Development of a CD19 PET tracer for detecting B cells in a mouse model of multiple sclerosis

Background B cells play a central role in multiple sclerosis (MS) through production of injurious antibodies, secretion of pro-inflammatory cytokines, and antigen presentation. The therapeutic success of monoclonal antibodies (mAbs) targeting B cells in some but not all individuals suffering from MS highlights the need for a method to stratify patients and monitor response to treatments in real-time. Herein, we describe the development of the first CD19 positron emission tomography (PET) tracer, and its evaluation in a rodent model of MS, experimental autoimmune encephalomyelitis (EAE). Methods Female C57BL/6 J mice were induced with EAE through immunization with myelin oligodendrocyte glycoprotein (MOG(1-125)). PET imaging of naive and EAE mice was performed 19 h after administration of [Cu-64]CD19-mAb. Thereafter, radioactivity in organs of interest was determined by gamma counting, followed by ex vivo autoradiography of central nervous system (CNS) tissues. Anti-CD45R (B220) immunostaining of brain tissue from EAE and naive mice was also conducted. Results Radiolabelling of DOTA-conjugated CD19-mAb with Cu-64 was achieved with a radiochemical purity of 99% and molar activity of 2 GBq/mu mol. Quantitation of CD19 PET images revealed significantly higher tracer binding in whole brain of EAE compared to naive mice (2.02 +/- 0.092 vs. 1.68 +/- 0.06 percentage of injected dose per gram, % ID/g,p= 0.0173). PET findings were confirmed by ex vivo gamma counting of perfused brain tissue (0.22 +/- 0.020 vs. 0.12 +/- 0.003 % ID/g,p= 0.0010). Moreover, ex vivo autoradiography of brain sections corresponded with PET imaging results and the spatial distribution of B cells observed in B220 immunohistochemistry-providing further evidence that [Cu-64]CD19-mAb enables visualization of B cell infiltration into the CNS of EAE mice. Conclusion CD19-PET imaging can be used to detect elevated levels of B cells in the CNS of EAE mice, and has the potential to impact the way we study, monitor, and treat clinical MS.