Peptide mimic for influenza vaccination using nonnatural combinatorial chemistry
John J. Miles, … , David A. Price, Andrew K. Sewell
John J. Miles, … , David A. Price, Andrew K. Sewell
Published March 12, 2018
Citation Information: J Clin Invest. 2018;128(4):1569-1580. https://doi.org/10.1172/JCI91512.
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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

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Figure 4

The synthetic agonist specifically expands memory T cells that recognize processed and presented influenza peptide.

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The synthetic agonist specifically expands memory T cells that recognize...
(A and B) PBMCs from 2 HLA-A2+ healthy adults were incubated with GILGFVFTL (1 μM), gppqwnnpp (10 μM), retroinverted ltfvfglig (10 μM) peptides, or DMSO and cultured in vitro for 14 days (3 wells per condition, combined for staining). The cells were stained with HLA-A2–GILGFVFTL, –ALWGPDPAAA (preproinsulin, PPI), –CLGGLLTMV (EBV), and –NLVPMVATV (CMV) tetramers. The flow plots show GILGFVFTL tetramer-binding CD8+ T cells (numbers indicate percent frequency within the total CD8+ T cell population). Data are summarized graphically for all other culture conditions and tetramer specificities. (C and D) A third set of PBMCs was primed with GILGFVFTL or gppqwnnpp and stained with irrelevant (ALWGPDPAAA, PPI) and GILGFVFTL tetramers. (C) Administration of DMSO alone, with no peptide, was also performed as a control. Each line was incubated overnight alone or with CIR-WT (A2-), CIR-A2, CIR-A2/GAD65, or CIR-A2/M1, or with PHA (in duplicate). (D) Supernatants were harvested and activation quantified by MIP-1β ELISA. Associated flow plots can be found in Supplemental Figures 2 and 3. Errors from 2 replicates depict SEM.