A dendritic/tumor fusion cell vaccine enhances efficacy of nanobody-based CAR-T cells against solid tumor.

Background: Chimeric antigen receptor (CAR) T-cell therapy is practical in treating cancers of hematopoietic origin, but of that in solid tumors compromises efficacy for the loss of the antigen recognized by the CAR. However, dendritic cell (DC)/tumor fusion vaccines present a spectrum of known or unknown tumor antigens to stimulate T cell expansion and enhanced T cell response. Developing a new strategy of enhanced nanobody-based CAR-T (Nb-CAR-T) cells antitumor activity by DC/tumor fusion vaccines stimulation would provide guidance for more effective CAR-T cell therapies. Methods: Considering the therapeutic potential of nanobody (Nb), we first screened EGFRvIII Nb, then constructed and verified the function of EGFRvIII Nb-CAR-T cells in vitro and in vivo. We further combined DC/tumor fusion vaccines to boost EGFRvIII Nb-CAR-T cells antitumor effect, which was evaluated in vitro Nb-CAR-T cell function and in the tumor-bearing xenograft mouse models. Results: We had for the first time successfully selected EGFRvIII Nb for the generation of the novel EGFRvIII Nb-CAR-T cells. Importantly, our results suggested that DC/tumor fusion vaccines stimulate Nb-CAR-T cells response not only in improving T cell proliferation, T cell activation, cytokine secretion and tumor-specific cytotoxicity in vitro, but also significantly reducing tumor burden, prolonging survival and improving Nb-CAR-T cells infiltration. Conclusions: We have innovatively shown that DC/tumor fusion vaccines significantly enhance the efficacy of Nb-CAR-T cells against solid tumors. This new strategy has provided a promising therapeutic platform for promoting the clinical treatment of CAR-T cells therapy.

[Influence of Antecedent Dry Days on Nitrogen Removal in Bioretention Systems].

Bioretention systems have become an optimal technology during the construction of the sponge city, but its nitrogen removal performance can be affected by antecedent dry days (ADD). This study was designed to investigate the effects of different lengths of ADD (1,2,3, 5, 7, 12, and 22 d) on nitrogen removal performance using a series of laboratory-scale bioretention systems to form seven constant alternate drying-rewetting regimes. The influence mechanism was further investigated by analyzing the spatial distribution of nitrogen reductase activity and microbial community structure under different drying-rewetting regimes. The results showed that the ammonium removal efficiency was not significantly affected by ADD, while exhibiting high variation depending on the hydraulic permeability of the filler and plant growth conditions. The nitrate and total nitrogen removal efficiency decreased as the length of ADD increased form 7 d to 22 d. In addition, the spatial distribution of nitrate reductase (NaR), nitrite reductase (NiR), and hydroxylamine reductase (HyR) were affected by ADD to some extent. It was found that the soil moisture of submerged layer (SL) regulated the nitrogen processes. The nitrate dissimilatory reduction to ammonium (DNRA) can occur in the SL through secondary catalytic reduction by nitrogen reductases, thus affecting the removal of ammonium. The soil microbial community structure and its spatial distribution could be altered by ADD significantly, and the removal of multiple nitrogen species was partly affected. Thereinto, under shorter ADD values of 1, 2, 3, and 5 d, the dominant phylum was Firmicutes, a group of denitrifying microbes, and its dominant genus, Clostridium_sensu_stricto_1, also had the function of DNRA. The results of the study confirmed that ADD has a certain effect on the nitrogen removal capacity and nitrogen reductase activity, while resulting in spatial changes in the microbial community structure in the bioretention system under constant drying-rewetting conditions.

Vitamin D levels correlate with lymphocyte subsets in elderly patients with age-related diseases.

Hypovitaminosis D is associated with age-related illnesses, including hypertension, cardiovascular disease (CRVD), cerebrovascular disease (CAD) and type 2 diabetes mellitus (T2DM). In our retrospective observational study, blood samples of elderly healthy controls (n = 461) and patients with age-related diseases (n = 8,621) were subjected to flow-cytometry in order to determine correlations between age-related diseases and cluster of differentiation 4 (CD4), CD8, CD3, and CD19 lymphocyte markers, as well as serum levels of 25-hydroxyvitamin D2 (25(OH)D2) and 25-hydroxyvitamin D3 (25(OH)D3). More than 70% of the patients in each disease group had total vitamin D < 20 ng/mL (P < 0.001). In CRVD patients, CD3 and CD19 correlated (P < 0.05) with 25(OH)D3. In CAD patients, CD8, CD4, CD19 and CD4/CD8 correlated (P < 0.05) with 25(OH)D2, and CD8 correlated (P < 0.05) with 25(OH)D3. In T2DM and hypertension patients, CD8, CD3, CD19 and CD4/CD8 correlated with 25(OH)D3. Progressive trends (P < 0.05) towards increased CD8 and CD4/CD8 were observed in vitamin-D-deficient T2DM and hypertension patients. Significant differences (P < 0.05) in CD8 were observed in vitamin-D-deficient CAD patients, whereas significant differences (P < 0.05) in CD8 and CD19 were observed in CRVD patients. Higher CD8 and CD4/CD8 in 25(OH)D-deficient T2DM and hypertension patients suggested a Th1 lymphocyte profile induction. Increases in CD8-positive lymphocytes suggested a similar, less pronounced effect in vitamin-D-deficient CRVD and CAD patients.