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| Status |
Public on Sep 27, 2022 |
| Title |
Single-cell analysis reveals dynamics of human B cell differentiation and identifies novel B and antibody-secreting cell intermediates |
| Organism |
Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Differentiation of B cells into antibody-secreting cells (ASCs) is a key process to generate protective humoral immunity. A detailed understanding of the cues controlling ASC differentiation is important to devise strategies to modulate antibody formation. Here, we dissected differentiation trajectories of human naive B cells into ASCs using single-cell RNA sequencing. By comparing transcriptomes of B cells at different stages of differentiation from an in vitro model with ex vivo B cells and ASCs, we uncovered a novel pre-ASC population present ex vivo in lymphoid tissues. For the first time, a germinal-center-like population is identified in vitro from human naive B cells and possibly progresses into a memory B cell population through an alternative route of differentiation, thus recapitulating in vivo human GC reactions. Our work allows further detailed characterization of human B cell differentiation into ASCs or memory B cells in both healthy and diseased conditions.
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| Overall design |
Human naive B cells (CD19+CD27-IgG-IgD+) are sorted from healthy donor PBMCs and cultured on a feeder layer of human CD40L-expressing mouse fibroblasts and restimulated after six days. Cytokines typically expressed by follicular T cells (Tfh) (IL-21, IL-4) are added to mimic Tfh help for B cell differentiation. The dynamics of human naive B cell differentiation were investigated in detail by single-cell RNA sequencing. Cultured cells were single-cell sorted on day 11 based on the expression of CD27 and CD38 to obtain a faithful representation of cells at varying stages of B cell to ASC differentiation. Next, cells were processed for single-cell RNA sequencing using the SMARTseq2 method.
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| Contributor(s) |
Verstegen NJ, Pollastro S, Unger PA, Marsman C, Elias G, Jorritsma T, Streutker M, Baβler K, Händler K, Rispens T, Schultze JL, ten Brinke A, Beyer M, van Ham SM |
| Citation(s) |
36861964 |
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| Submission date |
Sep 27, 2022 |
| Last update date |
May 04, 2023 |
| Contact name |
Niels Verstegen |
| E-mail(s) |
niels.verstegen@gmail.com
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| Organization name |
Sanquin Research
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| Street address |
Plesmanlaan 125
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| City |
Amsterdam |
| ZIP/Postal code |
1066 CX |
| Country |
Netherlands |
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| Platforms (1) |
| GPL18573 |
Illumina NextSeq 500 (Homo sapiens) |
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| Samples (384)
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| Relations |
| BioProject |
PRJNA884677 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE214265_R1_expression.matrix.txt.gz |
27.0 Mb |
(ftp)(http) |
TXT |
| GSE214265_R1_metadata.csv.gz |
36.4 Kb |
(ftp)(http) |
CSV |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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