ABSTRACT
Immunotherapy has emerged as a promising modality for cancer treatment. Dendritic cell immunoreceptor (DCIR), a C-type lectin receptor, is expressed mainly by dendritic cells (DCs) and mediates inhibitory intracellular signaling. Inhibition of DCIR activation may enhance antitumor activity. DCIR is encoded by CLEC4A in humans and by Clec4a2 in mice. Gene gun-mediated delivery of short hairpin RNA (shRNA) targeting Clec4a2 into mice bearing bladder tumors reduces DCIR expression in DCs, inhibiting tumor growth and inducing CD8+ T cell immune responses. Various oncolytic adenoviruses have been developed in clinical trials. Previously, we have developed Ad.LCY, an oncolytic adenovirus regulated by Oct4 and hypoxia, and demonstrated its antitumor efficacy. Here, we generated a Clec4a2 shRNA-expressing oncolytic adenovirus derived from Ad.LCY, designated Ad.shDCIR, aimed at inducing more robust antitumor immune responses. Our results show that treatment with Ad.shDCIR reduced Clec4a expression in DCs in cell culture. Furthermore, Ad.shDCIR exerted cytolytic effects solely on MBT-2 bladder cancer cells but not on normal NIH 3T3 mouse fibroblasts, confirming the tumor selectivity of Ad.shDCIR. Compared to Ad.LCY, Ad.shDCIR induced higher cytotoxic T lymphocyte (CTL) activity in MBT-2 tumor-bearing immunocompetent mice. In addition, Ad.shDCIR and Ad.LCY exhibited similar antitumor effects on inhibiting tumor growth. Notably, Ad.shDCIR was superior to Ad.LCY in prolonging the survival of tumor-bearing mice. In conclusion, Ad.shDCIR may be further explored as a combination therapy of virotherapy and immunotherapy for bladder cancer and likely other types of cancer.
ABSTRACT
According to the latest literature, it is difficult to measure the multiple important physical parameters inside a proton battery stack accurately and simultaneously. The present bottleneck is external or single measurements, and the multiple important physical parameters (oxygen, clamping pressure, hydrogen, voltage, current, temperature, flow, and humidity) are interrelated, and have a significant impact on the performance, life, and safety of the proton battery stack. Therefore, this study used micro-electro-mechanical systems (MEMS) technology to develop a micro oxygen sensor and a micro clamping pressure sensor, which were integrated into the 6-in-1 microsensor developed by this research team. In order to improve the output and operability of microsensors, an incremental mask was redesigned to integrate the back end of the microsensor in combination with a flexible printed circuit. Consequently, a flexible 8-in-1 (oxygen, clamping pressure, hydrogen, voltage, current, temperature, flow, and humidity) microsensor was developed and embedded in a proton battery stack for real-time microscopic measurement. Multiple micro-electro-mechanical systems technologies were used many times in the process of developing the flexible 8-in-1 microsensor in this study, including physical vapor deposition (PVD), lithography, lift-off, and wet etching. The substrate was a 50 µm-thick polyimide (PI) film, characterized by good tensile strength, high temperature resistance, and chemical resistance. The microsensor electrode used Au as the main electrode and Ti as the adhesion layer.
