CAR Activation Research Articles

Analysis of Plant Seed Distribution Activities by Immigration Polytechnic Cadets in Car Free Day Activities in Tangerang


 
 The purpose of carrying out the activity of distributing plant seeds by the Immigration Polytechnic Cadets with the Class I Non TPI Tangerang Immigration Office in this car free day activity is a form of concern for the surrounding environment, especially the people of Tangerang City. The activity, which was carried out with the theme "Together to Know The Culture and Social Development of Community", was in the form of devotional work activities, distribution of plant seeds to the community, as well as the implementation of socialization related to immigration by the Class I Non TPI Tangerang Immigration Office and socialization related to Polytechnic. Immigration by Immigration Polytechnic Cadets. This activity is a form of social responsibility for the Immigration Polytechnic Cadets as new citizens of Tangerang City and their environment.
 
 

Towards Versatile Protein Fabrics: UV Protection and Antimicrobial Property Dyeing with Centella Asiatica (L.) into Silk

Centella asiatica (CA) is known as a plant with therapeutic effect called Gotu Kola. In this study, silk was dyed using undried raw CA (CA-R) and dried CA (CA-D) extracts; the samples were evaluated for properties such as dyeability; fastness to light, perspiration, and washing; antimicrobial properties; and ultraviolet (UV) protection. Before and after dyeing, the sugar content and turbidity were measured to determine whether dyeing was efficient. It was confirmed that the dyeability and functionality of the CA-D-dyed samples were better than those of CA-R-dyed samples. For the fastness test, the fastness to light, at 4 or higher, was very good; furthermore, all the samples were in good condition for washing and perspiration, with grades ranging from 4 to 5. The UV-B protection value of the sample was 75 % higher than that of the undyed silk. The antimicrobial test showed that, in comparison to the original silk, silk dyed in the extracts exhibited 99.9 % better results, for concentrations above 5 µg/ml. Up to a concentration of 25 µg/ml, no significant decrease in cell viability was noted, indicating that the antimicrobial activity of CA-R is excellent.

Non-verbal Communication and Touchless Activation of a Radio-Controlled Car via Facial Activity Recognition

Many smart glasses technologies are being developed to improve the working efficiency or quality of life in various fields. In some enterprises, these technologies are used to help improve the working quality and productivity and minimize data loss. In real life, smart glasses are applied as an entertainment device with augmented/virtual reality or as an assistive manipulator for the physically challenged. Thus, these technologies have mainly adopted various operating systems depending on usages, such as a touchpad, remote control, and voice recognition. However, conventional operating methods have limitations in non-verbal and noisy situations where people cannot use both hands. In this study, we present a method of detecting a facial signal for touchless activation using a transducer. We acquired a facial signal amplified by a lever mechanism using a load cell on the hinge of an eyewear. We then classified the signal and obtained their accuracy by calculating the confusion matrix with classified categories through a machine learning technique, i.e., the support vector machine. We can activate an actuator, such as a radio-controlled car, through a classified facial signal by using an eyewear-type signal transducer. Overall, our operating system can be useful for activating the actuator or transmitting a message through the classified facial activities in non-verbal situations and in situations where both hands cannot be used.

Smart Usage of Open Source License Plate Detection and Using IoT Tools for Private Garage and Parking Solutions

Open source license plate detection using internet of things is a modern day technology whereby the personal car activity entering the garage is identified by the cameras and the garage lights are lit up. When it leaves the garage lights are switched off. For an IP camera, the main image processing and search for number plates. The research here provides an important strategy for identifying, segmenting and understanding the characters within the located plate. We have proposed in this study a license plate detection algorithm and using internet of things simulation tool Node-red identifies the state transition for a car activity and light movements which are proposed in this work. The use of ‘Internet of things’ due to dynamic and self adapting nature of IoT to one server that facilitate in tracing a vehicle more easily over having manual database attached to each camera for identification purpose.

QSAR modeling of different minimum potency levels for in vitro human CAR activation and inhibition and screening of 80,086 REACH and 54,971 U.S. substances

The human Constitutive Androstane Receptor (hCAR) is together with the human Pregnane X Receptor (hPXR) a key regulator of the metabolism and excretion of xenobiotics and endogenous compounds. Inhibition or activation of hCAR by xenobiotics can alter protein expression, leading to decreased or enhanced turnover of both xenobiotics and endogenous substances. Impacts from these alternations can potentially disturb physiological homeostasis and cause adverse effects. Tens-of-thousands of manufactured substances of which humans are potentially exposed are not tested for their potential to inhibit or activate hCAR. In this study, the U.S. Toxicology in the 21st Century (Tox21) high-throughput in vitro assay results for hCAR inhibition and activation were used in a comprehensive in-house process to derive training sets for different potency cut-offs, and to develop suites of quantitative structure–activity relationship (QSAR) models with binary outputs. Final, expanded models, which include substances from the external validation sets, were developed for select minimum potency levels. Rigorous cross- and external validations demonstrated good predictive accuracies for the models. The final models were applied to screen 80,086 EU and 54,971 U.S. substances, and the models predicted around 60% of the substances within their respective applicability domains (AD). Finally, statistical comparisons of hCAR predictions and QSAR predictions for a number of other endpoints related to e.g. Pregnane X, aryl hydrocarbon, estrogen and androgen receptors, as well as genotoxicity, cancer, sensitization and teratogenicity from the Danish (Q)SAR database were made to explore possible implications related to hCAR antagonists and agonists. The final models from this study will be made available in the free Danish (Q)SAR Models website. Predictions made with models from this study for 650,000 substances will be made available in the free Danish (Q)SAR Database. Predictions from the models developed in this study can for example contribute to priority setting, read-across cases and weight-of-evidence assessments of chemicals.

Unraveling the Signaling Balance of Activation and Exhaustion of CAR T Cells

CAR Tcells with different costimulation domains have proven clinical efficacy in leukemia and lymphoma but have different kinetics of activation, antigen sensitivity, and susceptibility to exhaustion. Two recent studies identified that these functions are shaped by a balance among opposing signaling complexes and transcription factors competing for binding motifs.

Changes in travel mode use over the life course with partner interactions in couple households

Abstract The paper links the mobility biography approach with studies on intra-household interactions between partners in travel. It does so by developing a multi-group path model – a structural equation model without latent variables – of changes in car use over time as a function of changes in activity and trip patterns, path dependency (baseline car use), life course changes, and the respective respondent's partner's car use, activity and trip patterns. In terms of life course changes, the paper accounts for (gradual, rather than abrupt) changes in paid and unpaid worktime, and the birth of a first or higher-order (i.e. second or further) child. The model further categorises respondents by gender. It focuses on couples who share one car. A model for multi-car households is estimated for comparison. The results show multiple interactions between two partners at the baseline as well as over time. The effects of the birth of a child are less clear than expected. Generally, they suggest that increases in driving are primarily an outcome of higher-order births, especially in one-car households. Policy conclusions are (1) that it may be overly optimistic to assume that the birth of a child can be utilised as a 'window of opportunity' to reduce driving, and (2) that social interactions between partners should be taken into account in policy concepts.

АКТУАЛЬНЫЕ ПРОБЛЕМЫ РАЗВИТИЯ АВТОСЕРВИСА В АЗЕРБАЙДЖАНЕ В СОВРЕМЕННЫХ УСЛОВИЯХ

The article deals with the actual problems of the development of car service in Azerbaijan in modern conditions. The important elements of ensuring the mobility of the personnel of car service enterprises and their effectiveness are analyzed. Ways of solving urgent problems of the service sector and organizing the activities of a car service in the context of the growth of consumer demand are considered. The factors of ensuring the mobility of the personnel of enterprises and the issues of their efficiency, as well as the availability of car service are taken into account. The necessity of significant improvement of the existing mechanisms and principles of management is substantiated, as well as the importance of decision-making towards the effective organization of car service. The importance of modeling and improving the activities of car service enterprises in accordance with the procedures and rules for their adequate functioning is considered. The necessity of organizing electronic and remote services, consultations and procedures for fulfilling orders with a visit to the addresses of clients has been substantiated. The conceptual elements for improving car service in the context of maximum customer satisfaction are considered. Based on the requirements of modernity, the importance of changing the attitude towards the offered services towards enhancing their quality and availability is revealed. The efficiency and creativity of the offered car repair services and preventive measures as a preventive technical inspection, revision of important parts and leading parts of cars were especially noted. The features of the service sector are analyzed, including the features of the car repair sector and the problems of this area. The issues of formation and improvement of the professional level of personnel are considered. The problems of transparency of car service activities and strengthening of its material and technical base have been investigated. A number of activities in the context of a pandemic and in the post-pandemic period have been proposed. The preparation of a package of car service services based on new approaches has been substantiated. The forms and mechanisms of enhancing the efficiency and mobility of car service enterprises in Azerbaijan in the context of a pandemic have been analyzed. More promising directions of increasing the activity of the personnel of car service enterprises are indicated. The conditions for the formation of services based on high technologies and their provision to end users are considered. Recommendations were prepared and proposals were made on topical problems and the development of car service in Azerbaijan in modern conditions.

Quantitative evaluation on parking configuration impact factor for promotion of private traffic in Zhangzhou ancient city historic district

As an important part of cultural heritage protection system, the protection of historic district seems to contradict with the inevitable motorised development of modern cities. However, in order to adapt to modern use, optimise and improve the traffic system and parking configuration of historic district based on goal of preservation and development may after all be accepted as a kind of alternative approach. There are many evidences shows that an appropriate optimisation on traffic system and parking configuration can enhance the connectivity and pedestrian-friendly environment, improve district dynamism, make tourists more satisfied, increase commercial activities and rental return. In addition, through the case study and analysis of Zhangzhou ancient city historic district, the paper finds that the positive elements of parking configuration factors are also reflected in a promotion of car owners' commercial activities and diverse consume content. The result plays a positive role in further understanding the optimisation of parking configuration enhance dynamism of historic district.

Modelling and Performance Analysis of Electric Car-Sharing Systems Using Petri Nets

Car sharing systems emerged as a new answer to mobility challenges in smart and sustainable cities. Despite their apparent success, design and exploitation of such systems raise crucial strategic and operational challenges. To help planners and decision makers, simulation, analysis and optimization models are unavoidable. Based on the formal modelling and analysis power of stochastic Petri nets, this paper proposes a discrete event simulation model for electric car sharing systems for performance and analysis purposes, taking into account their complex dynamic behaviour, organization and parameters including capacities of the stations, battery and energy availability, locations of charging stations and also their car maintenance activities, not negligible compared to the case of bike-sharing systems.

Quantitative Evaluation on Parking Configuration Impact Factor for Promotion of Private Traffic in Zhangzhou Ancient City Historic District

As an important part of cultural heritage protection system, the protection of historic district seems to contradict with the inevitable motorised development of modern cities. However, in order to adapt to modern use, optimise and improve the traffic system and parking configuration of historic district based on goal of preservation and development may after all be accepted as a kind of alternative approach. There are many evidences shows that an appropriate optimisation on traffic system and parking configuration can enhance the connectivity and pedestrian-friendly environment, improve district dynamism, make tourists more satisfied, increase commercial activities and rental return. In addition, through the case study and analysis of Zhangzhou ancient city historic district, the paper finds that the positive elements of parking configuration factors are also reflected in a promotion of car owners' commercial activities and diverse consume content. The result plays a positive role in further understanding the optimisation of parking configuration enhance dynamism of historic district.

Abstract B020: CAR-CD19 T cells secreting multi-antigen bridging proteins eliminate solid tumors

Abstract Introduction: Clinical data from diverse cell therapy trials highlight two critical issues. The first is resistance to, and relapse from, therapy due to antigen loss. In solid tumors antigen heterogeneity is common and cell therapies targeting a single antigen routinely fail as target antigen-negative tumor cells escape detection and emerge to drive tumor resistance. In hematologic malignancies, antigen loss after treatment drives relapses – up to 50% in B cell malignancies and near 100% in multiple myeloma. A related issue is that most cell therapeutics fail to persist after an initial expansion phase in the patient. This is particularly true in the case of solid tumors. This failure to persist may be due to unproductive engagement of the cell therapeutic with the target tumor cell. Here we describe the use of CAR-CD19 T cells (CAR19s) secreting bridging proteins to overcome these two issues. We leverage the intrinsic persistence properties of CAR19s and use these CARs to target multiple tumor antigens simultaneously using bridging proteins that link the CAR19s to multi-functional binding domains. Experimental Procedures: A stabilized form of the CD19 extracellular domain (ECD) (see Klesmith et al 2019) was cloned in frame with antigen binding domains, eg. scFv or VHH. We designed a core “bridging block” that consisted of an anti-Her2 scFv linked to the stabilized CD19 ECD to create a CD19-ECD-anti-Her2 bridging protein. This bridging protein sequence was then cloned into a lentiviral vector downstream of a CAR19 domain and a P2A cleavage site. Donor primary T cells transduced with the lentiviral particles expressed the CAR19 and secreted the bridging protein. The activities of the stabilized CD19-anti-Her2 bridging protein and the CAR19 T cells secreting that bridging protein were assessed using standard cytotoxicity assays in vitro and in vivo. Finally, we added an anti-EGFR scFv, thus creating a multi-antigen targeting bridging protein. We also created a version having a protease-sensitive mask to reduce EGFR binding outside of the acidic tumor microenvironment. Results: Use of the stabilized CD19 ECD in bridging proteins improves protein expression. Here we show for the first time that this improvement extends to bridging protein expression from CAR19 T cells. CAR19 T cells that secrete the stabilized CD19-anti-Her2 bridging protein are highly potent in vitro and in vivo. The CD19-ECD-anti-Her2 bridging block was further modified with an anti-EGFR scFv. This novel multi-antigen targeting bridging protein can bind either or both targeted antigens and supports highly potent cytotoxicity against dual positive tumor cells thus addressing the issue of solid tumor antigen heterogeneity. Ongoing programs add a third domain in an indication-specific manner, eg. an anti-IL-13Ra2 binding domain. In all cases the intrinsic anti-CD19 CAR activity is preserved, providing these unique CARs with a tumor-independent and self-renewing antigen depot in CD19-positive normal B cells. Conclusions: Here we have created a robust system for targeting multiple tumor antigens simultaneously. Further the use of CAR19s supports CAR-T cell persistence independently of tumor antigen expression. This unique technology addresses critical issues in cell therapy using a potent technology whose modular nature allows for rapid program and pipeline development. Citation Format: Paul Rennert, Christine Ambrose, Lihe Su, Fay Dufort, Alyssa Birt, Tom Sanford, Lan Wu, Roy Lobb. CAR-CD19 T cells secreting multi-antigen bridging proteins eliminate solid tumors [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference on Molecular Targets and Cancer Therapeutics; 2019 Oct 26-30; Boston, MA. Philadelphia (PA): AACR; Mol Cancer Ther 2019;18(12 Suppl):Abstract nr B020. doi:10.1158/1535-7163.TARG-19-B020

CRISPR-Cas9 disruption of PD-1 enhances activity of universal EGFRvIII CAR T cells in a preclinical model of human glioblastoma

Despite remarkable success in the treatment of hematological malignancies, CAR T-cell therapies for solid tumors have floundered, in large part due to local immune suppression and the effects of prolonged stimulation leading to T-cell dysfunction and exhaustion. One mechanism by which gliomas and other cancers can hamper CAR T cells is through surface expression of inhibitory ligands such as programmed cell death ligand 1 (PD-L1). Using the CRIPSR-Cas9 system, we created universal CAR T cells resistant to PD-1 inhibition through multiplexed gene disruption of endogenous T-cell receptor (TRAC), beta-2 microglobulin (B2M) and PD-1 (PDCD1). Triple gene-edited CAR T cells demonstrated enhanced activity in preclinical glioma models. Prolonged survival in mice bearing intracranial tumors was achieved after intracerebral, but not intravenous administration. CRISPR-Cas9 gene-editing not only provides a potential source of allogeneic, universal donor cells, but also enables simultaneous disruption of checkpoint signaling that otherwise impedes maximal antitumor functionality.

Blockade of CD95/CD95L Death Signaling Enhances CAR T Cell Persistence and Antitumor Efficacy

Introduction Despite the encouraging outcome of anti-CD19 chimeric antigen receptor T (CAR T) cell therapy in patients with B cell malignancies, CAR T cell persistence remains a major clinical challenge. Activation-induced cell death (AICD) is a programmed cell death caused by the interaction of CD95 and CD95L. Through specific blocking of the CD95-CD95L pathway, the CD95L inhibitor APG101 (Asunercept, Apogenix AG, Heidelberg) could prevent activated T cells from AICD. APG101 is a fully human fusion protein consisting of the extracellular domain of CD95 receptor and the Fc domain of an IgG antibody. Thus, we evaluated whether a blockade of the CD95L pathway through APG101 might improve CAR T cell persistence and enhance antitumor efficacy. Methods Human peripheral blood mononuclear cells (PBMCs) from healthy donors were stimulated by plate-bound CD3 and CD28 antibodies, and thereafter transduced with a 3rd generation CD19.CAR.CD28.CD137zeta retroviral vector. An in vitro co-culture stress test assay was employed to assess the functional status and viability of CD19.CAR T cells upon repetitive stimulation with CD19+ tumor cells, i.e. Daudi cells. CAR T cells (5.0 x 105 per well) were co-cultured with tumor cells at a 1:1 E:T ratio (round I) in the presence of APG101. Additional tumor cells were supplied to the co-culture every 24 hours. After 3 rounds (72 hr) of stimulation, tumor cells (CD3-CD19+) and CAR T cells (CD3+CD19-) were harvested for FACS analysis. To assess the antigen-induced CAR T cell proliferation, CAR T cells were preloaded with Cell Trace Violet cytosolic dye and cocultured with tumor cells for 72 hours. Results Activation-induced cell death of CAR T cells was observed after repeated antigenic stimulation, accompanied by increased CD95L expression. CD4+ CAR T cells were more susceptible to AICD compared to CD8+ CAR T cells. But, there was no difference in the expression of CD95L between CD4+ and CD8+ CAR T cells. Interestingly, addition of APG101 significantly inhibited CD95L expression and resulted in a lower level of CAR T cell death. Importantly, APG101 did not hamper the activation and proliferation of CAR T cells but was able to restore CAR T cell viability. The expression of PD1, TIM3 and LAG3 were also up-regulated after successive stimulation, however, their expression on CAR T cells were not influenced by APG101. After 3 days of co-culture, the number of CAR T cells increased in the presence of APG101 (7.9 x 105 vs6.0 x 105, P = 0.01) and residual tumor cells were diminished (1.7 x 105 vs2.7 x 105, P = 0.02). Of note, APG101 itself did not affect CAR T cells or tumor cells when cultured separately. Moreover, the central memory CAR T (TCM) cell subset showed higher CD95L expression after coculturing which could be inhibited by APG101. Therefore, the addition of APG101 to the coculture resulted in a significant accumulation of TCM subset after APG101 treatment. Conclusion Upregulation of CD95L after repeated antigen stimulation was reversed by APG101. CD95L blockade enhanced CAR T cell survival and promoted killing of tumor cells in vitro. Combining CAR T cell therapy with CD95L inhibitor might improve CAR T cell persistence in vivo and thus enhance the effect of CAR T cell therapy. Disclosures Schmitt: Therakos Mallinckrodt: Other: Financial Support . Kneisel:Apogenix AG: Employment. Hoeger:Apogenix AG: Employment, Membership on an entity's Board of Directors or advisory committees. Schmitt:MSD: Membership on an entity's Board of Directors or advisory committees, Other: Sponsoring of Symposia; Therakos Mallinckrodt: Other: Financial Support.

Mutation of the CD28 Co-Stimulatory Domain Confers Enhanced CAR T Cell Function

An obstacle with continued clinical development of CAR T cells is the limited understanding of their biology and mechanisms of anti-tumor immunity. We and others have shown that CARs with a CD28 co-stimulatory domain drive high levels of T cell activation that also lead to exhaustion and shortened persistence. The CD28 domain includes 3 intracellular subdomains (YMNM, PRRP, and PYAP) that regulate signaling pathways post TCR-stimulation, but it is unknown how they modulate activation and/or exhaustion of CAR T cells. A detailed understanding of the mechanism of CD28-dependent exhaustion in CAR T cells will allow the design of a CAR less prone to exhaustion and reduce relapse rates. This led us to hypothesize that by incorporating null mutations of CD28 subdomains (Fig 1A) we could optimize CAR T cell signaling and reduce exhaustion. In vitro, we found mutated CAR T cells with only a functional PYAP (mut06) subdomain secrete significantly less IFNγ, IL6, and TNFα after 24hr stimulation compared to non-mutated CD28 CAR T cells, but greater than the 1st generation m19z CAR. Also, cytotoxicity was enhanced compared to non-mutated CARs (Fig 1B). Using a pre-clinical immunocompetent mouse tumor model, we found the mut06 CAR T cell treated mice had a significant survival advantage compared to non-mutated CD28 CAR T cells (Fig 1C). To examine exhaustion, we ex vivo stimulated CAR T cells with target cells expressing CD19 and PDL1 and found mut06 CAR T cells had increased IFNγ (42%), TNFα (62%) and IL2 (73%) secretion compared to exhausted non-mutated CD28 CAR T cells. This suggests that mut06 CAR T cells are more resistant to exhaustion. To find a mechanistic explanation for this observation we examined CAR T cell signaling. After 24hr stimulation with CD19 target cells mut06 CAR T cells had a significant reduction in pAkt compared to m1928z CAR T cells, which is a critical signaling mediator in the NFAT and NR4A1 transcription factor pathways. Additionally, mut06 had decreased p-NFAT compared to m1928z when examined by western blot. To determine how optimized CAR signaling affected T cell exhaustion we looked at 22 genes that are upregulated when NFAT is constitutively active and overlap with genes identified as important for T cell exhaustion. We found that most of the exhaustion related genes were upregulated in m1928z CAR T cells while they were decreased in m19hBBz. The mut06 CAR T cell gene expression pattern was more similar to m19hBBz with exhaustion related genes downregulated compared to m1928z (Fig 1D). To examine differences in the accessibility of exhaustion related genes we performed ATAC-seq and found NFAT (Nfatc1) and NR4A2 (Nr4a2) had lower chromatin accessibility profiles in mut06 compared to m1928z (Fig 1E). We also found that exhaustion related genes Havcr2 (TIM3), Pdcd1 (PD1), and Lag3 (LAG3) all had greatly reduced chromatin accessibility in mut06 CAR T cells compared m1928z. Overall, these genomic studies support our findings that mut06 optimizes CAR T cell signaling by lowering transcription factors that regulate exhaustion. Figure 1 Disclosures Li: ImmuneBro Therapeutics: Other: sole shareholder . Davila:Atara: Research Funding; Celgene: Research Funding; GlaxoSmithKline: Consultancy; Novartis: Research Funding; Anixa: Consultancy; Bellicum: Consultancy; Adaptive: Consultancy; Precision Biosciences: Consultancy.

Chimeric Antigen Receptor T Cells Targeting NKG2D-Ligands Show Robust Efficacy Against Acute Myeloid Leukemia and T-Cell Acute Lymphoblastic Leukemia

CAR T cell approaches to effectively target AML and T-ALL without off-tumor effects on healthy myeloid or T cell compartments respectively are an unmet medical need. NKG2D-ligands are a promising target given their absence on healthy cells and surface expression in a wide range of malignancies. NKG2D-ligand expression in T-ALL has not previously been interrogated, and reports regarding the prevalence and density of NKG2D-ligand expression in AML vary. A first-in-human clinical trial of CART cells targeting NKG2D ligands in patients with AML demonstrated safety and feasibility (Baumeister et al, Cancer Immunology Research 2019). However, at low CAR T cell doses without lymphodepletion clinical efficacy was limited and in this selective patient cohort, NKG2D-ligand expression was only detectable at low levels. To broadly assess the level of NKG2D-ligand expression in AML and T-ALL and evaluate how this correlates with leukemia susceptibility to CAR T cells targeting NKG2D-ligands, we employed NKG2D-fusion protein-based flow-cytometry and RNA sequencing. CAR T cells bearing the full-length native NKG2D-receptor coupled to the CD3ζ cytoplasmic domain were generated using γ-retroviral transduction and then evaluated in functional assays. We show that NKG2D-ligands are expressed in T-ALL cell lines and primary T-ALL and confirm that NKG2D-ligands are frequently expressed at the surface and RNA-level in primary AML, albeit at relatively low levels. We demonstrate that high-expressing AML and T-ALL targets are exquisitely susceptible to in vitro activity of CAR T cells targeting NKG2D-ligands as evidenced by CD107a degranulation, cytotoxicity and production of IFN-γ and TNF-α. Importantly, we show that even low-level ligand expression in primary leukemia targets results in robust NKG2D-CAR activity. Using mass-cytometry, we reveal a distinct phenotypic profile of NKG2D-CAR T cells following incubation with ligand-positive leukemia targets when compared to controls. This includes selective upregulation of ICOS, 41BB, PD-1, LAG-3 and OX40. Given clinical precedent of pharmacologic HDAC inhibition in AML, we next assessed the impact of preincubation with Valproic Acid (VPA) on NKG2D-ligand expression in AML and whether this enhances susceptibility to NKG2D-CART cells in functional assays. We demonstrate that NKG2D-ligand expression can be selectively enhanced in low-expressing AML cell lines and primary AML blasts when AML cells are pre-incubated with 1mM VPA for 24 hours. Such pharmacologic ligand induction significantly augments NKG2D-CAR anti-leukemic activity in functional assays without affecting healthy Peripheral Blood Mononuclear Cells or CAR T cells. Our findings demonstrate that NKG2D-ligands are viable targets for CAR T cell therapy in AML and T-ALL despite low-level cell-surface expression and provide the rationale for the combination of HDAC-inhibitors with NKG2D-CAR T cell therapy as a potential strategy to achieve clinical NKG2D-CAR T cell efficacy in AML. Figure: A) Specific fluorescence intensity of NKG2D-ligand surface expression in Cell lines and Primary Leukemia cells are displayed in comparison B) NKG2D-CAR T cells (CAR) degranulate and produce proinflammatory cytokines during co-culture with AML and T-ALL lines (representative flow plot, left panel), but not to negative controls, whereas T cells transduced with vector lacking the CAR-transgene (Empty) do not. C) NKG2D-CAR T cells but not Empty control T cells display striking killing activity against AML and T-ALL cell lines. D) Preincubation of primary AML blasts with 1mM VPA for 24 hours enhances NKG2D-ligand expression and significantly augments NKG2D-CAR T cell activity, shown here as IFN-γ production during co-culture with 3 independent primary AML samples. Figure Disclosures Dranoff: Novartis: Employment. Ritz:Equillium: Research Funding; Merck: Research Funding; Kite Pharma: Research Funding; Aleta Biotherapeutics: Consultancy; Celgene: Consultancy; Avrobio: Consultancy; LifeVault Bio: Consultancy; Draper Labs: Consultancy; Talaris Therapeutics: Consultancy; TScan Therapeutics: Consultancy.

Bispecific Antibodies Enable Synthetic Agonistic Receptor-Transduced T Cells for Tumor Immunotherapy.

Genetically engineered T cells are powerful anticancer treatments but are limited by safety and specificity issues. We herein describe an MHC-unrestricted modular platform combining autologous T cells, transduced with a targetable synthetic agonistic receptor (SAR), with bispecific antibodies (BiAb) that specifically recruit and activate T cells for tumor killing. BiAbs of different formats were generated by recombinant expression. T cells were retrovirally transduced with SARs. T-cell activation, proliferation, differentiation, and T-cell-induced lysis were characterized in three murine and human tumor models in vitro and in vivo. Murine T cells transduced with SAR composed of an extracellular domain EGFRvIII fused to CD28 and CD3ζ signaling domains could be specifically recruited toward murine tumor cells expressing EpCAM by anti-EGFRvIII × anti-EpCAM BiAb. BiAb induced selective antigen-dependent activation, proliferation of SAR T cells, and redirected tumor cell lysis. Selectivity was dependent on the monovalency of the antibody for EGFRvIII. We identified FAS ligand as a major mediator of killing utilized by the T cells. Similarly, human SAR T cells could be specifically redirected toward mesothelin-expressing human pancreatic cancer cells. In vivo, treatment with SAR T cells and BiAb mediated antitumoral activity in three human pancreatic cancer cell xenograft models. Importantly, SAR activity, unlike CAR activity, was reversible in vitro and in vivo. We describe a novel ACT platform with antitumor activity in murine and human tumor models with a distinct mode of action that combines adoptive T-cell therapy with bispecific antibodies.

Modelling the impacts of EU countries\u2019 electric car deployment plans on atmospheric emissions and concentrations

The purpose of this work is to quantify key environmental impacts of electric vehicles deployment in the European Union. This is achieved by soft-linking three models (PRIMES-TREMOVE, DIONE and SHERPA) to explore a base and an alternative scenario. The alternative scenario draws on the assessment of the national policy frameworks for alternative fuels infrastructure requested by the Directive (2014/94/EU). Five environmental indicators are examined: tailpipe CO2, NOx and PM2.5 emissions as well as NO2 and PM2.5 urban background concentrations. By 2030, car travel activity is simulated to generate ca. 425 MtCO2/year in the EU28 under the alternative scenario. Compared to the base scenario, electric vehicles contribute to a 3% reduction in tailpipe CO2 emissions. Only two countries attain CO2 emission reductions greater than 10% in the model. The need for a higher level of policy ambition towards the deployment of less polluting vehicles in Europe is highlighted as a conclusion.

Modeling Future Energy Demand and CO2 Emissions of Passenger Cars in Indonesia at the Provincial Level

The high energy demand and CO2 emissions in the road transport sector in Indonesia are mainly caused by the use of passenger cars. This situation is predicted to continue due to the increase in car ownership. Scenarios are arranged to examine the potential reductions in energy demand and CO2 emissions in comparison with the business as usual (BAU) condition between 2016 and 2050 by controlling car intensity (fuel economy) and activity (vehicle-km). The intensity is controlled through the introduction of new car technologies, while the activity is controlled through the enactment of fuel taxes. This study aims to analyze the energy demand and CO2 emissions of passenger cars in Indonesia not only for a period in the past (2010–2015) but also based on projections through to 2050, by employing a provincially disaggregated bottom-up model. The provincially disaggregated model shows more accurate estimations for passenger car energy demands. The results suggest that energy demand and CO2 emissions in 2050 will be 50 million liter gasoline equivalent (LGE) and 110 million tons of CO2, respectively. The five provinces with the highest CO2 emissions in 2050 are projected to be West Java, Banten, East Java, Central Java, and South Sulawesi. The projected analysis for 2050 shows that new car technology and fuel tax scenarios can reduce energy demand from the BAU condition by 7.72% and 3.18% and CO2 emissions by 15.96% and 3.18%, respectively.

Putting CAR T cells in idle

Cancer Chimeric antigen receptor T (CAR T) cells can be an effective cell therapy for cancer. Unfortunately, excessive activation of CAR T cells can occasionally cause severe, even lethal, toxicity. Existing approaches can suppress overactive CAR T cells, but these generally kill the CAR T cells, which abrogates both their toxicity and their antitumor effects. In contrast, Mestermann et al. identified dasatinib as a drug that can temporarily inactivate CAR T cells. This helps reduce acute toxicity and allows the T cells to recover their anti-tumor effects after the drug is removed. Sci. Transl. Med. 11 , eaau5907 (2019).