The petroleum ether fraction of ethanol extracts from Boenninghausenia sessilicarpawas isolated by combination of several chromatographic methods including silica gel, ODS, and Sephadex LH-20 column chromatography, and finally purified by preparative HPLC. The structures of the isolated compounds were identified based on the spectral data. As a result, 15 coumarin compounds were isolated and identified from the petroleum ether extraction. Their structures were determined as osthenon (1), murrangatin (2), 3-(1,1-dimethylallyl)-8-hydroxy-7-methoxycoumarin (3), xanthotoxin (4), isopimpinellin (5), chalepensin (6), isodemethylfuropinarine (7), imperatorin (8), phellopterin (9), heraclenol (10), byakangelicin (11), neobyakangelicol (12), chalepin (13), luvangetin (14), 3-(1, 1-dimethylallyl)-xanthyletin (15). Among them, compounds 1 - 3, 6 - 10 and 14 -15 were firstly isolated from B. sessilicarpa.

The generation of T cells with maximal anti-tumor activities will significantly impact the field of T-cell-based adoptive immunotherapy. In this report, we found that OKT3/IL-2-stimulated T cells were phenotypically more heterogeneous, with enhanced anti-tumor activity in vitro and when locally administered in a solid tumor mouse model. To further improve the OKT3/IL-2-based T cell manufacturing procedure, we developed a novel T cell stimulation and expansion method in which peripheral blood mononuclear cells were electroporated with mRNA encoding a chimeric membrane protein consisting of a single-chain variable fragment against CD3 and the intracellular domains of CD28 and 4-1BB (OKT3-28BB). The expanded T cells were phenotypically and functionally similar to T cells expanded by OKT3/IL-2. Moreover, co-electroporation of CD86 and 4-1BBL could further change the phenotype and enhance the in vivo anti-tumor activity. Although T cells expanded by the co-electroporation of OKT3-28BB with CD86 and 4-1BBL showed an increased central memory phenotype, the T cells still maintained tumor lytic activities as potent as those of OKT3/IL-2 or OKT3-28BB-stimulated T cells. In different tumor mouse models, T cells expanded by OKT3-28BB RNA electroporation showed anti-tumor activities superior to those of OKT3/IL-2 T cells. Hence, T cells with both a less differentiated phenotype and potent tumor killing ability can be generated by RNA electroporation, and this T cell manufacturing procedure can be further optimized by simply co-delivering other splices of RNA, thus providing a simple and cost-effective method for generating high-quality T cells for adoptive immunotherapy.
Animals ; CD28 Antigens ; genetics ; immunology ; Electroporation ; Humans ; Immunity, Cellular ; Interleukin-2 ; immunology ; K562 Cells ; Mice ; Muromonab-CD3 ; immunology ; Neoplasms, Experimental ; genetics ; immunology ; pathology ; RNA, Messenger ; genetics ; immunology ; T-Lymphocytes ; immunology ; Tumor Necrosis Factor Receptor Superfamily, Member 9 ; genetics ; immunology