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CD8+ T cells play a critical role during adaptive immune response, which often change locations and expand or contract in numbers under different states. In the past, many attempts to develop CD8+T cells that express luciferase in vivo have involved the use of viral transduction, which has drawbacks of hardly tracked via detection of luciferase signal in untouched natural states. Here, we generate a transgenic mouse model via CRISPR-mediated genome editing, C57BL/6-CD8a em(IRES-AkaLuci−2A-EGFP) knock-in mice(CD8a-Aka mice), as a novel tool for non-invasive imaging of CD8+ T cells, which expressed a highly sensitive luciferase-Akaluciferase. Our study offers a convenient and robust tool for understanding fundamental CD8+ T cell biology in experimental applications and preclinical translational studies. • Generate C57BL/6-CD8a em(IRES-AkaLuc −2A-EGFP) knock-in mice. • Aka-luciferase expression offers superior sensitivity to track CD8+ T cells in vivo. • Higher Akaluciferase expression signals found in "hot" solid tumor model established in CD8a-Aka mice compared with "cold" ones. • Aka-luciferase expression in CD8a-Aka mice is responsive to checkpoint blockade in the murine MC38 colorectal carcinoma model. [ABSTRACT FROM AUTHOR]

Oncolytic virus can selectively recognize cancer cells, target tumors, and stimulate an oncolytic and immune response. Recombinant armed oncolytic vaccinia virus has emerged as an attractive tool in oncolytic virotherapy because it has tumor-specific cytotoxicity and serves as a vector to express immune genes. A novel thymidine kinase (TK) gene–deleted oncolytic vaccinia virus (named Δ TK -Armed-VACV) armed with anti-human-programed cell death-1 protein (PD-1) antibody and anti-human-tumor necrosis factor receptor superfamily, member 9 (4-1BB) antibody genes was constructed based on Western Reserve in our previous study. The present study evaluated the ability of this virus for cancer-targeted therapy both in vitro and in vivo. A complete morphological structure of Δ TK -Armed-VACV was verified using transmission electron microscopy. The antibody was co-expressed with the replication of Δ TK -Armed-VACV in vitro assessed by Western blot analysis, enzyme-linked immunosorbent assay, and quantitative real-time polymerase chain reaction. The 3-(4,5-dimethylthiazol-2-yl)-5-(3-rboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium, inner salt assay showed that the Δ TK -Armed-VACV exhibited significant tumor-specific cytotoxicity in vitro. The ΔTK-Armed-VACV inhibited the tumor growth in a 4T1 or A549 tumor-bearing mouse model. ELISpot assay showed that ΔTK-Armed-VACV-treated mice induced the expression of interferon-gamma, and lactate dehydrogenase–dependent cytotoxicity assay revealed that the Δ TK -Armed-VACV treatment activated tumor-specific cytotoxic T lymphocytes. The results indicated that oncolytic VACV with Western Reserve –mediated anti-human-PD-1 and anti-human-4-1BB antibody co-expression exerted a significant antitumor effect, indicating that the combination of oncolytic virotherapy and immunotherapy by the oncolytic VACV expressing one or more immune checkpoint genes might have satisfactory clinical expectations. • TK deleted oncolytic vaccinia virus co-expressed functional anti-PD-1 & anti-4-1BB • Anti-PD-1 & anti-4-1BB armed oncolytic vaccinia virus enhanced antitumor efficiency • Combined the immunotherapy with oncolytic virotherapy by armed oncolytic virus [ABSTRACT FROM AUTHOR]

To develop a drug-eluting polymer film which can be easily personalized and rapidly made on the electrode array of a cochlear implant during surgery. A precursor solution was prepared with poly lactic- co -glycolic acid (PLGA) and trichloromethane. Using a dip-coating method, the silicone electrode array (HiFocus 1J, Advanced Bionics) was coated in polymer film produced from the precursor solution containing one of three drugs: dexamethasone sodium phosphate (DSP), cytosine arabinoside hydrochloride (Ara-C), or nicotinamide adenine dinucleotide (NAD), and the release of these drugs from the polymer film was studied. The drug-eluting film on the electrode array was analyzed by environmental scanning electron microscopy (ESEM). The water contact angle and the impedance of the electrode array were measured before and after coating. Drug release kinetics was evaluated in a quasi-stationary release model, using high performance liquid chromatography every 24 h for 15 days. Five electrode arrays were tested with each of the three drugs in the polymer film coating. Before and after coating, ESEM studies revealed that the drug-loaded PLGA coating yielded a smooth covering with an average thickness of 1.02 ± 0.05 μm. The mass of the coated electrode increased by 1.00 ± 0.03 mg. The water contact angle decreased after coating (102 ± 0.6° vs 77 ± 1.6°, p < 0.01) but there was no significant change in the average impedance of the electrodes after coating (0.9 ± 0.22 kΩ vs 1.0 ± 0.18 kΩ, p > 0.05). An in vitro drug kinetics study revealed a faster release in the first 24 h (63.4 ± 0.6%) and a sustained release over the following 15 days (78.3 ± 1.7% in 2 days, 95.6 ± 1.0% in 7 days and 99.1 ± 0.4% in 14 days). The release rate was not affected by the drug, dose or the thickness of the coating. The dip-coating method is feasible for rapid casting of a drug-eluting PLGA film on an electrode array during CI surgery. The coated electrode array maintained its original morphology and became more hydrophilic. The loaded drug is released in a sustained manner and is easily regulated, and so the method might represent a potential application for clinical use in cochlear implantation. • Cochlear protective drugs are delivered via a coating on electrode array. • Drugs are loaded into degradable biocompatible PLGA film for coating. • Drug coating can be rapidly prepared on electrode array during surgery. • Drugs in coating can be slowly released in 15 days. • Drug concentration for releasing can be easily tailored during its preparation. [ABSTRACT FROM AUTHOR]