Engineering homing properties of cancer-specific T lymphocytes in adoption cell therapy

Several cancer immunotherapies harness the ability of CD8 T cells to eradicate tumours. Adoptive cell transfer of cancer-specific T cells is an immunotherapy that has produced promising results for cancer patients. L-selectin is a membrane protein that enables T cell rolling on high endothelial cells and transendothelial migration into lymph nodes. During this process, and after T cell activation, the L-selectin ectodomain is proteolytically cleaved from the T cell surface by ADAM17. In preclinical melanoma models, adoptively transferred cancer-specific T cells, which expressed ectodomain shedding-resistant L-selectin, better controlled tumour growth than T cells bearing wildtype L-selectin. However, these adoptively transferred T cells were naïve and transgenic for a cancer-specific TCR and variants of L-selectin. This is a situation not reflected in the clinic. Here, I delivered a cancerspecific TCR, and variants of L-selectin, to T cells using a clinically relevant method. I further demonstrated that modified L-selectin variants affected CD8 T cell expression of CD25, Ki67, and PD-1 during cytolysis assays. A lack of L-selectin ectodomain shedding has also been implicated in degranulation and killing by effector T cells, as well as naïve T cell activation-mediated upregulation of CD25 and consequent T cell proliferation. Recently, we demonstrated that ADAM17-dependent L-selectin ectodomain proteolysis generates a membraneretained fragment (MRF) that undergoes γ-secretase proteolysis. The intracellular domain, or tail, of L-selectin interacts with several proteins, including PKC, grb-SOS, and lck. Therefore, the L-selectin tail may perform signalling functions prior to, and following, γ-secretase proteolysis in T cells. I demonstrated that the MRF of Lselectin is lost from the cell membrane, either through degradation by the proteasome or further proteolysis by γ-secretase. Therefore, the altered phenotypes conferred by shedding-resistant L-selectin to cancer-specific T cells are likely due to retention of the tail of L-selectin at the cell surface and continued interaction with binding partners

Meteorological characteristics and assessment of the effect of local emissions during high PM10 concentration in the Seoul Metropolitan Area

In this study, we investigate the meteorological characteristics and the effect of local emissions during high PM10 concentrations in the Seoul Metropolitan Area (SMA) by utilizing data from a high-resolution urban meteorological observation system network (UMS-Seoul) and The Air Pollution Model (TAPM). For a detailed analysis, days with PM10 concentrations higher than 80 ??g m-3 for daily average PM10 concentration (classified as unhealthy by the Korean Ministry of Environment) in the Seoul Metropolitan Area (SMA) were classified into 3 Cases. Case I was defined as when the prevailing effect was from outside the SMA. Case II was defined as when the prevailing effect was a local effect with outside. Case III was defined as when the prevailing effect was local. Overall, high PM10 concentrations in the SMA mostly occurred under weak migratory anticyclone systems over the Korean Peninsula during warm temperatures. Prior to the PM10 concentration reaching the peak concentration, the pattern in each case was distinctive. After peak concentrations, however, the pattern for the 3 cases became less distinct. This study showed that nearly 50% of the high PM10 concentrations in the SMA occurred in spring and were governed by the conditions for Case II more than these for Cases I and III. In spring, the main sources of the high PM10 concentrations in the SMA were local emissions due to the predominance of weak winds and local circulation. The simulation showed that the non-SMA emissions were about 63 to 73% contribution to the spring high PM10 concentrations in the SMA. Specifically, local point sources including industrial combustion, electric utility, incineration and cement production facilities scattered around the SMA and could account for PM10 concentrations more than 10 ??g m-3 in the SMA

Pharamcological inhibition of protein tyrosine phosphatase 1B protects against atherosclerotic plaque formation in LDLR-/- mouse model of atherosclerosis

The authors wish to thank Professor Nicholas Tonks for providing the PTP1B inhibitor trodusquemine; Linda Robertson for her help with the aorta histology; Dr Fiona Grieg for tuition into aortic dissection and Dr James Hislop for critical reading of this manuscript. We also wish to thank the British Heart Foundation (PG/14/43/30889) for supporting this researchPeer reviewedPublisher PD

Modelling the prestress transfer in pre-tensioned concrete elements

Three models were developed to simulate the transfer of prestress force from steel to concrete in pre-tensioned concrete elements. The first is an analytical model based on the thick-walled cylinder theory and considers linear material properties for both steel and concrete. The second is an axi-symmetric finite element (FE) model with linear material properties; it is used to verify the analytical model. The third model is a three dimensional nonlinear FE model. This model considers the post-cracking behaviour of concrete as well as concrete shrinkage and the time of prestress releasing. A new expression from the analytical model is developed to estimate the transmission length as well as the stress distribution along the tendon. The paper also presents a parametric study to illustrate the impact of diameter of prestressing steel, concrete cover, concrete strength, initial prestress, section size, surface roughness of prestressing steel, time of prestress release, and the member length on the transfer of stress in pre-tensioned concrete elements

ADAM17-dependent proteolysis of L-selectin promotes early clonal expansion of cytotoxic T cells

L-selectin on T-cells is best known as an adhesion molecule that supports recruitment of blood-borne naïve and central memory cells into lymph nodes. Proteolytic shedding of the ectodomain is thought to redirect activated T-cells from lymph nodes to sites of infection. However, we have shown that activated T-cells re-express L-selectin before lymph node egress and use L-selectin to locate to virus-infected tissues. Therefore, we considered other roles for L-selectin proteolysis during T cell activation. In this study, we used T cells expressing cleavable or non-cleavable L-selectin and determined the impact of L-selectin proteolysis on T cell activation in virus-infected mice. We confirm an essential and non-redundant role for ADAM17 in TCR-induced proteolysis of L-selectin in mouse and human T cells and show that L-selectin cleavage does not regulate T cell activation measured by CD69 or TCR internalisation. Following virus infection of mice, L-selectin proteolysis promoted early clonal expansion of cytotoxic T cells resulting in an 8-fold increase over T cells unable to cleave L-selectin. T cells unable to cleave L-selectin showed delayed proliferation in vitro which correlated with lower CD25 expression. Based on these results, we propose that ADAM17-dependent proteolysis of L-selectin should be considered a regulator of T-cell activation at sites of immune activity

Design of Experiments for Screening

The aim of this paper is to review methods of designing screening experiments, ranging from designs originally developed for physical experiments to those especially tailored to experiments on numerical models. The strengths and weaknesses of the various designs for screening variables in numerical models are discussed. First, classes of factorial designs for experiments to estimate main effects and interactions through a linear statistical model are described, specifically regular and nonregular fractional factorial designs, supersaturated designs and systematic fractional replicate designs. Generic issues of aliasing, bias and cancellation of factorial effects are discussed. Second, group screening experiments are considered including factorial group screening and sequential bifurcation. Third, random sampling plans are discussed including Latin hypercube sampling and sampling plans to estimate elementary effects. Fourth, a variety of modelling methods commonly employed with screening designs are briefly described. Finally, a novel study demonstrates six screening methods on two frequently-used exemplars, and their performances are compared

Understanding anti-government sentiment and the public's anti-corruption behavior: The roles of perceived ethics failure, emotional leadership, and communication strategy

This study conceptualizes anti-government sentiment and tests the relationships between anti-government sentiment and three antecedents (i.e., ethics failure, compassionate leadership, and communication strategy). An online survey (n=1,112) was conducted in South Korea. Exploratory Factor Analysis with Principal Component Analysis and Confirmatory Factor Analysis were conducted on the measures proposed for anti-government sentiment. The hypotheses were tested using Structural Equation Modeling. Results show that publics’ perceptions of the government’s ethics failure, lack of compassionate leadership, and use of a buffering strategy for communication are positively related to their anti-government sentiment toward the incumbent government

Impact Comparison of Synoptic Meteorology and Nationwide/local Emissions on the Seoul Metropolitan Area during High PM Multi-event and Non-event Days

Meteorology and emissions play very important roles in the concentrations of air pollutants during severe haze/smog periods. This study compares the impacts of synoptic meteorology and nationwide/local emissions during high PM10 multi-event and non-event days in the Seoul Metropolitan Area (SMA). The multi-event and non-event cases were selected based on daily mean PM10 concentrations in Seoul from January 2014 to March 2019. The multi-event cases in spring and winter were closely associated with weak synoptic winds, while that in autumn was due to the strong winds at the rear side of a strong cold front, which induced the Asian dust event in northeastern China and Korea. The multi-event case in spring was found to be mainly due to series of migratory anticyclones, while winter case was due to the stagnant system after northerly winds. The surrounding low pressure systems as well as high pressure systems could be important to determine whether the synoptic systems would be stagnant or not. The fractional contributions of SMA emissions to the mean PM10 and PM2.5 concentrations were 24%-35% and 22%-35% for the multi-event cases, respectively. The contributions to the maximum PM10 and PM2.5 concentrations were larger than those to the mean concentrations by 16%-23% and 19%-26% for the multi-event cases, respectively

Cloning and functional characterisation of polyunsaturated fatty acid elongases of marine and freshwater teleost fish

Enzymes that lengthen the carbon chain of polyunsaturated fatty acids are key to the biosynthesis of the highly unsaturated fatty acids, arachidonic, eicosapentaenoic and docosahexaenoic acids from linoleic and α-linolenic acids. A Mortierella alpina cDNA polyunsaturated fatty acid elongase sequence identified mammalian, amphibian, zebrafish and insect expressed sequence tags (ESTs) in GenBank. Consensus primers were designed in conserved motifs and used to isolate full length cDNA from livers of several fish species by Rapid Amplification of cDNA Ends (RACE). The amplified cDNAs encoded putative open reading frames (ORFs) of 288-294 amino acids that were highly conserved among the fish species. Heterologous expression in yeast, Saccharomyces cerevisiae, demonstrated that all of the ORFs encoded elongases with the ability to lengthen polyunsaturated fatty acid substrates with chain lengths from C18 to C22 and also monounsaturated fatty acids, but not saturated fatty acids. There were differences in the functional competence of the elongases from different fish species. Most of the fish elongases showed a pattern of activity towards different fatty acid substrates in the rank order C18 > C20 >C22, although the tilapia and turbot elongases had similar activity towards 18:4n-3 and 20:5n-3. The fish elongases generally showed greater activity or similar activities with n-3 than with n-6 homologues, with the exception of the cod enzyme which was more active towards n-6 fatty acids