Incorporation of a Novel CD16-Specific Single-Domain Antibody into Multispecific Natural Killer Cell Engagers With Potent ADCC.

Multispecific antibodies that bridge immune effector and tumor cells have shown promising preclinical and clinical efficacies. Here, we isolated and characterized novel llama single-domain antibodies (sdAbs) against CD16. One sdAb, NRC-sdAb048, bound recombinant human and cynomolgus monkey CD16 ectodomains with equivalent affinity (KD: 1 nM) but did not recognize murine CD16. Binding was similar for human CD16a expressed on NK cells and CD16b (NA2) expressed on neutrophils but dramatically weaker (KD: ∼6 µM) for the CD16b (NA1) allotype. The sdAb stained primary human peripheral blood NK cells. Irrespective of fusion orientation and linker length, bispecific sdAb-sdAb and sdAb-scFv dimers (anti-CD16/EGFR, anti-CD16/HER2, and anti-CD16/CD19) retained full binding affinity for each target, coengaged both antigens simultaneously, elicited ADCC against target antigen-expressing tumor cells in a reporter bioassay, and triggered target-specific activation and degranulation of primary NK cells as measured via interferon-γ and CD107a expression. These molecules may have applications in cancer immunotherapy.

Simultaneous engineering of natural killer cells for CAR transgenesis and CRISPR-Cas9 knockout using retroviral particles.

Natural killer (NK) cells are potent cytotoxic innate lymphocytes that can be used for cancer immunotherapy. Since the balance of signals from activating and inhibitory receptors determines the activity of NK cells, their anti-tumor activity can be potentiated by overexpressing activating receptors or knocking out inhibitory receptors via genome engineering, such as chimeric antigen receptor (CAR) transgenesis and CRISPR-Cas9-mediated gene editing, respectively. Here, we report the development of a one-step strategy for CRISPR-Cas9-mediated gene knockout and CAR transgenesis in NK cells using retroviral particles. We generated NK cells expressing anti-epidermal growth factor receptor (EGFR)-CAR with simultaneous TIGIT gene knockout using single transduction and evaluated the consequence of the genetic modifications in vitro and in vivo. Taken together, our results demonstrate that retroviral particle-mediated engineering provides a strategy readily applicable to simultaneous genetic modifications of NK cells for efficient immunotherapy.

Synthesis of Planar-Type ZnO Powder in Non-Nano Scale Dimension and Its Application in Ultraviolet Protection Cosmetics.

ZnO is one of the most widely used inorganic sunscreens, owing to its fine particle size and UV light shielding capability. However, powders at nanosizes can be toxic and cause adverse effects. The development of non-nanosized particles has been slow. The present work investigated synthesis methods of non-nanosized ZnO particles for ultraviolet protection application. By altering the starting material, KOH concentration, and input speed, the ZnO particles can be obtained in different forms, including needle type, planar type, and vertical wall type. Cosmetic samples were made by mixing different ratios of synthesized powders. The physical properties and the UV blockage efficacy of different samples were evaluated using scanning electron microscopy (SEM), X-ray diffraction (XRD), particle size analyzer (PSA), and ultraviolet/visible (UV/Vis) spectrometer. The samples with 1:1 ratio of needle-type ZnO and vertical wall-type ZnO exhibited superior light blocking effect owing to improved dispersibility and prevention of particle agglomeration. The 1:1 mixed sample also complied with the European nanomaterials regulation due to the absence of nanosized particles. With superior UV protection in the UVA and UVB regions, the 1:1 mixed powder showed potential to be used as a main ingredient in UV protection cosmetics.