Cross-presentation and genome-wide screening reveal candidate T cells antigens for a herpes simplex virus type 1 vaccine
Lichen Jing, … , Georges M.G.M. Verjans, David M. Koelle
Lichen Jing, … , Georges M.G.M. Verjans, David M. Koelle
Published January 3, 2012
Citation Information: J Clin Invest. 2012;122(2):654-673. https://doi.org/10.1172/JCI60556.
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Technical Advance Infectious disease

Cross-presentation and genome-wide screening reveal candidate T cells antigens for a herpes simplex virus type 1 vaccine

Abstract

Herpes simplex virus type 1 (HSV-1) not only causes painful recurrent oral-labial infections, it can also cause permanent brain damage and blindness. There is currently no HSV-1 vaccine. An effective vaccine must stimulate coordinated T cell responses, but the large size of the genome and the low frequency of HSV-1–specific T cells have hampered the search for the most effective T cell antigens for inclusion in a candidate vaccine. We have now developed what we believe to be novel methods to efficiently generate a genome-wide map of the responsiveness of HSV-1–specific T cells, and demonstrate the applicability of these methods to a second complex microbe, vaccinia virus. We used cross-presentation and CD137 activation–based FACS to enrich for polyclonal CD8+ T effector T cells. The HSV-1 proteome was prepared in a flexible format for analyzing both CD8+ and CD4+ T cells from study participants. Scans with participant-specific panels of artificial APCs identified an oligospecific response in each individual. Parallel CD137-based CD4+ T cell research showed discrete oligospecific recognition of HSV-1 antigens. Unexpectedly, the two HSV-1 proteins not previously considered as vaccine candidates elicited both CD8+ and CD4+ T cell responses in most HSV-1–infected individuals. In this era of microbial genomics, our methods — also demonstrated in principle for vaccinia virus for both CD8+ and CD4+ T cells — should be broadly applicable to the selection of T cell antigens for inclusion in candidate vaccines for many pathogens.

Authors

Lichen Jing, Jürgen Haas, Tiana M. Chong, Joseph J. Bruckner, Greg C. Dann, Lichun Dong, Joshua O. Marshak, Christopher L. McClurkan, Tori N. Yamamoto, Susanne M. Bailer, Kerry J. Laing, Anna Wald, Georges M.G.M. Verjans, David M. Koelle

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Figure 11

Schematic overview of the high-throughput T cell antigen discovery pathway.

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Schematic overview of the high-throughput T cell antigen discovery pathw...
See text for details. In this report, the microbe (left) was a virus, HSV-1, but the workflow can also be adapted to bacteria or parasites. A microbial ORF library is initially cloned in a flexible format and subcloned (not shown) into both a custom protein expression vector for CD4 assays and a custom transient expression vector for CD8 assays. The CD4 workflow (top) stimulates PBMCs with whole killed microbe and detects and isolates microbe-specific CD4+ T cells based on differential CD137 expression followed by expansion. The polyclonal CD4+ responder cells are then assayed against the protein library made in vitro from the secondary protein expression clone set. The CD8 workflow (bottom) uses cross-presentation by microbial antigen-laden DCs to stimulate CD8+ T cells from PBMCs. After CD137-based selection and expansion, effector cells are assayed for reactivity with panels of person-specific aAPCs made by co-transfection of Cos7 cells with participant HLA class I cDNA and the transient transfection microbial ORF set. The integrated assays assign each ORF a yes/no result for each participant for CD4+ and CD8+ T cells (turquoise).