The use of allogeneic, pluripotent stem-cell-derived immune cells for cancer immunotherapy has been the subject of recent clinical trials. In Japan, investigator-initiated clinical trials will soon begin for ovarian cancer treatment using human leukocyte antigen (HLA)-homozygous-induced pluripotent stem cell (iPSC)-derived antiglypican-3 (GPC3) chimeric antigen receptor (CAR)-expressing natural killer/innate lymphoid cells (NK/ILC). Using pluripotent stem cells as the source for allogeneic immune cells facilitates stringent quality control of the final product, in terms of efficacy, safety and producibility. In this paper, we describe our methods for the stable, feeder-free production of CAR-expressing NK/ILC cells from CAR-transduced iPSC with clinically relevant scale and materials. The average number of cells that could be differentiated from 1.8-3.6 × 10 6 iPSC within 7 weeks was 1.8-4.0 × 10 9 . These cells showed stable CD45/CD7/CAR expression, effector functions of cytotoxicity and interferon gamma (IFN-γ) production against GPC3-expressing tumor cells. When the CAR-NK/ILC cells were injected into a GPC3-positive, ovarian-tumor-bearing, immunodeficient mouse model, we observed a significant therapeutic effect that prolonged the survival of the animals. When the cells were injected into immunodeficient mice during non-clinical safety tests, no acute systemic toxicity or tumorigenicity of the final product or residual iPSC was observed. In addition, our test results for the CAR-NK/ILC cells generated with clinical manufacturing standards are encouraging, and these methods should accelerate the development of allogeneic pluripotent stem cell-based immune cell cancer therapies.
K E Y W O R D Schimeric antigen receptor, GPC3, ILC, immunotherapy, iPSC, NK | 1479 UEDA Et Al.
The antifungal activity of six fractions derived from garlic was investigated in an in vitro system. Ajoene had the strongest activity in these fractions. The growth of both Aspergillus niger and Candida albicans was inhibited by ajoene at <20 ,ug/ml.
Semiconducting thin films consisting of regioregular poly(3-hexylthiophene) (RR-PHT) and poly(N-dodecylacrylamide) (pDDA) were constructed by the Langmuir-Blodgett (LB) technique. A mixture of RR-PHT and pDDA spread from a chloroform solution on a water surface forms a stable monolayer, which can be transferred onto solid substrates by the LB method, yielding a well-defined polymer LB film. Surface morphology studies of the LB film indicate that the RR-PHT is dispersed uniformly throughout the surface. The polymer thin film was chemically doped by contacting with FeCl3 acetonitrile solution, and a conductivity of 5.6 S/cm was achieved. Further, the LB film was utilized as the semiconducting film in the field-effect transistor (FET), and mobilities of 2.2 x 10(-4) and 4.4 x 10(-4) cm2 V(-1) s(-1) were obtained by analyzing the saturated and linear regions of the current-voltage characteristic, respectively.
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