for metabolic reprogramming, cell division and cytokine production. These dynamic changes lead to dramatic remodeling of the functional T follicular cell repertoire during the germinal center reaction.
 
Overall design OTII-Fucci cells were adoptively transferred into C57Bl/6 mice that were immunized with NP-OVA and then boosted on day 7 with high and low affinity aDEC-APLs or nothing and analyzed after 18 hours. To examine the transcriptional programs associated with increased TCR affinity driven Tfh division we performed bulk mRNA sequencing. To determine whether there is dynamic redistribution of Tfh clones during a polyclonal immune response we followed clonotypes in the same mouse longitudinally by performing hemi-splenectomy on days 7 and 21 after immunization with NP-OVA. To ensure that we assayed T cells entering the spleen during the initial immune response we used mice that carry a tamoxifen inducible Cre in the CD62L locus and a tdTomato indicator (SellCreERT2 ROSAtdTmice). Tamoxifen injection into SellCreERT2 ROSAtdT reporter mice labels naïve T cells, but not Tfh cells and immunization recruits labeled naïve cells into the Tfh compartment. Naïve cells were labeled in reporter mice before immunization and following immunization tdTomato expressing Tfh were purified from hemisplectomized mice on days 7 and 21 after NP-OVA injection. Single cell RNA sequencing was carried out using Cellular Indexing of Transcriptomes and Epitopes by Sequencing (CITE-Seq) method.
 
Contributor(s) Merkenschlager J, Finkin S, Ramos V, Nowosad CR, Gazumyan A, Cipollac M, Oliveira TY, Nussenzweig MC
Citation(s) 37907454
Submission date Mar 18, 2020
Last update date Nov 16, 2023
Contact name Thiago Y Oliveira
E-mail(s) toliveira@rockefeller.edu
Organization name The Rockefeller University
Department Immunology, Virology and Microbiology
Lab Laboratory of Molecular Immunology
Street address 1230 YORK AVE
City NEW YORK
State/province New York
ZIP/Postal code 10065
Country USA
 
Platforms (2)
GPL19057 Illumina NextSeq 500 (Mus musculus)
GPL24247 Illumina NovaSeq 6000 (Mus musculus)
Samples (30)
GSM4420017 B1: Bcell_aDEC_WT_LZ_GC_FUCCI
GSM4420018 B2: Bcell_aDEC_WT_LZ_GC_FUCCI
GSM4420019 B3: Bcell_aDEC_WT_LZ_GC_FUCCI
Relations
BioProject PRJNA613275
SRA SRP253270

Download family Format
SOFT formatted family file(s) SOFTHelp
MINiML formatted family file(s) MINiMLHelp
Series Matrix File(s) TXTHelp

Supplementary file Size Download File type/resource
GSE147182_Bcell_aDEC_LOW_LZ_GC_FUCCI_vs_Bcell_aDEC_NO_LZ_GC_FUCCI.xlsx 4.9 Mb (ftp)(http) XLSX
GSE147182_Bcell_aDEC_WT_LZ_GC_FUCCI_vs_Bcell_aDEC_LOW_LZ_GC_FUCCI.xlsx 4.9 Mb (ftp)(http) XLSX
GSE147182_Bcell_aDEC_WT_LZ_GC_FUCCI_vs_Bcell_aDEC_NO_LZ_GC_FUCCI.xlsx 4.9 Mb (ftp)(http) XLSX
GSE147182_Bcells_TPM_values.xlsx 5.2 Mb (ftp)(http) XLSX
GSE147182_RAW.tar 95.5 Mb (http)(custom) TAR (of CSV, MTX, TSV)
GSE147182_Tcell_aDEC_LOW_TOTAL_OTII_TFH_vs_Tcell_NO_TOTAL_OTII_TFH.xlsx 5.0 Mb (ftp)(http) XLSX
GSE147182_Tcell_aDEC_WT_TOTAL_OTII_TFH_vs_Tcell_NO_TOTAL_OTII_TFH.xlsx 5.0 Mb (ftp)(http) XLSX
GSE147182_Tcell_aDEC_WT_TOTAL_OTII_TFH_vs_Tcell_aDEC_LOW_TOTAL_OTII_TFH.xlsx 4.9 Mb (ftp)(http) XLSX
GSE147182_Tcells_TPM_values.xlsx 5.3 Mb (ftp)(http) XLSX
GSE147182_scRNASeq10x_report.html.gz 3.3 Mb (ftp)(http) HTML
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Raw data are available in SRA
Processed data provided as supplementary file

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