In vivo detection of antigen-specific CD8+ T cells by immuno-positron emission tomography

Abstract

The immune system’s ability to recognize peptides on major histocompatibility molecules contributes to the eradication of cancers and pathogens. Tracking these responses in vivo could help evaluate the efficacy of immune interventions and improve mechanistic understanding of immune responses. For this purpose, we employ synTacs, which are dimeric major histocompatibility molecule scaffolds of defined composition. SynTacs, when labeled with positron-emitting isotopes, can noninvasively image antigen-specific CD8+ T cells in vivo. Using radiolabeled synTacs loaded with the appropriate peptides, we imaged human papillomavirus-specific CD8+ T cells by positron emission tomography in mice bearing human papillomavirus-positive tumors, as well as influenza A virus–specific CD8+ T cells in the lungs of influenza A virus–infected mice. It is thus possible to visualize antigen-specific CD8+ T-cell populations in vivo, which may serve prognostic and diagnostic roles.

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Fig. 1: SynTac design, validation and labeling strategy.
Fig. 2: Analysis of synTac specificity in vitro.
Fig. 3: Detection of bulk CD8+ T cells.
Fig. 4: PET–CT imaging with the HPV E7 synTac.
Fig. 5: PET–CT imaging with the IAV NP synTac.
Fig. 6: SynTac PET–CT imaging following different radiolabeling strategies.

Data availability

The original PET–CT DICOM files that support the findings of this study are available from the corresponding author upon reasonable request. They are not available from public repositories due to their large size (~1 GB per mouse). Source data are provided with this paper.

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Acknowledgements

A.W.W. is supported by the Arnold O. Beckman Postdoctoral Fellowship. R.W.C. is supported in part by funding from the Cancer Research Institute Irvington Postdoctoral Fellowship. M.R. was supported by the National Institutes of Health (grant 1K22CA226040-01). H.L.P. is supported by the Lustgarten Foundation (award ID 388167). The laboratory of H.L.P. receives financial support in the form of a sponsored research agreement from VIR in the general area of immunity to flu virus. We acknowledge Cue Biopharma for partial support of this work. The synTac technology was developed with support provided by the National Institutes of Health (U01GM094665, U54GM094662, R01AI145024 and R01CA198095 to S.C.A.). We acknowledge the Wollowick Family Foundation Chair in Multiple Sclerosis and Immunology (to S.C.A.) and Janet & Martin Spatz and the Helen & Irving Spatz Foundation. Additional support provided by the Albert Einstein Macromolecular Therapeutics Development Facility, the Einstein-Rockefeller-CUNY Center for AIDS Research (P30AI124414) and the Albert Einstein Cancer Center (P30CA013330).

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Authors

Contributions

A.W.W., S.H.Z., E.L.Z., S.C.A. and H.L.P. conceived of and designed experiments. S.H.Z., R.J.C., R.D.S., S.J.G. and S.C.A. designed and assembled the plasmids encoding the heavy and light chains of the synTacs. A.W.W., E.L.Z. and M.R. performed preliminary HPV, IAV and PET studies, respectively. A.W.W., S.H.Z., E.L.Z., S.C.K., R.W.C., M.R., S.J.G., J.L.D., M.M., P.K.D. and A.B.P. performed experiments. A.W.W., S.H.Z., E.L.Z., S.C.A. and H.L.P. analyzed and interpreted data. A.W.W., S.H.Z., S.C.A. and H.L.P. drafted the manuscript. All authors reviewed and edited the manuscript.

Corresponding authors

Correspondence to Steven C. Almo or Hidde L. Ploegh.

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Competing interests

The synTac technology was developed in the laboratory of S.C.A. and was licensed to Cue Biopharma, Inc., in which he holds equity, receives royalties and serves as chair of its scientific advisory board. S.J.G. and R.D.S. receive royalties from Cue Biopharma, Inc. The laboratory of H.P. receives financial support in the form of a sponsored research agreement from VIR Biotechnology in the general area of immunity to flu virus, but H.L.P. has no equity stake in VIR.

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Supplementary Information

Supplementary Figs. 1–14, Tables 1–6 and Source Data for Supplementary Figs. 1 and 3.

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Source Data Fig. 1

Unmodified Coomassie-stained gel; unprocessed western blot.

Source Data Fig. 6

Unmodified Coomassie-stained gels.

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Woodham, A.W., Zeigler, S.H., Zeyang, E.L. et al. In vivo detection of antigen-specific CD8+ T cells by immuno-positron emission tomography. Nat Methods (2020). https://doi.org/10.1038/s41592-020-0934-5

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