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| Status |
Public on May 19, 2023 |
| Title |
Full-length single-cell BCR sequencing paired with RNA sequencing reveals convergent responses to vaccination |
| Organisms |
Macaca mulatta; Homo sapiens |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Single-cell RNA sequencing can to resolve transcriptional features from large numbers of individual immune cells, but techniques capable of resolving the variable regions of B cell receptors (BCR) – defining features that confer antigen specificity to B cells – remain limited, especially from widely-used 3`-barcoded libraries. Here, we report a method that for recovering paired, full-length variable region sequences of the BCRs from 3`-barcoded single-cell whole transcriptome libraries. We first verified this method could produce accurate, full-length BCR sequences. We then applied this method to profile antigen-specific B cell responses elicited against the capsular polysaccharide of Streptococcus pneumoniae serotype 3 (ST3) by glycoconjugate vaccines in infant rhesus macaques. Using our method, we defined features of the BCR associated with specificity for the ST3 antigen and showed that these sequence characteristics are present in multiple vaccinated monkeys, indicating a convergent response to vaccination. These results demonstrate the utility of our method to resolve key features of the B cell repertoire and for profiling antigen-specific responses elicited by vaccination.
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| Overall design |
B cells of interest were isolated from PBMC using either fluorscence associated cell sorting or immunomagnetic selection and analyzed with the Seq-Well platform for single-cell RNA sequencing.
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| Contributor(s) |
Morgan DM, Zhang Y, Kim J, Murillo M, Singh S, Loschko J, Surendran N, Patil SU, Kanevsky I, Chorro L, Love JC |
| Citation missing |
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| Submission date |
May 19, 2023 |
| Last update date |
May 22, 2023 |
| Contact name |
Duncan Matthew Morgan |
| E-mail(s) |
dmmorgan@mit.edu
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| Organization name |
MIT
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| Department |
Koch Institute
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| Lab |
Love lab
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| Street address |
500 Main St
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| City |
CAMBRIDGE |
| State/province |
MA |
| ZIP/Postal code |
02139 |
| Country |
USA |
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| Platforms (2) |
| GPL24676 |
Illumina NovaSeq 6000 (Homo sapiens) |
| GPL27943 |
Illumina NovaSeq 6000 (Macaca mulatta) |
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| Samples (82)
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| Relations |
| BioProject |
PRJNA974349 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE232873_RAW.tar |
602.0 Mb |
(http)(custom) |
TAR (of CSV, TSV, TXT) |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data provided as supplementary file |
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