Design of TLR2-ligand-synthetic long peptide conjugates for therapeutic vaccination of chronic HBV patients

Synthetic long peptide (SLP) vaccination is a promising new treatment strategy for patients with a chronic hepatitis B virus (HBV) infection. We have previously shown that a prototype HBV-core protein derived SLP was capable of boosting CD4+ and CD8+ T cell responses in the presence of a TLR2-ligand in chronic HBV patients ex vivo. For optimal efficacy of a therapeutic vaccine in vivo, adjuvants can be conjugated to the SLP to ensure delivery of both the antigen and the co-stimulatory signal to the same antigen-presenting cell (APC). Dendritic cells (DCs) express the receptor for the adjuvant and are optimally equipped to efficiently process and present the SLP-contained epitopes to T cells. Here, we investigated TLR2-ligand conjugation of the prototype HBV-core SLP. Results indicated that TLR2-ligand conjugation reduced cross-presentation efficiency of the SLP-contained epitope by both monocyte-derived and naturally occurring DC subsets. Importantly, cross-presentation was improved after optimization of the conjugate by either shortening the SLP or by placing a valine-citrulline linker between the TLR2-ligand and the long SLP, to facilitate endosomal dissociation of SLP and TLR2-ligand after uptake. HBV-core SLP conjugates also triggered functional patient T cell responses ex vivo. These results provide an import step forward in the design of a therapeutic SLP-based vaccine to cure chronic HBV. •We generated TLR2-ligand HBV-SLP conjugates to ensure delivery of SLP and TLR2-ligand to the same DC upon vaccination.•TLR2-ligand HBV-SLP conjugates were less efficiently cross-presented by monocyte-derived and primary DC subsets in vitro.•Cross-presentation of TLR2-ligand HBV-SLP conjugates was improved by reducing SLP length.•Cross-presentation of TLR2-ligand HBV-SLP conjugates was improved by inclusion of a protease sensitive linker sequence.•TLR2-ligand HBV-SLP conjugates boosted chronic HBV patient-derived HBV-directed T cell responses ex vivo.