Dataset Information

Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8⁺ T-cell response.

ABSTRACT: Antigen-adjuvant conjugation is known to enhance antigen-specific T-cell production in vaccine models, but scalable methods are required to generate site-specific conjugation for clinical translation of this technique. We report the use of the cell-free protein synthesis (CFPS) platform as a rapid method to produce large quantities (> 100 mg/L) of a model antigen, ovalbumin (OVA), with site-specific incorporation of p-azidomethyl-L-phenylalanine (pAMF) at two solvent-exposed sites away from immunodominant epitopes. Using copper-free click chemistry, we conjugated CpG oligodeoxynucleotide toll-like receptor 9 (TLR9) agonists to the pAMF sites on the mutant OVA protein. The OVA-CpG conjugates demonstrate enhanced antigen presentation in vitro and increased antigen-specific CD8⁺ T-cell production in vivo. Moreover, OVA-CpG conjugation reduced the dose of CpG needed to invoke antigen-specific T-cell production tenfold. These results highlight how site-specific conjugation and CFPS technology can be implemented to produce large quantities of covalently-linked antigen-adjuvant conjugates for use in clinical vaccines.

SUBMITTER: Weiss AM

PROVIDER: S-EPMC7973483 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8<sup>+</sup> T-cell response.

Weiss Adam M AM Ajit Jainu J Albin Tyler J TJ Kapoor Neeraj N Maroju Shilpa S Berges Aym A Pill Lucy L Fairman Jeff J Esser-Kahn Aaron P AP

Scientific reports 20210318 1

Antigen-adjuvant conjugation is known to enhance antigen-specific T-cell production in vaccine models, but scalable methods are required to generate site-specific conjugation for clinical translation of this technique. We report the use of the cell-free protein synthesis (CFPS) platform as a rapid method to produce large quantities (> 100 mg/L) of a model antigen, ovalbumin (OVA), with site-specific incorporation of p-azidomethyl-L-phenylalanine (pAMF) at two solvent-exposed sites away from immu ...[more]

PMID: 33737644

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Project description:BackgroundA goal of cancer vaccines is to induce strong T cell responses to tumor antigens, but the delivery method, schedule, and formulation of cancer vaccines have not yet been optimized. Adjuvants serve to increase the immune response against vaccine antigens. However, little is known about the impact of adjuvants plus antigen and their delivery schedule on the immunologic milieu in the vaccine-site microenvironment (VSME). We hypothesized that antigen processing and presentation may occur directly in the VSME, that adding the toll-like receptor 3 (TLR3) agonist polyICLC (pICLC) would enhance markers of immune activation, and that the immune signatures would be enhanced further by repeated vaccination in the same skin site rather than after multiple vaccines in different skin locations.MethodsUsing RNA sequencing, we evaluated VSME biopsies from patients undergoing subcutaneous/intradermal peptide vaccination against melanoma, with incomplete Freund's adjuvant (IFA) with or without pICLC. Differential gene expression analyses and gene set enrichment analyses were performed using R. False discovery rate corrected p values <0.05 were considered significant.ResultsWe found that addition of peptide antigens to IFA enhanced antigen presentation pathways and a tertiary lymphoid structure gene-signature locally at the VSME. Addition of pICLC to IFA + peptide induced an immunologically favorable VSME 1 week after injection but had little impact on the VSME after three injections, compared with IFA + peptide alone. Repeated same-site injection of IFA + peptide antigens induced a VSME with more dendritic cell activation, Th1 dominance, and TLR adaptor protein gene expression than that induced by injections at different, rotating skin locations.ConclusionsThese data suggest that the vaccine-site itself may be a critically important location contributing to vaccine immunity rather than just the draining lymph node, that IFA induces a favorable VSME with TLR agonist being most beneficial early in the vaccine course, and that same-site injections lead to persistent stimulation of immune pathways that may be beneficial in eliciting antigen specific T cell expansion.

Dataset Information

Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8⁺ T-cell response.

Publications

Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8<sup>+</sup> T-cell response.

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Dataset Information

Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8+ T-cell response.

Publications

Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8<sup>+</sup> T-cell response.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets

Site-specific antigen-adjuvant conjugation using cell-free protein synthesis enhances antigen presentation and CD8⁺ T-cell response.