Immunotherapeutic potential of blinatumomab-secreting γ9δ2 T Cells.

Previous studies have explored the use of engineered blinatumomab-secreting autologous αß T cells for CD19-targeted cancer therapy. To create a more flexible allogeneic delivery system, we utilized γ9δ2 T cells rather than αß T cells in a similar application. First, we showed that γ9δ2 T cells could serve as effector cells for blinatumomab, and these effector memory cells could survive for at least 7 days after infusion. The genetically modified blinatumomab-secreting γ9δ2 T cells induced significant cytotoxicity in CD19+ tumor cell lines and primary cells from chronic lymphocytic leukemia patients. Of note, blinatumomab-secreting γ9δ2 T cells might also exhibit dual-targeting of CD19 and isopentenyl pyrophosphate, a universal tumor-associated antigen. Furthermore, blinatumomab-secreting γ9δ2 T cells killed CD19-transfected adherent cells, suggesting that the γ9δ2 T cells might be effective for treating solid tumors with appropriate cancer antigens. Together, these results demonstrate the promise of blinatumomab-secreting γ9δ2 T cells as a cancer therapy.

Supercritical Fluid-Assisted Fabrication of Pd Nanoparticles/Graphene Using a Choline Chloride-Oxalic Acid Deep Eutectic Solvent for Enhancing the Electrochemical Oxidation of Glycerol.

A green method for synthesizing Pd nanoparticles/graphene composites from a choline chloride-oxalic acid deep eutectic solvent (DES) without a reducing agent or a surfactant is reported. Deep eutectic solvents are usually composed of halide salts and hydrogen-bond donors, and many are biocompatible and biodegradable. The merits of deep eutectic solvents include that they serve as reducing agents and dispersants, and Pd nanoparticles are tightly anchored to graphene. The size and dispersion of Pd particles are improved when supercritical carbon dioxide (scCO2) is used because it has gaslike diffusivity and near-zero surface tension, which results in excellent wettability between the scCO2 and the carbon surface. The prepared sc-Pd NPs/GR/SPCE shows excellent activity toward glycerol oxidation compared to composites not fabricated by scCO2 processes. This study demonstrates the potential of using this scCO2-assisted protocol combined with deep eutectic solvents to further construct nanoparticles/graphene composites.

The potential of adoptive transfer of γ9δ2 T cells to enhance blinatumomab's antitumor activity against B-cell malignancy.

Blinatumomab, a bispecific T cell engager (BiTE) antibody targeting CD19 and CD3ε, can redirect T cells toward CD19-positive tumor cells and has been approved to treat relapsed/refractory B-cell acute lymphoblastic leukemia (R/R B-ALL). However, chemotherapeutic regimens can severely reduce T cells' number and cytotoxic function, leading to an inadequate response to blinatumomab treatment in patients. In addition, it was reported that a substantial portion of R/R B-ALL patients failing blinatumomab treatment had the extramedullary disease, indicating the poor ability of blinatumomab in treating extramedullary disease. In this study, we investigated whether the adoptive transfer of ex vivo expanded γ9δ2 T cells could act as the effector of blinatumomab to enhance blinatumomab's antitumor activity against B-cell malignancies in vivo. Repeated infusion of blinatumomab and human γ9δ2 T cells led to more prolonged survival than that of blinatumomab or human γ9δ2 T cells alone in the mice xenografted with Raji cells. Furthermore, adoptive transfer of γ9δ2 T cells reduced tumor mass outside the bone marrow, indicating the potential of γ9δ2 T cells to eradicate the extramedullary disease. Our results suggest that the addition of γ9δ2 T cells to the blinatumomab treatment regimens could be an effective approach to enhancing blinatumomab's therapeutic efficacy. The concept of this strategy may also be applied to other antigen-specific BiTE therapies for other malignancies.

[Optimization Method and Case Study of Air Pollution Emission Spatial Pattern].

Few of the current methods of improving air quality, including end-pipe treatment, industrial, energy and transportation structure adjustments, are from the viewpoint of the spatial pattern optimization of pollutant emissions. Therefore, based on factors such as natural environment, human health, pollutant transmission capability, and meteorological diffusion conditions, our research group used the threshold approach, natural breaks, spatial erasure, and other methods to define the layout area suitable for atmospheric pollution sources. Based on these results, the emissions pattern was optimized to achieve air quality improvement. Taking Guangdong Province as an example, we examined the application of the emissions pattern optimization of air quality improvement and atmospheric environment zoning. The results indicate that the first class area of environmental air quality accounts for 9% of total province area, the densely populated area accounts for 3%, the sensitive area of the national air quality monitor stations accounts for 15%, the pollutant accumulation area accounts for 22%, and the layout area suitable for atmospheric pollution sources primarily distributed in the west part of the province accounts for 60%. By shifting the non-thermal power industrial sources into those area, the concentration level of PM2.5 will decrease by 4% at the provincial scale and 10% at the city scale. Emissions pattern optimization has become an innovative aided support technology for the continuous improvement of air quality. In practical applications, it can be combined with energy and industrial structure adjustments, pollution control technology enhancements, and cross-regional prevention and control to formulate the most feasible air quality improvement plan.

[Source Apportionment of Air Pollutants for a Typical Pollution Event in Zhaoqing].

Based on observational data for pollutants and meteorology, this study analyzed the pollution episode that occurred during Dec 17th to 23th in 2018 in Zhaoqing, Guangdong Province, China. Using the source apportionment model CMAQ-ISAM and the hybrid receptor model, the regional contributions to air pollution were examined. The results showed that low-pressure conditions had an adverse effect on the diffusion of pollutants during this pollution episode in Zhaoqing. Prior to the pollution episode, pollutants were mainly derived from Zhaoqing and Qingyuan, accounting for 19.2% and 10.7% of pollutants, respectively. As well as pollutants from Guangdong Province, long-distance transport of pollutants from Jiangxi, Hunan, Hubei, and Shaanxi accounted for approximately 64.5% of the total during the non-pollution period. During the polluted episode, major cities in Pearl River Delta and the eastern part of Guangdong Province contributed more pollutants as a surface high-pressure field moved southward. Zhaoqing, Foshan, Dongguan, Guangzhou, and Huizhou contributed 25.5%, 14.8%, 9.8%, 9.5%, and 5.3% of the pollutants, respectively. Cities in the eastern part of Guangdong Province including Heyuan, Meizhou, Shanwei, Jieyang, Shantou, and Chaozhou contributed 13.7% of the total pollutants. In addition, pollutants from Fujian, Jiangxi, and the Yangtze River Delta accounted for approximately 32.9%. Furthermore, pollutants transported under marine influences were one of the main causes of this pollution episode in Zhaoqing.

[Emission Inventory of Atmospheric Pollutants and VOC Species from Crop Residue Burning in Guangdong Province].

An emission inventory of atmospheric pollutants from crop residue burning in Guangdong for the period 2008-2016 was developed, based on crop yield data. Emissions of species of volatile organic compounds(VOCs)and corresponding ozone formation potential (OFP) in 2016 were also estimated. Results showed that emissions of atmospheric pollutants from crop residue burning in 2013-2016 were lower than in 2008-2012. This was mainly due to the policy of prohibiting open burning of straw and to improvement of rural living standards, which reduced the proportion of straw burning. In 2016, emissions of SO2, NOx, NH3, CH4, EC, OC, NMVOC, CO, and PM2.5 were 2443.7, 16187.9, 6943.8, 29174.4, 3625.5, 14830.7, 65612.6, 591613.9, and 49463.0 t, respectively. Rice straw burning was the main source of pollutants, accounting for about 68.55% of total pollutant emissions. The five municipalities with highest atmospheric pollutant emissions were Zhanjiang, Maoming, Meizhou, Zhaoqing, and Shaoguan, together accounting for about 58.63% of total emissions. The top 10 VOC species for mass-based emissions consisted of ethylene, acetaldehyde, formaldehyde, benzene, ethyne, propylene, ethane, toluene, propane, and propionaldehyde, together contributing 67.91% to total emissions. The top ten OFP-based VOC species were ethylene, formaldehyde, acetaldehyde, propylene, 1-butylene, propionaldehyde, toluene, acrolein, isoprene, and crotonaldehyde, accounting for 80.83% of total OFP.

[Air Quality Subarea Management:A Case Study of Guangdong Province].

To meet the requirements of regional air quality management (AQM), the Air Quality Subarea Management (AQSM) system was proposed. A case study was conducted for Guangdong Province. By using the method of air quality numerical simulation and satellite remote sensing inversion analysis, the key factors were selected from the meteorological simulation field, the pollutant concentration simulation field, and the satellite image interpretation to form the index system for AQSM. On this basis, a hierarchical cluster analysis method was used to divide Guangdong Province into three types of AQSM:Strict Control Subarea, Continuous Improvement Subarea, and Coordinated Development Subarea. It was shown that the Strict Control Subarea, Continuous Improvement Subarea, and Coordinated Development Subarea in Guangdong Province covered 16.3%, 28.0%, and 55.7%, respectively. The Strict Control Subarea in the Pearl River Delta, Eastern Guangdong, Western Guangdong, and Northern Guangdong accounted for 27.9%, 19.3%, 4.4%, and 12.5%, respectively, and the subarea should implement the most stringent AQM policies to promote air quality improvement. The Continuous Improvement Subarea in the Pearl River Delta, Eastern Guangdong, Western Guangdong, and Northern Guangdong accounted for 34.4%, 15.8%, 7.8%, and 34.5%, respectively, and the subarea should implement relatively strict AQM policies to ensure sustained and stable standards. The Coordinated Development Subarea in the Pearl River Delta, Eastern Guangdong, Western Guangdong, and Northern Guangdong accounted for 37.7%, 64.9%, 87.8%, and 53.0%, respectively, and the subarea could implement more liberal AQM policies to ensure relatively good air quality. In general, the strict AQM policies in Guangdong Province should be mainly concentrated in the Pearl River Delta region, followed by Northern Guangdong, Eastern Guangdong, and Western Guangdong in order.

[Capacity Simulation Method Based on Regional Transfer Matrix and PM2.5 Concentration Target Constraint].

Atmospheric environmental capacity is an important reference in environmental planning. To meet the PM2.5 standard, a new method is proposed to balance the capacity among cities of Guangdong, with screening of the most unfavorable meteorological year and combining it with the regional transportation calculated by the CAMx-PSAT module. Pollutant overloading and capacity scenarios were also calculated. The results showed that, under the constraints of the cities' annual PM2.5 ≤ 35 µg·m-3, the capacities of SO2, NOx, NH3, and PM2.5 in Guangdong were about 6.8×105 tons, 1.35×106 tons, 4.6×105 tons, and 5.1×105 tons, respectively. Based on the benchmark scenario, SO2 emissions in Guangdong were overloaded by 10%, and the emissions of NOx, NH3, and PM2.5 exceeded by 12%, 9%, and 20%, respectively, compared to those of the capacity scenario. Ranked by the number of overloaded species in Guangdong, the cities of Guangzhou, Foshan, Zhongshan, and Qingyuan were on top. When achieving the capacity scenario, the annual PM2.5 concentration in Guangdong was about 30 µg·m-3, which meets the national secondary ambient air quality standard.

Evidence of preserved collagen in an Early Jurassic sauropodomorph dinosaur revealed by synchrotron FTIR microspectroscopy.

Fossilized organic remains are important sources of information because they provide a unique form of biological and evolutionary information, and have the long-term potential for genomic explorations. Here we report evidence of protein preservation in a terrestrial vertebrate found inside the vascular canals of a rib of a 195-million-year-old sauropodomorph dinosaur, where blood vessels and nerves would normally have been present in the living organism. The in situ synchrotron radiation-based Fourier transform infrared (SR-FTIR) spectra exhibit the characteristic infrared absorption bands for amide A and B, amide I, II and III of collagen. Aggregated haematite particles (α-Fe2O3) about 6∼8 µm in diameter are also identified inside the vascular canals using confocal Raman microscopy, where the organic remains were preserved. We propose that these particles likely had a crucial role in the preservation of the proteins, and may be remnants partially contributed from haemoglobin and other iron-rich proteins from the original blood.

Human acellular cartilage matrix powders as a biological scaffold for cartilage tissue engineering with synovium-derived mesenchymal stem cells.

In our previous study, we found that cartilage fragments from osteoarthritic knee promoted chondrogenesis of mesenchymal stem cells. In this study, we further transformed the cartilage tissues into acellular cartilage matrix (ACM) and explored the feasibility of using ACM as a biological scaffold. Nonworn parts of cartilage tissues were obtained during total knee arthroplasty (TKA) surgery and were successfully fabricated into ACM powders. The ACM powders and human synovium-derived mesenchymal stem cells (SMSCs) were mixed into collagen gel for in vitro culture. Histological results showed a synergistic effect of ACM powders and chondrogenic growth factors in the formation of engineered cartilage. The findings of real-time polymerase chain reaction (PCR) suggested that ACM powders had the potential of promoting type II collagen gene expression in the growth factors-absent environment. Moreover, with growth factors induction, the ACM powders could reduce the hypertrophy in chondrogenesis of SMSCs. In summary, ACM powders could serve as a functional scaffold that benefited the chondrogenesis of SMSCs for cartilage tissue engineering.

Cartilage fragments from osteoarthritic knee promote chondrogenesis of mesenchymal stem cells without exogenous growth factor induction.

Extracellular matrix (ECM) is thought to participate significantly in guiding the differentiation process of mesenchymal stem cells (MSCs). In this study, we hypothesized that cartilage fragments from osteoarthritic knee could promote chondrogenesis of MSCs. Nonworn parts of cartilage tissues were obtained during total knee arthroplasty (TKA) surgery. Cartilage fragments and MSCs were wrapped into fibrin glue; and the constructs were implanted subcutaneously into nude mice. Histological analysis showed neocartilage-like structure with positive Alcian blue staining in the cartilage fragment-fibrin-MSC constructs. However, constructs with only MSCs in fibrin showed condensed appearance like MSCs in the pellet culture. Gene expression of type II collagen in the constructs with 60 mg cartilage fragments were significantly elevated after 4 weeks of implantation. Conversely, the constructs without cartilage fragments failed to express type II collagen, which indicated MSCs did not differentiate into a chondrogenic lineage. In conclusion, we demonstrated the effect of cartilage fragments from osteoarthritic knee in promoting chondrogenic differentiation of MSCs. This may be a favorable strategy for MSC chondrogenesis without exogenous growth factor induction.

Ex vivo expansion of dendritic-cell-activated antigen-specific CD4+ T cells with anti-CD3/CD28, interleukin-7, and interleukin-15: potential for adoptive T cell immunotherapy.

There is an increasing realization that the failure of adoptive therapy with cytotoxic T lymphocytes in the autologous setting, at least in part, results from the lack of help from antigen-specific CD4+ T cells. To incorporate these cells into this treatment strategy, it is not known whether currently used ex vivo culture conditions are adequate for expanding and charting these T cells with the desired qualities for optimal in vivo activity. In this study, we show that stimulation with agonistic antibodies to CD3 plus CD28 (anti-CD3/CD28), a commonly used method for CD4+ T cell expansion, is unable to expand dendritic-cell-activated hepatitis B virus (HBV)-specific CD4+ T cells to clinical relevant numbers. Whereas, in combination with interleukin(IL)-7 and IL-15, it leads to a 4000-fold expansion of HBV-specific CD4+ T cells in 2 weeks. This outcome is correlated with the anti-apoptosis effect of IL-7 and IL-15. Importantly, antigen specificity is preserved during expansion. Although a late addition of IL-2 to the anti-CD3/CD28-expanding cultures also results in robust expansion, this expansion condition renders HBV-specific CD4+ T cells more sensitive to cytokine withdrawal-, activation-, and transforming growth factor-beta-induced cell death compared to those expanded in IL-7 and IL-15. Moreover, NKG2D rather than 4-1BB, whose ligands are constitutively expressed on tumor cells, is significantly up-regulated on IL-7/IL-15-expanded HBV-specific CD4+ T cells, and its engagement promotes expansion and interferon-gamma production by these cells and thus may serve to provide co-stimulation to T cells once they reach tumor tissues. Collectively, these results may have important therapeutic implications for adoptive T cell therapy.

Transforming growth factor-beta induces apoptosis in antigen-specific CD4+ T cells prepared for adoptive immunotherapy.

Transforming growth factor-beta (TGF-beta), found at the site of most tumors, has been recognized as one of the mechanisms involved in tumor immunological escape. To evaluate its impact on adoptive immunotherapy against cancer, we examined the susceptibility of tumor-specific T cells to TGF-beta in the setting of these T cells being prepared for adoptive transfer. Hepatitis B virus (HBV)-specific CD4(+) T cells were ex vivo generated by activating with HBV-transfected dendritic cells and selecting with antibodies to CD25 activation molecules, and then expanded with antibodies to CD3/CD28. These T cells expressed higher levels of the type II TGF-beta receptor than nai;ve T cells and exhibited enhanced apoptosis when exposed to TGF-beta. The underlying apoptotic pathway was linked to the dissipation of the mitochondrial inner membrane potential and activation of caspase-9. The absence of caspase-8 activity in TGF-beta-treated T cells suggests that the death receptor system may not be involved in this type of apoptosis. Interleukin-2 (IL-2), which is concomitantly administered with tumor-specific T cells in adoptive immunotherapy, was unable to protect HBV-specific CD4(+) T cells from the pro-apoptotic effect of TGF-beta when added simultaneously with TGF-beta. Interesting, IL-2-pretreated T cells displayed the type II TGF-beta receptor at lower levels and were more resistant to TGF-beta. Together, our findings indicate that the effectiveness of adoptive cancer immunotherapy may be impaired by tumor-derived TGF-beta and appropriate manipulation of exogenous IL-2 might overcome this hurdle.