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Ex vivo-expanded, allogeneic natural killer (NK) cells can be used for the treatment of various types of cancer. In allogeneic NK cell therapy, NK cells from healthy donors must be expanded in order to obtain a sufficient number of highly purified, activated NK cells. In the present study, we established a simplified and efficient method for the large-scale expansion and activation of NK cells from healthy donors under good manufacturing practice (GMP) conditions. After a single step of magnetic depletion of CD3(+) T cells, the depleted peripheral blood mononuclear cells (PBMCs) were stimulated and expanded with irradiated autologous PBMCs in the presence of OKT3 and IL-2 for 14 days, resulting in a highly pure population of CD3(-)CD16(+)CD56(+) NK cells which is desired for allogeneic purpose. Compared with freshly isolated NK cells, these expanded NK cells showed robust cytokine production and potent cytolytic activity against various cancer cell lines. Of note, expanded NK cells selectively killed cancer cells without demonstrating cytotoxicity against allogeneic non-tumor cells in coculture assays. The anti-tumor activity of expanded human NK cells was examined in SCID mice injected with human lymphoma cells. In this model, expanded NK cells efficiently controlled lymphoma progression. In conclusion, allogeneic NK cells were efficiently expanded in a GMP-compliant facility and demonstrated potent anti-tumor activity both in vitro and in vivo.

IntroductionAllodynia, which can be induced by paclitaxel administration, is the presence of pain as a result of a stimulus that does not usually provoke pain. Many studies have investigated the analgesic efficacy of acupuncture, including laser acupuncture (LA) and electroacupuncture (EA). Although pain-related diseases are relatively common, few studies have analyzed the analgesic effects and mechanisms of LA combined with EA. The purpose of this study was to investigate the therapeutic effect and mechanism of manual acupuncture (MA), EA, LA, and combined therapy (LA + EA) in a paclitaxel-induced allodynia rat model.MethodsA total of 56 rats were classified into eight groups: a normal (Nor, n = 7), a control (Con, n = 7), an MA (n = 7), an EA (n = 7), a 650-nm LA (650LA, n = 7), an 830-nm LA (830LA, n = 7), a 650-nm LA combined with EA (650LA + EA, n = 7), and an 830-nm LA combined with EA group (830LA + EA, n = 7). Allodynia was induced by intraperitoneal injection of 2 mg/kg of paclitaxel every other day for a total of four times except the Nor group. Acupuncture treatments were conducted at the points of Jungwan (CV12) and Joksamni (ST36) once every other day for 6 min, for a total of nine times. Withdrawal response reaction times and force intensity of the foot were measured before the start of the experiment, after the 4th paclitaxel administration (day 8), and after the 9th and last treatment (day 15). On the 16th day, mRNA and protein expression in the spinal nerves was assessed, and a metabolome analysis of the animals’ feces was performed.Results and discussionOur analyses show that 650LA + EA treatment resulted in an upregulation of protein expression related to pain relief and nerve regeneration, whereas 830LA + EA treatment led to significant changes in metabolomes. This study demonstrates that a combination treatment of EA and LA can suppress allodynia and promote upregulation of protein expression related to nerve regeneration and is effective in changing the intestinal microbiome. Further large-scale research is required to assess the exact mechanism underlying the therapeutic effect of this combination treatment in pain-related diseases.

Amorphous biodegradable copolyesters have been synthesized using various alicyclic monomers, including 1,4-cyclohexanedimethanol (CHDM), 1,4-cyclohexanediol (CHDO), isosorbide (1,4:3,6-dianhidro-D-sorbitol) (IS), and 1,4-cyclohexane dicaboxylic acid (CHDA), to enhance the thermal properties of polyesters. We have studied the effect of alicyclic monomers on the thermal properties, especially, on the glass transition temperature (Tg) of the amorphous copolyesters. On incorporating alicyclic diol monomers into poly(ethylene succinate-co-terephthalate) (PEST), Tg of amorphous copolyesters was greatly increased. As the rigidity of alicyclic monomers increases, the effect becomes more significant. By comparing copolyesters synthesized with the same molar ratio of alicyclic monomers in the feed, we found that the molar ratio of IS in the copolyester product was much smaller than those of CHDM and CHDO. Nonetheless, IS induced the largest Tg enhancement in copolyesters even though a much smaller molar ratio of IS was incorporated in the product compared to CHDM and CHDO. To study the effects of alicyclic diacid monomers, copolyesters containing CHDA were synthesized. As in the case of alicyclic diols, CHDA was found to be very effective in enhancing the thermal properties. Among these copolyesters produced, PEI30ST synthesized with 30 mol% of succinic acid (SA) and 70 mol% of dimethyl terephthalate (DMT) as diacid and 30 mol% of IS as diol exhibited the highest Tg of 74.2 oC. And poly(ethylene-co-CHDM succinate-co-terephthalate) (PEC30ST70) and poly(ethylene-co-IS succinate-co-terephthalate) (PEI30ST70) exhibited high optical transmittance and were confirmed to be biodegradable, exhibiting about 50% and 40% biodegradability, respectively, after 50 days in a biodegradability test under compost conditions. Hence, these novel transparent biodegradable polyesters could be highly useful in flexible packaging applications requiring transparency. [ABSTRACT FROM AUTHOR]

Background: Glioblastoma multiforme (GBM) is characterized by extensive local invasion, which is in contrast with extremely rare systemic metastasis of GBM. Molecular mechanisms inhibiting systemic metastasis of GBM would be a novel therapeutic candidate for GBM in the brain.Methods: Patient-derived GBM cells were primarily cultured from surgical samples of GBM patients and were inoculated into the brains of immune deficient BALB/c-nude or NOD-SCID IL2Rgamma(null) (NSG) mice. Human NK cells were isolated from peripheral blood mononucleated cells and expanded in vitro.Results: Patient-derived GBM cells in the brains of NSG mice unexpectedly induced spontaneous lung metastasis although no metastasis was detected in BALB/c-nude mice. Based on the difference of the innate immunity between two mouse strains, NK cell activities of orthotopic GBM xenograft models based on BALB/c-nude mice were inhibited. NK cell inactivation induced spontaneous lung metastasis of GBM cells, which indicated that NK cells inhibit the systemic metastasis. In vitro cytotoxic activities of human NK cells against GBM cells indicated that cytotoxic activity of NK cells against GBM cells prevents systemic metastasis of GBM and that NK cells could be effective cell therapeutics against GBM. Accordingly, NK cells transplanted into orthotopic GBM xenograft models intravenously or intratumorally induced apoptosis of GBM cells in the brain and showed significant therapeutic effects.Conclusions: Our results suggest that innate NK immunity is responsible for rare systemic metastasis of GBM and that sufficient supplementation of NK cells could be a promising immunotherapeutic strategy for GBM in the brain. [ABSTRACT FROM AUTHOR]