T cell receptor gene-modified allogeneic T cells with siRNA for endogenous T cell receptor induce efficient tumor regression without graft-versus-host disease.

Adoptive immunotherapy using genetically engineered patient-derived lymphocytes to express tumor-reactive receptors is a promising treatment for malignancy. However, utilization of autologous T cells in this therapy limits the quality of gene-engineered T cells, thereby inhibiting the timely infusion of the cells into patients. In this study, we evaluated the anti-tumor efficacy and the potential to induce graft-versus-host disease (GVHD) in T cell receptor (TCR) gene-engineered allogeneic T cells that downregulate the endogenous TCR and HLA class I molecules with the aim of developing an "off-the-shelf" cell product with expanded application of genetically engineered T cells. We transduced human lymphocytes with a high-affinity TCR specific to the cancer/testis antigen NY-ESO-1 using a novel retrovirus vector with siRNAs specific to the endogenous TCR (siTCR vector). These T cells showed reduced expression of endogenous TCR and minimized reactivity to allogeneic cells in vitro. In non-obese diabetic/SCID/γcnull mice, TCR gene-transduced T cells induced tumor regression without development of GVHD. A lentivirus-based CRISPR/Cas9 system targeting ß-2 microglobulin in TCR gene-modified T cells silenced the HLA class I expression and prevented allogeneic CD8+ T cell stimulation without disrupting their anti-tumor capacity. This report is the first demonstration that siTCR technology is effective in preventing GVHD. Adoptive cell therapy with allogeneic T cells engineered with siTCR vector may be useful in developing an "off-the-shelf" therapy for patients with malignancy.

An investigation of browsing enrichment, especially non-leaf foraging, on giraffes (Giraffa camelopardalis reticulata) at Kyoto City Zoo in Japan.

Browsing enrichment may aid in developing species-specific behaviors for giraffes managed in zoos as a means of improving animal welfare. By nature, giraffes are tree-feeding animals, including tree bark, but the extent of food other than leaves as a form of browsing enrichment has not been well investigated. Therefore, to investigate the effectiveness of non-leaf foraging, three giraffes at the Kyoto City Zoo in Japan were observed for 228 h from May 2019 to February 2020. In conjunction with behavioral instantaneous sampling, tree use (landscape tree or enrichment branch) and plant part (leaves, twigs, or barks) were recorded by the 1-0 sampling method. There was no significant change in the foraging behavior on the leaves of enriched branches, nor was there any significant change in the foraging behavior of the giraffes, except for one animal in the deciduous phase. No significant changes were observed in rumination or other behaviors between the two phases. Although vegetation foraging behavior significantly decreased, except for one animal, dry hay foraging behavior significantly increased in all the animals during the deciduous phase. Some individuals also showed a significant increase in the foraging behavior for non-leafy parts of the enrichment branches (twigs and bark) during the deciduous phase. This suggests that in some tree species, giraffes forage on the bark and twigs to compensate for the loss of leaves during the deciduous phase, similar to feeding on hay or hay cubes as a substitute for tree leaves.

Effects of browsing enrichment associated with the temperature-humidity index and landscaping trees in giraffes (Giraffa camelopardalisreticulata).

Enclosure environments for captive giraffes can be improved by promoting species-specific behaviors and extending foraging behavior. To date, however, the effects of climatic (temperature-humidity index, THI) and environmental factors (landscaping trees) on the enrichment of captive settings have not been studied. Therefore, the present study explored the effects of browsing enrichment on the licking behavior of captive giraffes. From May 2019 to February 2020, three giraffes in the Kyoto City Zoo, Japan, were observed for 270 h over two consecutive seasons (pre- and post-deciduous). Overall, branch foraging behavior and licking behavior were weakly and negatively correlated. In the pre-deciduous period, THI was significantly and negatively correlated with giraffe activity, and some individuals exhibited significantly reduced foraging behavior. Therefore, browsing enrichment under extreme heat may not improve giraffe rearing environments. Moreover, in the post-deciduous period, with decreased availability of leaves on landscaping trees, the non-branch foraging behavior of giraffes was significantly increased, with a corresponding significant increase in licking behavior. Therefore, landscaping trees affect the foraging behavior of captive giraffes.

Chemical characterization of the milk oligosaccharides of some Artiodactyla species including giraffe (Giraffa camelopardalis), sitatunga (Tragelaphus spekii), deer (Cervus nippon yesoensis) and water buffalo (Bubalus bubalis).

Mammalian milk/colostrum usually contains oligosaccharides along with the predominant disaccharide lactose. It has been found that the number and identity of these milk oligosaccharides varies among mammalian species. Oligosaccharides predominate over lactose in the milk/colostrum of Arctoidea species (Carnivora), whereas lactose predominates over milk oligosaccharides in Artiodactyla including cow, sheep, goat, camel, reindeer and pig. To clarify whether heterogeneity of a variety of milk oligosaccharides is found within other species of Artiodactyla, they were studied in the milk of giraffe, sitatunga, deer and water buffalo. The following oligosaccharides were found: Neu5Ac(α2-3)[GalNAc(ß1-4)]Gal(ß1-4)Glc (GM2 tetrasaccharide), and Gal(α1-3)Gal(ß1-4)Glc (isoglobotriose) in giraffe milk; Neu5Ac(α2-3)Gal(ß1-4)Glc (3'-SL), Neu5Ac(α2-6)Gal(ß1-4)Glc (6'-SL), Gal(α1-4)Gal(ß1-4)Glc (globotriose) and isoglobotriose in sitatunga colostrum; Gal(ß1-3)Gal(ß1-4)Glc (3'-GL), Gal(ß1-6)Gal(ß1-4)Glc (6'-GL), isoglobotriose, Gal(ß1-4)GlcNAc(ß1-3)Gal(ß1-4)Glc (lacto-N-neotetraose, LNnT), Gal(ß1-4)Glc-3'-O-SO3 (3'-O-lactose sulphate) in deer milk; 3'-GL, isoglobotriose and Gal(ß1-3)Gal(ß1-3)Gal(ß1-4)Glc (3',3″-digalactosyllactose, DGL) in water buffalo colostrum. Thus it was shown that the milk oligosaccharides are heterogeneous among these Artiodactyla species.

Temperature sensitivity of microbial respiration of fine root litter in a temperate broad-leaved forest.

The microbial decomposition respiration of plant litter generates a major CO2 efflux from terrestrial ecosystems that plays a critical role in the regulation of carbon cycling on regional and global scales. However, the respiration from root litter decomposition and its sensitivity to temperature changes are unclear in current models of carbon turnover in forest soils. Thus, we examined seasonal changes in the temperature sensitivity and decomposition rates of fine root litter of two diameter classes (0-0.5 and 0.5-2.0 mm) of Quercus serrata and Ilex pedunculosa in a deciduous broad-leaved forest. During the study period, fine root litter of both diameter classes and species decreased approximately exponentially over time. The Q10 values of microbial respiration rates of root litter for the two classes were 1.59-3.31 and 1.28-6.27 for Q. serrata and 1.36-6.31 and 1.65-5.86 for I. pedunculosa. A significant difference in Q10 was observed between the diameter classes, indicating that root diameter represents the initial substrate quality, which may determine the magnitude of Q10 value of microbial respiration. Changes in these Q10 values were related to seasonal soil temperature patterns; the values were higher in winter than in summer. Moreover, seasonal variations in Q10 were larger during the 2-year decomposition period than the 1-year period. These results showed that the Q10 values of fine root litter of 0-0.5 and 0.5-2.0 mm have been shown to increase with lower temperatures and with the higher recalcitrance pool of the decomposed substrate during 2 years of decomposition. Thus, the temperature sensitivity of microbial respiration in root litter showed distinct patterns according to the decay period and season because of the temperature acclimation and adaptation of the microbial decomposer communities in root litter.