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Diversity of V(D)J repertoire of germinal center (GC) and non-germinal center (NGC) B cells following W614A-3S peptide-KLH vaccination using Squalene emulsion and Alum formulation

ABSTRACT: We compared the efficacy of W614A-3S peptide formulated in either squalene emulsion (SQE) or in aluminum hydroxide (Alum) in BCR repertoire of peptide-specific B cells after 4 immunizations (at week 11 after the first intramuscular injection). The V(D)J sequencing of W614A-3S-specific BCR showed significant differences in BCR sequences and validates the dichotomy between adjuvant formulations.

ORGANISM(S): Mus musculus

PROVIDER: GSE198883 | GEO | 2022/03/24

REPOSITORIES: GEO

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Differential gene expression in W614A-3S peptide-specific B cell populations following W614A-3S peptide-KLH vaccination using Squalene emulsion and Alum formulation

Project description:W614A-3S peptide is a modified 3S motif of the HIV-gp41 (mutation W614A). We previously detected the presence of natural neutralizing antibodies directed against W614A-3S peptide (NAbs) in long-term non-progressor HIV+ patients. Here, we compared the efficacy of W614A-3S peptide formulated in either squalene emulsion (SQE) or in aluminum hydroxide (Alum) in inducing broadly-NAbs (bNAbs). Rabbit and mouse models were used to screen the induction of bNAbs following 4 immunizations. SQE was more efficient than Alum formulation in inducing W614A-3S-specific bNAbs with up to 67-93% of HIV strains neutralized. We then analyzed the quality of peptide-specific murine B cells by single-cell gene expression by quantitative Reverse Transcription-PCR and single-cell V(D)J sequencing. We found more frequent germinal center B cells in SQE than in Alum, albeit with a different gene expression profile. The V(D)J sequencing of W614A-3S-specific BCR showed significant differences in BCR sequences and validates the dichotomy between adjuvant formulations. All sixteen BCR sequences which were cloned were specific of peptide. Adjuvant formulations of W614A-3S-peptide-conjugated immunogen impact the quantity and quality of B cell immune responses at both the gene expression level and BCR sequence. We used a single-cell quantitative Reverse Transcription-PCR (qRT-PCR) approach to compare between the two formulations the quality of W614A-3S-specific B cell populations isolated from draining lymph nodes, one week after 2nd and 3rd immunizations (W3 and W5, respectively).

2022-03-19 | GSE198952 | GEO

Differential gene expression in W614A-3S peptide-specific B cell populations following W614A-3S peptide-KLH vaccination using Squalene emulsion and Alum formulation

Project description:Differential gene expression in W614A-3S peptide-specific B cell populations following W614A-3S peptide-KLH vaccination using Squalene emulsion and Alum formulation

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Diversity of V(D)J repertoire of germinal center (GC) and non-germinal center (NGC) B cells following W614A-3S peptide-KLH vaccination using Squalene emulsion and Alum formulation

Project description:Diversity of V(D)J repertoire of germinal center (GC) and non-germinal center (NGC) B cells following W614A-3S peptide-KLH vaccination using Squalene emulsion and Alum formulation

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RNA-seq of DRG neurons isolated from alum vs. alum+KLH immunized mice

Project description:Purpose: we have performed RNA-seq analysis of sensory neurons isolated from alum vs. alum+KLH immunized mice

2018-03-19 | GSE108428 | GEO

10x Genomics VDJ seqeuncing of IOMAgl germinal center B cells

Project description:This experiment was performed to identify somatic hypermutations in germinal center B cells of a mouse expressing the inferred germline of IOMA BCR after and a 5 step sequential immunization regimen.

2022-03-07 | GSE197951 | GEO

Alum Pickering Emulsion as Effective Adjuvant to Improve Malaria Vaccine Efficacy.

Project description: Not available

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Visualizing germinal center B cell dynamics at single cell transcriptome resolution

Project description:During a germinal center (GC) response, B cells diversify their immunoglobulin (Ig) genes by somatic hyper-mutation (SHM) and undergo clonal expansion and positive selection thereby enabling the generation of higher affinity antibodies. We have analyzed the genomic states underlying GC B cell dynamics by single cell RNA-Seq. Profiling of antigen specific GC B cells during the peak of the response, revealed four distinctive genomic states characterized by antigen presentation, apoptotic, mitochondrial and mitotic gene expression modules. Intersection of genomic states and Ig heavy-chain (Igh) class-switch trajectory suggested that mitochondrial machinery is utilized to support class-switch recombination (CSR). Furthermore, by analyzing the transcriptomes of B cells with varying affinity BCR sequences that assembled from single-cell RNA-seq data through a novel algorithm, we show that high affinity GC B cells manifest enhanced mitotic and BCR signaling transduction, but compromised antigen processing and presentation gene expression modules. Thus, we are developing a comprehensive framework of the genomic states and molecular pathways underlying GC B cell dynamics.

2018-03-10 | GSE79096 | GEO

BCR-Seq of Germinal Center B cells from mice infected with LCMV

Project description:To study how acute or chronic viral infection (LCMV-WE or Docile) affect somatic hypermutation and/or affinity maturation of Germinal Center B cells, wild-type or Bmi1-deleted mice were infected with either the acute (WE) versus chronic (Docile) strains of LCMV, GC B cells (CD19+IgDloCD95+CD38loCD138-) were isolated 14 days post-infection and thei BCR was sequenced.

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Alum and squalene-oil-in-water emulsion enhance the titer and avidity of anti-A? antibodies induced by multimeric protein antigen (1-11)E2, preserving the Igg1-skewed isotype distribution.

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B cell intrinsic TBK1 is essential for germinal center formation during infection and vaccination

Project description:Germinal center (GC) is a site where somatic hypermutation and clonal selection are coupled for antibody affinity maturation against infections. However, how GCs are formed and regulated is incompletely understood. Here, we identified an unexpected role of Tank-binding kinase-1 (TBK1) as a crucial B-cell-intrinsic factor for GC formation. Using immunization and malaria infection models, we show that TBK1-deficient B cells failed to form GC despite normal Tfh differentiation, although some malaria-infected B-cell-specific TBK1-deficient mice could survive by GC-independent mechanisms. Mechanistically, TBK1 phosphorylation elevates in B cells during GC differentiation and regulates the balance of IRF4/BCL6 expression by limiting CD40 and BCR activation through non-canonical NF-κB and AKTT308 signaling. In the absence of TBK1, CD40 and BCR signaling synergistically enhanced IRF4 expression in Pre-GC leading to BCL6 suppression, therefore failed to form GC. As a result, memory B cells generated from TBK1-deficient B cells fail to confer sterile immunity upon re-infection, suggesting that TBK1 determines B cell fate to promote long-lasting humoral immunity.

2021-12-12 | GSE188659 | GEO

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