Peptide mimic for influenza vaccination using nonnatural combinatorial chemistry
John J. Miles, Mai Ping Tan, Garry Dolton, Emily S.J. Edwards, Sarah A.E. Galloway, Bruno Laugel, Mathew Clement, Julia Makinde, Kristin Ladell, Katherine K. Matthews, Thomas S. Watkins, Katie Tungatt, Yide Wong, Han Siean Lee, Richard J. Clark, Johanne M. Pentier, Meriem Attaf, Anya Lissina, Ann Ager, Awen Gallimore, Pierre J. Rizkallah, Stephanie Gras, Jamie Rossjohn, Scott R. Burrows, David K. Cole, David A. Price, Andrew K. Sewell
John J. Miles, Mai Ping Tan, Garry Dolton, Emily S.J. Edwards, Sarah A.E. Galloway, Bruno Laugel, Mathew Clement, Julia Makinde, Kristin Ladell, Katherine K. Matthews, Thomas S. Watkins, Katie Tungatt, Yide Wong, Han Siean Lee, Richard J. Clark, Johanne M. Pentier, Meriem Attaf, Anya Lissina, Ann Ager, Awen Gallimore, Pierre J. Rizkallah, Stephanie Gras, Jamie Rossjohn, Scott R. Burrows, David K. Cole, David A. Price, Andrew K. Sewell
View: Text | PDF
Research Article Immunology Infectious disease

Peptide mimic for influenza vaccination using nonnatural combinatorial chemistry

Abstract

Polypeptide vaccines effectively activate human T cells but suffer from poor biological stability, which confines both transport logistics and in vivo therapeutic activity. Synthetic biology has the potential to address these limitations through the generation of highly stable antigenic “mimics” using subunits that do not exist in the natural world. We developed a platform based on D–amino acid combinatorial chemistry and used this platform to reverse engineer a fully artificial CD8+ T cell agonist that mirrored the immunogenicity profile of a native epitope blueprint from influenza virus. This nonnatural peptide was highly stable in human serum and gastric acid, reflecting an intrinsic resistance to physical and enzymatic degradation. In vitro, the synthetic agonist stimulated and expanded an archetypal repertoire of polyfunctional human influenza virus–specific CD8+ T cells. In vivo, specific responses were elicited in naive humanized mice by subcutaneous vaccination, conferring protection from subsequent lethal influenza challenge. Moreover, the synthetic agonist was immunogenic after oral administration. This proof-of-concept study highlights the power of synthetic biology to expand the horizons of vaccine design and therapeutic delivery.

Authors

John J. Miles, Mai Ping Tan, Garry Dolton, Emily S.J. Edwards, Sarah A.E. Galloway, Bruno Laugel, Mathew Clement, Julia Makinde, Kristin Ladell, Katherine K. Matthews, Thomas S. Watkins, Katie Tungatt, Yide Wong, Han Siean Lee, Richard J. Clark, Johanne M. Pentier, Meriem Attaf, Anya Lissina, Ann Ager, Awen Gallimore, Pierre J. Rizkallah, Stephanie Gras, Jamie Rossjohn, Scott R. Burrows, David K. Cole, David A. Price, Andrew K. Sewell

×

Figure 3

The synthetic agonist activates influenza virus matrix epitope-specific CD8+ T cells in the context of HLA-A2 and elicits polyfunctional outputs.

Options: View larger image (or click on image) Download as PowerPoint
The synthetic agonist activates influenza virus matrix epitope-specific ...
(A) Chromium release cytotoxicity assay using the GILGFVFTL-specific CD8 T cell clone GD with CIR and CIR-A2 target cells. Effector/target cell ratio of 10:1. Peptide was added directly to the wells at 10–5 M and incubated for 5 hours. Errors from 3 replicates depict SEM. (B) ALF3 cells were seeded at 250 cells per well for an IFN-γ ELISpot plate. Each condition used 10–4 M gppqwnnpp peptide with ALF3 alone (250 cells per well) or with CIR-WT (A2–), CIR-A2, CIR-A2/D227K/T228A (227/228), or CIR-A2/Q115E (QE). The CIRs were used at 100,000 cells per well. Errors from 3 replicates depict SEM. Unpaired, 1-tailed t test with P values displayed. (C) Clonal GD CD8+ T cells were incubated with C1R-A2 with the indicated concentrations of GILGFVFTL (left), gppqwnnpp (middle), or ELAGIGILTV (right). Five distinct effector functions (CD107a, IFN-γ, IL-2, MIP-1β, and TNF-α) were measured using flow cytometry. Bars depict the percentage of CD8+ T cells expressing each function. Pie charts showing function are displayed below each corresponding bar graph. The pie segments represent the fraction of CD8+ T cells expressing the number of functions indicated in the key.