89Zr-labeled nivolumab for imaging of T-cell infiltration in a humanized murine model of lung cancer
Nivolumab is a human monoclonal antibody specific for programmed cell death-1 (PD-1), a negative regulator of T-cell activation and response. Acting as an immune checkpoint inhibitor, nivolumab binds to PD-1 expressed on the surface of many immune cells and prevents ligation by its natural ligands. Nivolumab is only effective in a subset of patients, and there is limited evidence supporting its use for diagnostic, monitoring, or stratification purposes.
89Zr-Df-nivolumab was synthesized to map the biodistribution of PD-1-expressing tumor infiltrating T-cells in vivo using a humanized murine model of lung cancer. The tracer was developed by radiolabeling the antibody with the positron emitter zirconium-89 (89Zr). Imaging results were validated by ex vivo biodistribution studies, and PD-1 expression was validated by immunohistochemistry. Data obtained from PET imaging were used to determine human dosimetry estimations.
The tracer showed elevated binding to stimulated PD-1 expressing T-cells in vitro and in vivo. PET imaging of 89Zr-Df-nivolumab allowed for clear delineation of subcutaneous tumors through targeting of localized activated T-cells expressing PD-1 in the tumors and salivary glands of humanized A549 tumor-bearing mice. In addition to tumor uptake, salivary and lacrimal gland infiltration of T-cells was noticeably visible and confirmed via histological analysis.
These data support our claim that PD-1-targeted agents allow for tumor imaging in vivo, which may assist in the design and development of new immunotherapies. In the future, noninvasive imaging of immunotherapy biomarkers may assist in disease diagnostics, disease monitoring, and patient stratification.
KeywordsNivolumab Programmed cell death 1 (PD-1) Positron emission tomography (PET) Immunotherapy Immune checkpoint inhibitor immunoPET
This work was supported, in part, by the University of Wisconsin - Madison, the National Institutes of Health (NIBIB/NCI 1R01CA169365, 1R01CA205101, 1R01EB021336, T32CA009206, T32GM008505, 5T32GM08349, P30CA014520), the National Science Foundation (DGE-1256259), the American Cancer Society (125246-RSG-13-099-01-CCE), the National Science Foundation of China (81401465, 51573096), and the Basic Research Program of Shenzhen (JCYJ20170412111100742, JCYJ20160422091238319).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed.
- 6.Yuzefpolskiy Y, Baumann FM, Penny LA, Kalia V, Sarkar S. Signaling through PD-1 on CD8 T cells is critical for antigen-independent maintenance of immune memory. J Immunol. 2016;196(1 Supplement):129.6-.6.Google Scholar
- 14.Gettinger SN, Horn L, Gandhi L, Spigel DR, Antonia SJ, Rizvi NA, et al. Overall survival and long-term safety of nivolumab (anti-programmed death 1 antibody, BMS-936558, ONO-4538) in patients with previously treated advanced non-small cell lung cancer. J Clin Oncol. 2015;33(18):2004–12. doi: 10.1200/JCO.2014.58.3708.CrossRefPubMedPubMedCentralGoogle Scholar
- 19.Schilbach K, Schick J, Wehrmann M, Wollny G, Simon P, Schlegel PG, et al. PD-1-PD-L1 pathway is involved in suppressing alloreactivity of heart infiltrating t cells during murine gvhd across minor histocompatibility antigen barriers. Transplantation. 2007;84(2):214–22. doi: 10.1097/01.tp.0000268074.77929.54.CrossRefPubMedGoogle Scholar
- 24.The 2007 Recommendations of the international commission on radiological protection. ICRP publication 103. Ann ICRP. 2007;37(2–4):1–332. doi: 10.1016/j.icrp.2007.10.003.
- 27.Cancer Facts & Figures 2016. American Cancer Society. 2016;Atlanta, GA.Google Scholar
- 31.Poluektova LY, Makarov E. Human peripheral blood lymphocyte reconstituion as a model of neuroinflammation associated with graft-versus-host disease. In: Xiong H, Gendelman HE, editors. Current laboratory methods in neuroscience research. New York: Springer; 2014. p. 487–90.Google Scholar