LCL161 enhances expansion and survival of engineered anti-tumor T cells but is restricted by death signaling

BackgroundThe genesis of SMAC mimetic drugs is founded on the observation that many cancers amplify IAP proteins to facilitate their survival, and therefore removal of these pathways would re-sensitize the cells towards apoptosis. It has become increasingly clear that SMAC mimetics also interface with the immune system in a modulatory manner. Suppression of IAP function by SMAC mimetics activates the non-canonical NF-κB pathway which can augment T cell function, opening the possibility of using SMAC mimetics to enhance immunotherapeutics.MethodsWe have investigated the SMAC mimetic LCL161, which promotes degradation of cIAP-1 and cIAP-2, as an agent for delivering transient costimulation to engineered BMCA-specific human TAC T cells. In doing so we also sought to understand the cellular and molecular effects of LCL161 on T cell biology.ResultsLCL161 activated the non-canonical NF-κB pathway and enhanced antigen-driven TAC T cell proliferation and survival. Transcriptional profiling from TAC T cells treated with LCL161 revealed differential expression of costimulatory and apoptosis-related proteins, namely CD30 and FAIM3. We hypothesized that regulation of these genes by LCL161 may influence the drug’s effects on T cells. We reversed the differential expression through genetic engineering and observed impaired costimulation by LCL161, particularly when CD30 was deleted. While LCL161 can provide a costimulatory signal to TAC T cells following exposure to isolated antigen, we did not observe a similar pattern when TAC T cells were stimulated with myeloma cells expressing the target antigen. We questioned whether FasL expression by myeloma cells may antagonize the costimulatory effects of LCL161. Fas-KO TAC T cells displayed superior expansion following antigen stimulation in the presence of LCL161, suggesting a role for Fas-related T cell death in limiting the magnitude of the T cell response to antigen in the presence of LCL161.ConclusionsOur results demonstrate that LCL161 provides costimulation to TAC T cells exposed to antigen alone, however LCL161 did not enhance TAC T cell anti-tumor function when challenged with myeloma cells and may be limited due to sensitization of T cells towards Fas-mediated apoptosis

Tracking virus outbreaks in the twenty-first century

Emerging viruses have the potential to impose substantial mortality, morbidity and economic burdens on human populations. Tracking the spread of infectious diseases to assist in their control has traditionally relied on the analysis of case data gathered as the outbreak proceeds. Here, we describe how many of the key questions in infectious disease epidemiology, from the initial detection and characterization of outbreak viruses, to transmission chain tracking and outbreak mapping, can now be much more accurately addressed using recent advances in virus sequencing and phylogenetics. We highlight the utility of this approach with the hypothetical outbreak of an unknown pathogen, 'Disease X', suggested by the World Health Organization to be a potential cause of a future major epidemic. We also outline the requirements and challenges, including the need for flexible platforms that generate sequence data in real-time, and for these data to be shared as widely and openly as possible

Nitrogen and phosphorus addition to soil improves seed yield, foliar stomatal conductance, and the photosynthetic response of rapeseed (Brassica napus l.)

The effects of nitrogen and phosphorus levels on the physiological traits, yield, and seed yield of rapeseed (Brassica napus L.), were studied in a farm research project of Zanjan University. Three levels of nitrogen (0, 100, and 200 kg/ha) and three levels of phosphorus (0, 75, and 150 kg/ha) were considered. The results showed that an increase in nitrogen level caused an increase in the leaf chlorophyll content so that the application of 200 kg/ha of nitrogen increased the chlorophyll content of the leaves until the mid-grain filling stage. Nitrogen application lowered leaf stomatal conductance in the early flowering stage whereas the stomatal conductance was increased during the late flowering stage. Nitrogen application (100 and 200 kg/ha) also increased the quantum yield of photosystem II. On the other hand, with the application of 150 kg/ha and 75 kg/ha of phosphorus, the leaf stomatal conductance and the quantum yield of photosystem II in the early flowering stage increased respectively. The results showed that the application of 200 kg/ha of nitrogen and 75 kg/ha of phosphorus significantly increased seed and oil yield compared to the control. In addition, the number of siliques per plant and the weight of 1000 seeds showed an increasing trend that was affected by nitrogen and phosphorus levels. This study demonstrated that nitrogen enhanced the chlorophyll content, leaf area, and consequently, the quantum yield of photosystem II. Nitrogen also augmented the seed filling duration, seed yield, and oil yield by increasing gas exchange. As a result, the application of 100 kg/ha of nitrogen together with 75 kg/ha phosphorus showed the greatest effect on the qualitative and quantitative yield of rapeseed. However, the application of 200 kg/ha of nitrogen alone or in combination with different levels of phosphorus did not significantly increase many of the studied traits

Accelerated Quantitative 3D UTE-Cones Imaging Using Compressed Sensing

In this study, the feasibility of accelerated quantitative Ultrashort Echo Time Cones (qUTE-Cones) imaging with compressed sensing (CS) reconstruction is investigated. qUTE-Cones sequences for variable flip angle-based UTE T1 mapping, UTE adiabatic T1ρ mapping, and UTE quantitative magnetization transfer modeling of macromolecular fraction (MMF) were implemented on a clinical 3T MR system. Twenty healthy volunteers were recruited and underwent whole-knee MRI using qUTE-Cones sequences. The k-space data were retrospectively undersampled with different undersampling rates. The undersampled qUTE-Cones data were reconstructed using both zero-filling and CS reconstruction. Using CS-reconstructed UTE images, various parameters were estimated in 10 different regions of interests (ROIs) in tendons, ligaments, menisci, and cartilage. Structural similarity, percentage error, and Pearson’s correlation were calculated to assess the performance. Dramatically reduced streaking artifacts and improved SSIM were observed in UTE images from CS reconstruction. A mean SSIM of ~0.90 was achieved for all CS-reconstructed images. Percentage errors between fully sampled and undersampled CS-reconstructed images were below 5% for up to 50% undersampling (i.e., 2× acceleration). High linear correlation was observed (>0.95) for all qUTE parameters estimated in all subjects. CS-based reconstruction combined with efficient Cones trajectory is expected to achieve a clinically feasible scan time for qUTE imaging

Correlations between elastic modulus and ultrashort echo time (UTE) adiabatic T1ρ relaxation time (UTE-Adiab-T1ρ) in Achilles tendons and entheses

Patients with psoriatic arthritis commonly have abnormalities of their entheses, which are the connections between tendons and bone. There are shortcomings with the use of conventional magnetic resonance imaging (MRI) sequences for the evaluation of entheses and tendons, whereas ultrashort echo time (UTE) sequences are superior for the detection of high signals, and can also be used for non-invasive quantitative assessments of these structures. The combination of UTE-MRI with an adiabatic-T1ρ preparation (UTE-Adiab-T1ρ) allows for reliable assessment of entheses and tendons with decreased susceptibility to detrimental magic angle effects. This study aimed to investigate the relationship between quantitative UTE-MRI measures and the biomechanical properties of Achilles tendons and entheses. In total, 28 tendon-enthesis sections were harvested from 11 fresh-frozen human cadaveric foot-ankle specimens (52 ± years old). Tendon-enthesis sections were scanned using the UTE-Adiab-T1ρ and UTE-T1 sequences on a clinical 3 T scanner. MRI-based measures and indentation tests were performed on the enthesis, transitional, and tensile tendon zones of the specimens. Hayes' elastic modulus showed significant inverse correlations (Spearman's) with UTE-Adiab-T1ρ in all zones (R= - 0.46, - 0.54, and - 0.61 in enthesis, transition, and tensile tendon zones, respectively). Oliver-Pharr's elastic modulus showed significant inverse correlations with UTE-Adiab-T1ρ in transition (R= - 0.52) and tensile tendon zone (R=- 0.60). UTE-T1 did not show significant correlations with the elastic modulus. UTE-MRI and elastic modulus were significantly lower in the tensile tendon compared with the enthesis regions This study highlights the potential of the UTE-Adiab-T1ρ technique for the non-invasive evaluation of tendons and enthuses