mRNA prime–boost evolves precursors toward VRC01-like broadly neutralizing antibodies in preclinical humanized mouse models
Data files
Mar 26, 2024 version files 149.23 KB
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200708_GT8_D14_post_prime.xlsx
26.83 KB
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200729_GT8_D36_post_prime.xlsx
23.95 KB
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200909_g28_D36_post_boost_.xlsx
18.27 KB
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200909_g5_D36_post_boost_.xlsx
16.63 KB
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210224_g28v2_D8_post_boost.xlsx
17.06 KB
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210304_g28v2_D16_post_boost.xlsx
25.79 KB
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210316_g28v2_D28_post_boost.xlsx
17.68 KB
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README.md
3.02 KB
Abstract
Germline-targeting (GT) protein immunogens to induce VRC01-class broadly neutralizing antibodies (bnAbs) to the CD4 binding site (CD4bs) of the HIV Envelope have shown promise in clinical trials. Here, we preclinically validated the mRNA-LNP delivery of one such immunogen, eOD-GT8, as a soluble self-assembling 60mer nanoparticle in humanized mouse models. In a model with three humanized B cell lineages bearing distinct VRC01-precursor B cell receptors (BCRs) with similar affinities for eOD-GT8, all lineages could be simultaneously primed and undergo diversification and affinity maturation with no sign of exclusionary competition. Boosts drove precursor B cell participation in germinal centers, the accumulation of somatic hypermutations, including in key VRC01-class positions, and affinity maturation to boost and native-like antigens in two of the three precursor lineages. We have preclinically validated a prime-boost regimen of soluble self-assembling nanoparticles delivered by mRNA-LNP, demonstrating that multiple lineages can be primed, boosted, and diversify along the bnAb pathway.
README: mRNA prime–boost evolves precursors toward VRC01-like broadly neutralizing antibodies in preclinical humanized mouse models
https://doi.org/10.5061/dryad.rn8pk0pk5
BCR Sequences from germinal center (GC) in CLK adoptively transferred mice after prime-boost.
Description of the data and file structure
BCR sequencing data for mutation analysis of heavy chains (HCs) and light chains (LCs) are from co-transferred mice bearing CLK19,CLK09 and CLK21 B cells, those mice were primed with eOD-GT8-60 mer mRNA-LNP and boosted with either g5- 60mer or g28-60 mer or g28v2-60mer mRNA-LNP.
GC B single cells from mice immunized at the different time points were sorted in 96-well plates by antigen-specific immunogen probes . mRNA of HCs and LCs were amplified from single cell for sanger sequencing. BCR sequences were processed using the IMGT/HighV-QUEST.
Data files include:
nucleotide sequences of heavy and light chain from mice at 14 days post-prime with eOD-GT8-60 mer mRNA-LNP
nucleotide sequences of heavy and light chain from mice at 36 days post-prime with eOD-GT8-60 mer mRNA-LNP
nucleotide sequences of heavy and light chain from mice at 36 days post-boost with g5-60 mer mRNA-LNP
nucleotide sequences of heavy and light chain from mice at 36 days post-boost with g28-60 mer mRNA-LNP
nucleotide sequences of heavy and light chain from mice at 8 days post-boost with g28v2-60 mer mRNA-LNP
nucleotide sequences of heavy and light chain from mice at 16 days post-boost with g28v2-60 mer mRNA-LNP
nucleotide sequences of heavy and light chain from mice at 28 days post-boost with g28v2-60 mer mRNA-LNP
Details of data sources and file information are shown as follows:
| run_date | prime | Boost | Time_point | file_tag |
|---|---|---|---|---|
| 20-07-08 | eOD-GT8-mRNA | none | D14-post-prime | 200708_GT8_D14_post_prime |
| 20-07-29 | eOD-GT8-mRNA | none | D36-post-prime | 200729_GT8_D36_post_prime |
| 20-09-09 | eOD-GT8-mRNA | g5-mRNA | D36-post-boost | 200909_g5_D36_post_boost |
| 20-09-09 | eOD-GT8-mRNA | g28-mRNA | D36-post-boost | 200909_g28_D36_post_boost |
| 21-02-24 | eOD-GT8-mRNA | g28v2-mRNA | D8-post-boost | 210224_g28v2_D8_post_boost |
| 21-03-04 | eOD-GT8-mRNA | g28v2-mRNA | D16-post-boost | 210304_g28v2_D16_post_boost |
| 21-03-16 | eOD-GT8-mRNA | g28v2-mRNA | D28-post-boost | 210316_g28v2_D28_post_boost |
Each file contains two excel sheets including both heavy chain(HC) and light chain(LC) sequences.HC and LC sequences are unpaired.
Code/Software
As described in the "BCR sequencing" section of Methods and Materials:
Briefly, sequences from immunized mice underwent quality control, mutations were aligned to the reference CLK19, CLK09 and CLK21 sequences and analyzed using Geneious software, IMGT/V-QUEST was used for Human Ig gene analysis.
Methods
BCR Sequences from mice after prime-boost were isolated by single cell sequencing, see Materials and Methods/ BCR sequencing section. The unpaired sequences of heavy and light chains are shown in excel sheets.