CD16-IL15-CD33 Trispecific Killer Engager (TriKE) induces NK cell expansion, persistence, and myeloid blast antigen specific killing.

Abstract

Abstract NK cell infusions induce complete remissions in 30–50% of patients with refractory AML when combined with lymphodepleting chemotherapy and IL-2. NK cell therapy is limited by lack of antigen specificity and by IL-2 mediated induction of Tregs. To overcome these limitations we developed a 161533 trispecific killer engager (TriKE) molecule containing an anti-CD16 scFv, to engage NK cells, an anti-CD33 scFv, to engage AML targets, and an IL-15 molecule. We have shown previously that a 1633 bispecific killer engager (BiKE) is capable of creating an immunologic synapse between NK cells and AML targets leading to target killing. These molecules generate higher affinity interactions than natural CD16 binding to Fc portions of therapeutic antibodies, resulting in increased NK cell functionality. Addition of IL-15 in the TriKE molecule is meant to enhance NK cell priming, proliferation, and maintenance without inducing Tregs. Cytotoxicity assays and cytokine secretion assessment of NK cells incubated with CD33+ HL-60 target cells indicated that the TriKE induced greater NK cell activation than the BiKE or no molecule control. The TriKE was able to induce potent NK cell proliferation and survival when compared to the BiKE. Interestingly, when compared to recombinant IL-15 the TriKE induced less proliferation of T cells while inducing similar proliferation of NK cells from healthy donors, indicating specificity. The TriKE was capable of rescuing NK cell function of post-transplant patients. In vivo functionality was tested in an NSG xenogeneic model engrafted with HL-60-luc targets and NK cells. TriKE treatment resulted in reduced tumor burden and NK cell maintenance at day 21, displaying the immunotherapeutic potential of the molecule.