TY - JOUR
T1 - SpyTag/SpyCatcher display of influenza M2e peptide on norovirus-like particle provides stronger immunization than direct genetic fusion
AU - Lampinen, Vili
AU - Gröhn, Stina
AU - Soppela, Saana
AU - Blazevic, Vesna
AU - Hytönen, Vesa P.
AU - Hankaniemi, Minna M.
N1 - Funding Information:
This work received funding from Tampere University Graduate School (VL, SG, SS), Finnish Foundation for Technology Promotion (VL), Instrumentarium Science Foundation (SS) and Tampere Tuberculosis Foundation (VL, MH, SG). We also acknowledge Academy of Finland (#335870, MH), Cancer Foundation Finland (VH) and Sigrid Juselius Foundation (VH) for financial support. Acknowledgments
Publisher Copyright:
Copyright © 2023 Lampinen, Gröhn, Soppela, Blazevic, Hytönen and Hankaniemi.
PY - 2023
Y1 - 2023
N2 - Introduction: Virus-like particles (VLPs) are similar in size and shape to their respective viruses, but free of viral genetic material. This makes VLP-based vaccines incapable of causing infection, but still effective in mounting immune responses. Noro-VLPs consist of 180 copies of the VP1 capsid protein. The particle tolerates C-terminal fusion partners, and VP1 fused with a C-terminal SpyTag self-assembles into a VLP with SpyTag protruding from its surface, enabling conjugation of antigens via SpyCatcher. Methods: To compare SpyCatcher-mediated coupling and direct peptide fusion in experimental vaccination, we genetically fused the ectodomain of influenza matrix-2 protein (M2e) directly on the C-terminus of norovirus VP1 capsid protein. VLPs decorated with SpyCatcher-M2e and VLPs with direct M2 efusion were used to immunize mice. Results and discussion: We found that direct genetic fusion of M2e on noro-VLP raised few M2e antibodies in the mouse model, presumably because the short linker positions the peptide between the protruding domains of noro-VLP, limiting its accessibility. On the other hand, adding aluminum hydroxide adjuvant to the previously described SpyCatcher-M2e-decorated noro-VLP vaccine gave a strong response against M2e. Surprisingly, simple SpyCatcher-fused M2e without VLP display also functioned as a potent immunogen, which suggests that the commonly used protein linker SpyCatcher-SpyTag may serve a second role as an activator of the immune system in vaccine preparations. Based on the measured anti-M2e antibodies and cellular responses, both SpyCatcher-M2e as well as M2e presented on the noro-VLP via SpyTag/Catcher show potential for the development of universal influenza vaccines.
AB - Introduction: Virus-like particles (VLPs) are similar in size and shape to their respective viruses, but free of viral genetic material. This makes VLP-based vaccines incapable of causing infection, but still effective in mounting immune responses. Noro-VLPs consist of 180 copies of the VP1 capsid protein. The particle tolerates C-terminal fusion partners, and VP1 fused with a C-terminal SpyTag self-assembles into a VLP with SpyTag protruding from its surface, enabling conjugation of antigens via SpyCatcher. Methods: To compare SpyCatcher-mediated coupling and direct peptide fusion in experimental vaccination, we genetically fused the ectodomain of influenza matrix-2 protein (M2e) directly on the C-terminus of norovirus VP1 capsid protein. VLPs decorated with SpyCatcher-M2e and VLPs with direct M2 efusion were used to immunize mice. Results and discussion: We found that direct genetic fusion of M2e on noro-VLP raised few M2e antibodies in the mouse model, presumably because the short linker positions the peptide between the protruding domains of noro-VLP, limiting its accessibility. On the other hand, adding aluminum hydroxide adjuvant to the previously described SpyCatcher-M2e-decorated noro-VLP vaccine gave a strong response against M2e. Surprisingly, simple SpyCatcher-fused M2e without VLP display also functioned as a potent immunogen, which suggests that the commonly used protein linker SpyCatcher-SpyTag may serve a second role as an activator of the immune system in vaccine preparations. Based on the measured anti-M2e antibodies and cellular responses, both SpyCatcher-M2e as well as M2e presented on the noro-VLP via SpyTag/Catcher show potential for the development of universal influenza vaccines.
KW - cell mediated and humoral immunity
KW - conjugation
KW - fusion protein
KW - influenza
KW - norovirus
KW - norovirus (NoV)
KW - SpyCatcher
KW - virus-like particle
U2 - 10.3389/fcimb.2023.1216364
DO - 10.3389/fcimb.2023.1216364
M3 - Article
C2 - 37424789
AN - SCOPUS:85164191512
SN - 2235-2988
VL - 13
JO - Frontiers in Cellular And Infection Microbiology
JF - Frontiers in Cellular And Infection Microbiology
M1 - 1216364
ER -