Aim:
Blinatumomab is an anti-CD19 bispecific T cell engager designed to bring together CD3+ T cells with CD19+ malignant B cells to enable synapse formation and disease eradication of B-ALL. We hypothesised that the proportion of T cells and their ability to bind to their targets may may be a biomarker of response. In this study, we assessed the effect of blinatumomab co-administered with reduced intensity chemotherapy in newly diagnosed B-ALL patients.
Method:
Patients with untreated Ph-ve B-ALL aged 40-65 years were enrolled on the ALLG sponsored ALL08 study (ACTRN12617000084381) and treated with alternating cycles of blinatumomab and methotrexate/cytarabine. Peripheral blood mononuclear cells (PBMC) were collected from patients (n=32) at baseline and after C1D28, and healthy age matched donors (n=5). Immune phenotyping of PBMC (31-plex, Cytek Aurora), live video microscopy of purified T cells co-incubated with CD19+ Raji target cells +/- blinatumomab (10 ng/ml), and whole PBMC synapse assays +/- blinatumomab (BD Fortessa) were performed. Analysis was conducted using FlowJo, IMARIS, and PRISM software.
Results:
After 1 cycle of blinatumomab treatment, CD19+ B cells decreased and naïve CD8+TIM3+ T cells increased in the blood of B-ALL patients. Live video microscopy demonstrated reduced kinetics and number of synapses formed after C1D28, indicating impaired blinatumomab-mediated T cell function. Further analysis using the whole PBMC synapse assay indicated fewer T cell multimers bound to endogenous B cells when the tumour burden exceeded 20% of circulating PBMC.
Conclusion:
Blinatumomab-driven synapse formation was reduced in patients with high B-ALL burden and resulted in increased naive CD8+ T cell frequency in the blood following one cycle of therapy. Despite ongoing T cell-B cell synapse formation there was a failure of clearance of residual B cells. Optimal blinatumomab therapeutic efficacy may require modifications of dosing schedule based on disease burden and demonstration of effective T cell engagement.