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Cancer immunotherapies have achieved much success and have become the forefront treatment of cancers previously associated with poor prognosis. However, a major challenge in cancer immunotherapies ...

Background Immune checkpoint inhibitors (ICIs) have revolutionized the cancer therapeutic landscape and our perception of interactions between the immune system and tumor cells. Despite remarkable progress, disease relapse and primary resistance are not uncommon. Understanding the biological processes that tumor-infiltrating lymphocytes (TILs) undergo during ICI, how this affects the tumor microenvironment (TME) and, ultimately, clinical outcome is, therefore, necessary to further improve treatment efficacy. Aim In the current study, we sought to characterize TILs from patients with metastatic solid tumors undergoing ICI correlating flowcytometric findings with clinical outcome. Methods In total, 20 patients with 10 different metastatic solid tumors treated with ICIs targeting programmed-cell death-1 (PD-1)/PD-L1 axis were included in this study. The phenotype of T cells deriving from biopsies obtained prior to treatment initiation and on-treatment was investigaded. Analyses were focused on T cells’ degree of differentiation and activity and how they correlate with transcriptomic changes in the TME. Results Data indicate that patients benefitting from ICIs accumulate CD8+central memory T cells. TILs developed an effector-like phenotype over time, which was also associated with a cytolytic gene signature. In terms of modulation of T-cell responses, we observed that high expression of checkpoint molecules pre-treatment (i.e., PD-1, lymphocyte activation gene-3 [LAG-3], B and T-lymphocyte attenuator [BTLA] and T-cell immunoglobulin and mucin domain containing-3 [TIM-3]) was associated with similar gene signature and correlated to treatment benefit. Increasing expression of LAG-3 and BTLA in the CD8 compartment and their co-expression with PD-1 during treatment were, however, a common feature for patients who failed to respond to ICIs. Conclusions Besides identifying immune profiles suggestive of response to ICI, our results provide a more nuanced picture regarding expression of checkpoint molecules that goes beyond T-cell anergy.

This article qualitatively shows the yaw stability of a free-yawing downwind turbine and the ability of the turbine to align passively with the wind direction using a model with 2 degrees of freedom. An existing model of a Suzlon S111 upwind 2.1 MW turbine is converted into a downwind configuration with a 5∘ tilt and a 3.5∘ downwind cone angle. The analysis shows that the static tilt angle causes a wind-speed-dependent yaw misalignment of up to −19∘ due to the projection of the torque onto the yaw bearing and the skewed aerodynamic forces caused by wind speed projection. With increased cone angles, the yaw stiffness can be increased for better yaw alignment and the stabilization of the free-yaw motion. The shaft length influences the yaw alignment only for high wind speeds and cannot significantly contribute to the damping of the free-yaw mode within the investigated range. Asymmetric flapwise blade flexibility is seen to significantly decrease the damping of the free-yaw mode, leading to instability at wind speeds higher than 19 m s−1. It is shown that this additional degree of freedom is needed to predict the qualitative yaw behaviour of a free-yawing downwind wind turbine.

Despite encouraging results with immune checkpoint inhibition (ICI), a large fraction of cancer patients still does not achieve clinical benefit. Finding predictive markers in the complexity of the tumor microenvironment is a challenging task and often requires invasive procedures. In our study, we looked for putative variables related to treatment benefit among immune cells in peripheral blood across different tumor types treated with ICIs. For that, we included 33 patients with different solid tumors referred to our clinical unit for ICI. Peripheral blood mononuclear cells were isolated at baseline, 6 and 20 weeks after treatment start. Characterization of immune cells was carried out by multi-color flow cytometry. Response to treatment was assessed radiologically by RECIST 1.1. Clinical outcome correlated with a shift towards an effector-like T cell phenotype, PD-1 expression by CD8+T cells, low levels of myeloid-derived suppressor cells and classical monocytes. Dendritic cells seemed also to play a role in terms of survival. From these findings, we hypothesized that patients responding to ICI had already at baseline an immune profile, here called ‘favorable immune periphery’, providing a higher chance of benefitting from ICI. We elaborated an index comprising cell types mentioned above. This signature correlated positively with the likelihood of benefiting from the treatment and ultimately with longer survival. Our study illustrates that patients responding to ICI seem to have a pre-existing immune profile in peripheral blood that favors good outcome. Exploring this signature can help to identify patients likely to achieve benefit from ICI.

Frontiers in oncology 11, 637420 (2021). doi:10.3389/fonc.2021.637420 special issue: "Novel Treatment Strategies for Myeloproliferative Neoplasms / Edited by Ken-Hong Lim, Shinobu Matsuura, Bing Xu", Published by Frontiers Media, Lausanne

In this chapter, we cover the derivation of aero-servo-elastic models of wind turbines, the modal dynamics of two- and three -bladed turbines, and the aeroelastic stability of their operational states. Knowledge of the modal properties and the aeroelastic stability limits are fundamental requirements for a good turbine design without resonance and self-induced vibrations.

Coupling between bending and twist has a significant influence on the aeroelastic response of wind turbine blades. The coupling can arise from the blade geometry (e.g. sweep, prebending, or deflection under load) or from the anisotropic properties of the blade material. Bend–twist coupling can be utilized to reduce the fatigue loads of wind turbine blades. In this study the effects of material-based coupling on the aeroelastic modal properties and stability limits of the DTU 10 MW Reference Wind Turbine are investigated. The modal properties are determined by means of eigenvalue analysis around a steady-state equilibrium using the aero-servo-elastic tool HAWCStab2 which has been extended by a beam element that allows for fully coupled cross-sectional properties. Bend–twist coupling is introduced in the cross-sectional stiffness matrix by means of coupling coefficients that introduce twist for flapwise (flap–twist coupling) or edgewise (edge–twist coupling) bending. Edge–twist coupling can increase or decrease the damping of the edgewise mode relative to the reference blade, depending on the operational condition of the turbine. Edge–twist to feather coupling for edgewise deflection towards the leading edge reduces the inflow speed at which the blade becomes unstable. Flap–twist to feather coupling for flapwise deflections towards the suction side increase the frequency and reduce damping of the flapwise mode. Flap–twist to stall reduces frequency and increases damping. The reduction of blade root flapwise and tower bottom fore–aft moments due to variations in mean wind speed of a flap–twist to feather blade are confirmed by frequency response functions.

Background aims We investigated whether the addition of an autologous dendritic cell–based cancer vaccine (DCvac) induces an immune response in patients with metastatic castration-resistant prostate cancer treated with docetaxel. Methods Forty-three patients were randomized 1:1 to receive up to 10 cycles of docetaxel alone, 75 mg/m 2 every 3 weeks or in combination with DCvac. Monocytes were harvested following a leukapheresis procedure, matured ex vivo and subsequently transfected with messenger RNA encoding multiple tumor-associated antigens (TAAs). DCvac was administered intradermally twice through treatment cycles 1–4 and once through treatment cycles 5–10. Immune cell composition and antigen-specific responses were analyzed using flow cytometry, ELISpot and delayed type hypersensitivity (DTH) tests. Toxicity was graded according to Common Terminology Criteria for Adverse Events version 3.0. Progression-free survival (PFS) and disease-specific survival (DSS) was calculated using the Kaplan-Meier method. Results Prostate-specific antigen responses were similar in patients treated with docetaxel alone and combination therapy (58% versus 38%; P = 0.21). PFS and DSS were comparable: 5.5 versus 5.7 months ( P = 0.62, log rank) and 21.9 versus 25.1 months ( P = 0.60, log rank). Nine (50%) and 14 (78%) patients treated with docetaxel and DCvac had a TAA-specific or vaccine-specific immune response in the ELISpot and DTH analysis, respectively. Vaccine induced toxicity was limited to local reactions. Decline in myeloid-derived suppressor cells at the third treatment cycle was found to be an independent predictor of DSS. Conclusions The addition of DCvac was safe. Immune responses were detected in approximately half of the patients investigated.

This paper presents the integration of a near-wake model for trailing vorticity, which is based on a prescribed-wake lifting-line model proposed by Beddoes(1987), with a blade element momentum (BEM)-based far-wake model and a 2-D shed vorticity model. The resulting coupled aerodynamics model is validated against lifting-surface computations performed using a free-wake panel code. The focus of the description of the aerodynamics model is on the numerical stability, the computation speed and the accuracy of unsteady simulations. To stabilize the near-wake model, it has to be iterated to convergence, using a relaxation factor that has to be updated during the computation. Further, the effect of simplifying the exponential function approximation of the near-wake model to increase the computation speed is investigated in this work. A modification of the dynamic inflow weighting factors of the far-wake model is presented that ensures good induction modeling at slow timescales. Finally, the unsteady airfoil aerodynamics model is extended to provide the unsteady bound circulation for the near-wake model and to improve the modeling of the unsteady behavior of cambered airfoils. The model comparison with results from a free-wake panel code and a BEM model is centered around the NREL 5 MW reference turbine. The response to pitch steps at different pitching speeds is compared. By means of prescribed vibration cases, the effect of the aerodynamic model on the predictions of the aerodynamic work is investigated. The validation shows that a BEM model can be improved by adding near-wake trailed vorticity computation. For all prescribed vibration cases with high aerodynamic damping, results similar to those obtained by the free-wake model can be achieved in a small fraction of computation time with the proposed model. In the cases with low aerodynamic damping, the addition of trailed vorticity modeling shifts the results closer to those obtained by using the free-wake code, but differences remain.

The effects of bend–twist coupling on the aeroelastic modal properties and stability limits of a two-dimensional blade section in attached flow are investigated. Bend–twist coupling is introduced in the stiffness matrix of the structural blade section model. The structural model is coupled with an unsteady aerodynamic model in a linearised state–space formulation. A numerical study is performed using structural and aerodynamic parameters representative for wind turbine blades. It is shown that damping of the edgewise mode is primarily influenced by the work of the lift which is close to antiphase, making the stability of the mode sensitive to changes in the stiffness matrix. The aerodynamic forces increase the stiffness of the flapwise mode for flap–twist coupling to feather for downwind deflections. The stiffness reduces and damping increases for flap–twist to stall. Edge–twist coupling is prone to an edgetwist flutter instability at much lower inflow speeds than the uncoupled blade section. Flap–twist coupling results in a moderate reduction of the flutter speed for twist to feather and divergence for twist to stall.

The modal dynamics of structures with bladed isotropic rotors is analyzed using Hill's method. First, analytical derivation of the periodic system matrix shows that isotropic rotors with more than two blades can be represented by an exact Fourier series with 3/rev (three per rotor revolution) as the highest order. For two-bladed rotors, the inverse mass matrix has an infinite Fourier series with harmonic components of decreasing norm; thus, the system matrix can be approximated by a truncated Fourier series of predictable accuracy. Second, a novel method for automatically identifying the principal solutions of Hill's eigenvalue problem is introduced. The corresponding periodic eigenvectors can be used to compute symmetric and antisymmetric components of the two-bladed rotor motion, as well as the additional forward and backward whirling components for rotors with more than two blades. To illustrate the use of these generic methods, a simple wind turbine model is set up with three degrees of freedom for each blade and seven degrees of freedom for the nacelle and drivetrain. First, the model parameters are tuned such that the low-order modal dynamics of a three-bladed 10 MW turbine from previous studies is recaptured. Second, one blade is removed, leading to larger and higher harmonic terms in the system matrix. These harmonic terms lead to modal couplings for the two-bladed turbine that do not exist for the three-bladed turbine. A single mode of a two-bladed turbine will also have several resonance frequencies in both the ground-fixed and rotating frames of reference, which complicates the interpretation of simulated or measured turbine responses.

Our objective was to investigate if the tumor microRNA (miRNA) expression profile was related to tumor growth rate. Growth-related miRNAs might be potential targets for future therapeutic intervention. Tumor tissue was sampled during surgery of patients with a sporadic vestibular schwannoma. Tumor growth rate was determined by tumor measurement on the two latest pre-operative MRI scans. Tumor miRNA expression was analyzed using the Affymetrix Gene Chip® protocol, and CEL files were generated using GeneChip® Command Console® Software and normalized using Partek Genomics Suite 6.5. The CEL files were analyzed using the statistical software program R. Principal component analysis, affected gene ontology analysis, and analysis of miRNA expression fold changes were used for analysis of potential relations between miRNA expression profile and tumor growth rate. Tumor miRNA expression is related to the growth rate of sporadic vestibular schwannomas. Rapid tumor growth is associated with deregulation of several miRNAs, including upregulation of miR-29abc, miR-19, miR-340-5p, miR-21, and miR-221 and downregulation of miR-744 and let-7b. Gene ontologies affected by the deregulated miRNAs included neuron development and differentiation, gene silencing, and negative regulation of various biological processes, including cellular and intracellular signaling and metabolism.

PD-L1-specific T cells are a natural part of the T-cell repertoire in humans. Hence, we have previously described spontaneous CD8+ and CD4+ T-cell reactivity against PD-L1 in the peripheral blood of patients with various cancers as well as in healthy donors. It is well described that the expression of the PD-L1 protein is introduced in cells by pro-inflammatory cytokines, e.g. IFN-γ. In the current study, we were able to directly link inflammation with PD-L1-specific T cells by showing that inflammatory mediators such as IFN-γ generate measurable numbers of PD-L1-specific T cells in human PBMCs as well as in in vivo models. These PD-L1-specific T cells can vigorously modulate the cell compartments of the local environment. PD-L1-specific T cells may be important for immune homeostasis by sustaining the ongoing inflammatory response by the suppression of regulatory cell function both directly and indirectly.

This work considers the design driving load cases from a full design load basis analysis on an upwind turbine changed into a downwind configuration. The upwind turbine is a commercial class IIIA 2.1-MW turbine, manufactured by Suzlon. The downwind turbine shows an increase in the normalized tower clearance by 6%, compared with the upwind concept. Removing the blade prebend increases the normalized minimum tower clearance by 17% in the downwind configuration compared with the upwind configuration. The extreme loads on the longitudinal tower bottom bending moment are seen to generally increase by 17% because of the overhanging gravity moment of the rotor-nacelle assembly. The extreme blade root bending moments are reduced by 10% flapwise, because of the coning of the rotor in downwind direction. The fatigue loads suffer from the tower shadow, leading to an overall increase of the fatigue loads in the blades with up to 5% in flapwise direction in the downwind configuration. Because of blade deflection and coning direction, the downwind configuration shows a 0.75% lower annual energy production. Removing the prebend increases the annual energy production loss to 1.66%.

Background Excessive extracellular matrix (ECM) remodeling and a reactive stroma can affect T-cell infiltration and T-cell activity in the tumor and hereby influence response to immune checkpoint inhibitors (ICI). In the pursuit of finding biomarkers that predict treatment response, we evaluated the association between serum biomarkers of collagen and vimentin turnover and outcomes in metastatic melanoma patients treated with the anti-CTLA-4 antibody ipilimumab (IPI). Methods Type III collagen formation (PRO-C3), MMP-degraded type I, type III and type IV collagens (C1M, C3M and C4M), and citrullinated and MMP-degraded vimentin (VICM) were measured with ELISAs in serum from metastatic melanoma patients before (n = 66) and 3 weeks after (n = 52) initiation of IPI treatment. Biomarker levels were associated with Disease Control Rate (DCR) and survival outcomes. Results We found that baseline levels of PRO-C3 (p = 0.011), C1M (p = 0.003), C3M (p = 0.013) and C4M (p = 0.027) were significantly elevated in patients with progressive disease (PD). Univariate Cox regression analysis identified high PRO-C3 (p = 0.021) and C4M (p = 0.008) as predictors of poor overall survival (OS) and the biomarkers remained significant when evaluated with other covariates (PRO-C3 (p = 0.049) and C4M (p = 0.046)). Multivariate analysis identified VICM as a predictor of longer OS (p = 0.026). Similarly, a high C3M/PRO-C3 ratio predicted for increased OS (p = 0.034). Only C3M (p = 0.003) and VICM (p

A multi-rotor wind turbine (MRWT) is accepted as a novel solution in reducing the rotor size without decreasing the energy harvesting capability compared to a single-rotor wind turbine (SRWT). However, there is no systematic methodology available for obtaining the modal properties of MRWTs. This paper presents a novel methodology and systematic modeling method of modal dynamics with comprehensive numerical simulations and analyses of MRWTs. In the methodology, Coleman transformation is employed to convert the nonlinear time-variant dynamics formulation of a MRWT into a set of linear time-invariant dynamic equations. Using HAWC2 and HAWCStab2 a method to establishing high-fidelity linear time-invariant dynamic models is presented. The structural modal dynamics of a tri-rotor wind turbine is systematically investigated by using modal analysis theory and interpreting the rotor speed-dependent dynamics visualized in a Campbell diagram. The investigation shows that rotor modes on a MRWT have similar behavior as for a SRWT and that low torsional stiffness of the rotor attachment point affects the first flapwise backward whirling modes by decreasing the corresponding natural frequencies. Moreover, the low torsional stiffness results in a delayed splitting of the asymmetric flapwise rotor modes. The simulations and analyses result verified the proposed dynamic modeling method. The major contributions in this paper provide the essential insights and guidance in the design of MRWTs but also for SRWT with slender towers.

Though present in low numbers, dendritic cells (DCs) are recognized as major players in the control of cancer by adaptive immunity. The roles of cytotoxic CD8+ T-cells and Th1 helper CD4+ T-cells are well-documented in murine models of cancer and associated with a profound prognostic impact when infiltrating human tumors, but less information is known about how these T-cells gain access to the tumor or how they are primed to become tumor-specific. Here, we highlight recent findings that demonstrate a vital role of CD103+ DCs, which have been shown to be experts in cross-priming and the induction of anti-tumor immunity. We also focus on two different mediators that impair the function of tumor-associated DCs: prostaglandin E2 and β-catenin. Both of these mediators seem to be important for the exclusion of T-cells in the tumor microenvironment and may represent key pathways to target in optimized treatment regimens against cancer.

A high-fidelity linear time-invariant model of the aero-servo-elastic response of a wind turbine with trailing-edge flaps is presented and used for systematic tuning of an individual flap controller. The model includes the quasi-steady aerodynamic effects of trailing-edge flaps on wind turbine blades and is integrated in the linear aeroelastic code HAWCStab2. The dynamic response predicted by the linear model is validated against non-linear simulations, and the quasi-steady assumption does not cause any significant response bias for flap deflection with frequencies up to 2–3 Hz. The linear aero-servo-elastic model support the design, systematic tuning and model synthesis of smart rotor control systems. As an example application, the gains of an individual flap controller are tuned using the Ziegler–Nichols method for the full-order poles. The flap controller is based on feedback of inverse Coleman transformed and low-pass filtered flapwise blade root moments to the cyclic flap angles through two proportional-integral controllers. The load alleviation potential of the active flap control, anticipated by the frequency response of the linear closed-loop model, is also confirmed by non-linear time simulations. The simulations report reductions of lifetime fatigue damage up to 17% at the blade root and up to 4% at the tower bottom. Copyright © 2016 John Wiley & Sons, Ltd.

Recently, there has been an increased focus on the immune checkpoint protein PD-1 and its ligand PD-L1 due to the discovery that blocking the PD-1/PD-L1 pathway with monoclonal antibodies elicits striking clinical results in many different malignancies. We have described naturally occurring PD-L1-specific T cells that recognize both PD-L1-expressing immune cells and malignant cells. Thus, PD-L1-specific T cells have the ability to modulate adaptive immune reactions by reacting to regulatory cells. Thus, utilization of PD-L1-derived T cell epitopes may represent an attractive vaccination strategy for targeting the tumor microenvironment and for boosting the clinical effects of additional anticancer immunotherapy. This review summarizes present information about PD-L1 as a T cell antigen, depicts the initial findings about the function of PD-L1-specific T cells in the adjustment of immune responses, and discusses future opportunities.

A T-cell permissive tumor microenvironment, characterized by the presence of activated T cells and low fibrotic activity is crucial for response to immune checkpoint inhibitors (ICIs). Granzyme B has been shown to promote T-cell migration through the basement membrane by the degradation of type IV collagen. In this study, we evaluated the biomarker potential of measuring granzyme B-mediated degradation of type IV collagen (C4G) in combination with a fibroblast activation biomarker (PRO-C3) non-invasively for identifying metastatic melanoma patients responding to the ICI ipilimumab. A monoclonal antibody was generated against C4G and used to develop a competitive electro-chemiluminescence immunoassay. C4G and PRO-C3 were measured in pretreatment serum from metastatic melanoma patients (n = 54). The C4G assay was found specific for a granzyme B-generated neo-epitope on type IV collagen. The objective response rate (ORR) was 2.6-fold higher (18% vs. 7%) in patients with high C4G levels (&gt, 25th percentile) vs. low levels (&le, 25th percentile). Likewise, high C4G levels at baseline were associated with longer overall survival (OS) (log-rank, p = 0.040, and hazard ratio (HR) = 0.48, 95%CI: 0.24&ndash, 0.98, p = 0.045). Combining high C4G with low PRO-C3 correlated with improved OS with a median OS of 796 days vs. 273 days (p = 0.0003) and an HR of 0.30 (95%CI: 0.15&ndash, 0.60, p = 0.0006). In conclusion, these results suggest that high granzyme B degraded type IV collagen (C4G) combined with low PRO-C3 quantified non-invasively has the potential to identify the responders to ICI therapy.

Background: Non-invasive biomarkers for predicting response to immune checkpoint inhibitors (ICI) are urgently needed. Crucial for response to ICI's is a T-cell permissive tumor microenvironment (hot tumor), characterized by presence of activated T-cells and low fibrotic activity. Activated T-cells release the protease granzyme B (GzB), which can cleave type IV collagen during tumor infiltration. We evaluated the potential of measuring GzB generated type IV collagen degradation fragments in a liquid biopsy for identifying metastatic melanoma (MM) patients responding to ipilimumab. Methods: A monoclonal antibody was raised against a GzB generated neo-epitope on type IV collagen (C4G), and used to develop a technically robust competitive electro-chemiluminescence immunoassay (ECLIA). C4G was measured in serum from MM patients (n=54) before initiation of ipilimumab treatment. C4G was combined with the fibrosis biomarker PRO-C3, measuring type III collagen formation. Biomarker levels were associated with objective response rate (ORR) and overall survival (OS) outcomes. Results: The C4G assay was specific for a neo-epitope on type IV collagen degradation fragments generated by GzB. The ORR was 2.6 fold higher (18% vs 7%) in patients with high C4G levels (>25th percentile) vs low levels (≤25th percentile). Likewise, high C4G levels at baseline were associated with longer OS, with a median OS of 646 days vs 290 days for low C4G levels (HR=0.48, 95%CI: 0.24-0.98, p=0.045). When combining high C4G with low PRO-C3, the HR dropped to 0.30 (95%CI: 0.15-0.60, p=0.0006), and remained significant when adjusted for the covariates age, lactate dehydrogenase levels and prior treatment (HR: 0.35, 95%CI: 0.18-0.72, p=0.004). Conclusions: A liquid biopsy measuring granzyme B degraded type IV collagen (C4G) as a surrogate of active immune infiltration into the tumor microenvironment is associated with response to the ICI therapy ipilimumab. When combining C4G with the fibrosis biomarker PRO-C3, patients with this special phenotype - low fibrosis and high immune infiltration - have an even better chance of responding compared to high C4G levels alone. If validated, this suggests that specific collagen remodeling biomarkers (C4G+PRO-C3) have potential for predicting response to ICI's in clinical cancer trials. Citation Format: Christina Jensen, Dovile Sinkeviciute, Daniel H. Madsen, Patrik Önnerfjord, Morten Hansen, Henrik Schmidt, Inge Marie Svane, Morten A. Karsdal, Nicholas Willumsen. Liquid biopsy reflecting a T-cell permissive tumor microenvironment identifies metastatic melanoma patients responding to immune checkpoint inhibitor therapy [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 3091.

Left atrial function assessed by 2D speckle tracking echocardiography has shown to predict recurrence of atrial fibrillation (AF) after catheter ablation (CA). However, no studies have used implantable loop recorders to verify recurrence. The purpose of this study was to evaluate the predictive

Various modal analysis methods are available for single-rotor wind turbines, but there is no report and guidance on the modal property analysis of multi-rotor wind turbines. This paper presents a dynamic modeling method for the modal response analysis of a wind turbine with two three-bladed isotropic rotors. The equations of motion are derived using Lagrange’s equations and are further linearized at a steady-state equilibrium. To avoid using Floquet Theory to remove the periodic coefficients, multi-blade coordinates are utilized. Comparison between the numerical simulations and a high-fidelity model in HAWC2 shows agreements in terms of modal frequencies. The results shows that the whirling modes splits into symmetric and asymmetric rotor modes.

Background The extracellular matrix (ECM) is a major component of tumors. Several recent findings link ECM composition with outcome in cancer patients treated with immune checkpoint inhibitors (ICIs). The aim here was to explore circulating ECM- and tissue-derived metabolites to enable clustering of patients with metastatic melanoma (MM) into putative endotypes and evaluate patient characteristics and outcome when treated with Ipilimumab accordingly. Methods Serum was collected from MM patients prior to treatment with Ipilimumab (n = 64). Wards hierarchical clustering of patients was based on 15 ECM- and tissue-derived metabolites measured in serum by ELISA. Identified clusters (endotypes) were compared to clinical characteristics and evaluated for associations with disease control rate (DCR), progression free survival (PFS) and overall survival (OS). Results Three putative endotypes (cluster A, B, C) were identified including 14, 30 and 20 patients, respectively. Overall a stepwise increase in ECM metabolite median levels were detected from C-B-A with the largest absolute change seen from B-A. There was no difference between A, B, and C according to age, gender, lactate dehydrogenase levels, number of metastasized organs and whether patients were treated previously. At follow-up, the DCR was 0%, 60% and 45% in A, B, and C, respectively. Likewise, patients with endotype A had a median PFS/OS time of 69/85 days versus 174/520 and 165/589 days for endotype B and C. In support, endotype A predicted for poor survival outcomes (PFSAvsB+C:HR=3.9, 95%CI:2.0-7.6, p = 0.0001; OSAvsB+C:HR:2.5, 95%CI:1.2-4.9, p = 0.0108). Conclusions Hierarchical clustering of MM patients based on 15 ECM- and tissue-derived metabolites measured in a liquid biopsy identifies 3 putative endotypes. One endotype (A) seems to reflect patients with an overall high and differentiated ECM turnover profile. These patients experience poor outcome when treated with Ipilimumab. If validated, this supports a link between ECM remodeling and outcome in cancer patients treated with ICIs. Legal entity responsible for the study Nordic Bioscience. Funding Has not received any funding. Disclosure N. Willumsen: Full / Part-time employment: Nordic Bioscience. C.L. Bager: Full / Part-time employment: Nordic Bioscience. C. Jensen: Full / Part-time employment: Nordic Bioscience. M.A. Karsdal: Leadership role, Shareholder / Stockholder / Stock options, Full / Part-time employment: Nordic Bioscience. H. Schmidt: Advisory / Consultancy, Travel / Accommodation / Expenses: BMS; Advisory / Consultancy: Incyte; Advisory / Consultancy: Novartis; Research grant / Funding (institution): MSD; Travel / Accommodation / Expenses: Roche. I.M. Svane: Honoraria (self), Advisory / Consultancy, Travel / Accommodation / Expenses: Novartis; Honoraria (self), Advisory / Consultancy: Pierre Faber; Honoraria (self), Advisory / Consultancy, Travel / Accommodation / Expenses: MSD; Honoraria (self), Advisory / Consultancy: Incyte; Honoraria (self): Roche-Genentech; Travel / Accommodation / Expenses: Pfizer. All other authors have declared no conflicts of interest.

Short chain fatty acids (SCFAs), such as acetate, butyrate and propionate, are products of microbial macronutrients fermentation that distribute systemically and are believed to modulate host immune responses. Recent data have indicated that certain SCFAs, such as butyrate and propionate, directly modulate human dendritic cell (DC) function. Given the role of DCs in initiating and shaping the adaptive immune response, we now explore how SCFAs affect the activation of antigen-specific CD8+ T cells stimulated with autologous, MART1 peptide-pulsed DC. We show that butyrate reduces the frequency of peptide-specific CD8+ T cells and, together with propionate, inhibit the activity of those cells. On the contrary, acetate does not affect them. Importantly, butyrate and propionate inhibit the production of IL-12 and IL-23 in the DCs and exogenous IL-12 fully restores the activation of the MART-1-specific CD8+ T cells, whereas IL-23 has no effect. In conclusion, these results point to a pivotal role of butyrate and propionate in modulating CD8+ T cell activation via the inhibition of IL-12 secretion from DCs. These findings reveal a novel mechanism whereby bacterial fermentation products may modulate CD8+ T cell function with possible implications in anti-cancer immunotherapy.

Wind turbine multidisciplinary design optimization is currently the focus of several investigations because it has showed potential in reducing the cost of energy. This design approach requires fast methods to evaluate wind turbine loads with a sufficiently high level of fidelity. This paper presents a method to estimate wind turbine fatigue damage suited for optimization design applications. The method utilizes a high-order linear wind turbine model. The model comprehends a detailed description of the wind turbine and the controller. The fatigue is computed with a spectral method applied to power spectral densities of wind turbine sensor responses to turbulent wind. In this paper, the model is validated both in time domain and frequency domain with a nonlinear aeroservoelastic model. The approach is compared quantitatively against fatigue damage obtained from the power spectra of time series evaluated with nonlinear aeroservoelastic simulations and qualitatively against rainflow counting. Results are presented for three cases: load evaluation at normal operation in the full wind speed range, load change evaluation due to two different controller tunings at normal operation at three different wind speeds above rated and load dependency on the number of turbulence seeds used for their evaluation. For the full-range normal operation, the maximum difference between the two frequency domain-based estimates of the tower base lateral fatigue moments is 36%, whereas the differences for the other sensors are less than 15%. For the load variation evaluation, the maximum difference of the tower base longitudinal bending moment variation is 22%. Such large difference occurs only when the change in controller tuning has a low effect on the loads. Furthermore, results show that loads evaluated with the presented method are less dependent on the turbulent wind realization; therefore, less turbulence seeds are required compared with time-domain simulations to remove the dependency on the wind realization used to estimate loads. Copyright © 2015 John Wiley & Sons, Ltd.

Personalized cancer immunotherapy based on infusion of T cells holds the promise to specifically target a patient’s individual tumor. Accumulating evidence indicates that the T cells mediating these tumor regressions after cancer immunotherapies may primarily target patient-specific mutations expressed by the patients’ tumors and that the presence of these “neo-antigen” specific T-cells may be related to a high number of mutations in the tumor. In melanoma, treatment with autologous tumor-infiltrating lymphocytes (TILs) can mediate durable complete responses. Previous trials investigating TIL therapy in solid tumors other than melanoma have shown limited success, however none of these early trials used current preparative chemotherapy regimens, and the methods for in vitro lymphocyte expansion have changed considerably. New advances and understandings in T cell based immunotherapies have stimulated the interest in developing this approach for other indications. Here, we summarize the early clinical data in the field of adoptive cell transfer therapy (ACT) using tumor-infiltrating lymphocytes for patients with renal cell carcinoma (RCC) and ovarian cancer (OC). In addition we describe the major advances in the characterization and application of TIL therapy for patients with RCC and OC.

The enzyme arginase-1 reduces the availability of arginine to tumor-infiltrating immune cells, thus reducing T-cell functionality in the tumor milieu. Arginase-1 is expressed by some cancer cells and by immune inhibitory cells, such as myeloid-derived suppressor cells (MDSCs) and tumor-associated macrophages (TAMs), and its expression is associated with poor prognosis. In the present study, we divided the arginase-1 protein sequence into overlapping 20-amino-acid-long peptides, generating a library of 31 peptides covering the whole arginase-1 sequence. Reactivity towards this peptide library was examined in PBMCs from cancer patients and healthy individuals. IFNγ ELISPOT revealed frequent immune responses against multiple arginase-1-derived peptides. We further identified a hot-spot region within the arginase-1 protein sequence containing multiple epitopes recognized by T cells. Next, we examined in vitro-expanded tumor-infiltrating lymphocytes (TILs) isolated from melanoma patients, and detected arginase-1-specific T cells that reacted against epitopes from the hot-spot region. Arginase-1-specific CD4+T cells could be isolated and expanded from peripheral T cell pool of a patient with melanoma, and further demonstrated the specificity and reactivity of these T cells. Overall, we showed that arginase-1-specific T cells were capable of recognizing arginase-1-expressing cells. The activation of arginase-1-specific T cells by vaccination is an attractive approach to target arginase-1-expressing malignant cells and inhibitory immune cells. In the clinical setting, the induction of arginase-1-specific immune responses could induce or increase Th1 inflammation at the sites of tumors that are otherwise excluded due to infiltration with MDSCs and TAMs.

BACKGROUND. Metformin reduces plasma glucose and has been shown to increase glucagon-like peptide 1 (GLP-1) secretion. Whether this is a direct action of metformin on GLP-1 release, and whether some of the glucose-lowering effect of metformin occurs due to GLP-1 release, is unknown. The current study investigated metformin-induced GLP-1 secretion and its contribution to the overall glucose-lowering effect of metformin and underlying mechanisms in patients with type 2 diabetes. METHODS. Twelve patients with type 2 diabetes were included in this placebo-controlled, double-blinded study. On 4 separate days, the patients received metformin (1,500 mg) or placebo suspended in a liquid meal, with subsequent i.v. infusion of the GLP-1 receptor antagonist exendin9-39 (Ex9-39) or saline. During 240 minutes, blood was sampled. The direct effect of metformin on GLP-1 secretion was tested ex vivo in human ileal and colonic tissue with and without dorsomorphin-induced inhibiting of the AMPK activity. RESULTS. Metformin increased postprandial GLP-1 secretion compared with placebo (P = 0.014), and the postprandial glucose excursions were significantly smaller after metformin + saline compared with metformin + Ex9-39 (P = 0.004). Ex vivo metformin acutely increased GLP-1 secretion (colonic tissue, P < 0.01; ileal tissue, P < 0.05), but the effect was abolished by inhibition of AMPK activity. CONCLUSIONS. Metformin has a direct and AMPK-dependent effect on GLP-1–secreting L cells and increases postprandial GLP-1 secretion, which seems to contribute to metformin’s glucose-lowering effect and mode of action. TRIAL REGISTRATION. {"type":"clinical-trial","attrs":{"text":"NCT02050074","term_id":"NCT02050074"}}NCT02050074 (https://clinicaltrials.gov/ct2/show/{"type":"clinical-trial","attrs":{"text":"NCT02050074","term_id":"NCT02050074"}}NCT02050074). FUNDING. This study received grants from the A.P. Moller Foundation, the Novo Nordisk Foundation, the Danish Medical Association research grant, the Australian Research Council, the National Health and Medical Research Council, and Pfizer Inc.

Immune checkpoint blockers (ICB) have become pivotal therapies in the clinical armamentarium against metastatic melanoma (MMel). Given the frequency of immune related adverse events and increasing use of ICB, predictors of response to CTLA-4 and/or PD-1 blockade represent unmet clinical needs. Using a systems biology-based approach to an assessment of 779 paired blood and tumor markers in 37 stage III MMel patients, we analyzed association between blood immune parameters and the functional immune reactivity of tumor-infiltrating cells after ex vivo exposure to ICB. Based on this assay, we retrospectively observed, in eight cohorts enrolling 190 MMel patients treated with ipilimumab, that PD-L1 expression on peripheral T cells was prognostic on overall and progression-free survival. Moreover, detectable CD137 on circulating CD8+ T cells was associated with the disease-free status of resected stage III MMel patients after adjuvant ipilimumab + nivolumab (but not nivolumab alone). These biomarkers should be validated in prospective trials in MMel., The clinical management of metastatic melanoma requires predictors of the response to checkpoint blockade. Here, the authors use immunological assays to identify potential prognostic/predictive biomarkers in circulating blood cells and in tumor-infiltrating lymphocytes from patients with resected stage III melanoma.

In this paper we introduce an optimized Markov Chain Monte Carlo (MCMC) technique for solving the integer least-squares (ILS) problems, which include Maximum Likelihood (ML) detection in Multiple-Input Multiple-Output (MIMO) systems. Two factors contribute to the speed of finding the optimal solution by the MCMC detector: the probability of the optimal solution in the stationary distribution, and the mixing time of the MCMC detector. Firstly, we compute the optimal value of the "temperature" parameter, in the sense that the temperature has the desirable property that once the Markov chain has mixed to its stationary distribution, there is polynomially small probability ($1/\mbox{poly}(N)$, instead of exponentially small) of encountering the optimal solution. This temperature is shown to be at most $O(\sqrt{SNR}/\ln(N))$, where $SNR$ is the signal-to-noise ratio, and $N$ is the problem dimension. Secondly, we study the mixing time of the underlying Markov chain of the proposed MCMC detector. We find that, the mixing time of MCMC is closely related to whether there is a local minimum in the lattice structures of ILS problems. For some lattices without local minima, the mixing time of the Markov chain is independent of $SNR$, and grows polynomially in the problem dimension; for lattices with local minima, the mixing time grows unboundedly as $SNR$ grows, when the temperature is set, as in conventional wisdom, to be the standard deviation of noises. Our results suggest that, to ensure fast mixing for a fixed dimension $N$, the temperature for MCMC should instead be set as $\Omega(\sqrt{SNR})$ in general. Simulation results show that the optimized MCMC detector efficiently achieves approximately ML detection in MIMO systems having a huge number of transmit and receive dimensions., Comment: 14 pages. arXiv admin note: substantial text overlap with arXiv:1203.2213

Bile acid sequestrants have been used for decades for the treatment of hypercholesterolaemia. Sequestering of bile acids in the intestinal lumen interrupts enterohepatic recirculation of bile acids, which initiate feedback mechanisms on the conversion of cholesterol into bile acids in the liver, thereby lowering cholesterol concentrations in the circulation. In the early 1990s, it was observed that bile acid sequestrants improved glycaemic control in patients with type 2 diabetes. Subsequently, several studies confirmed the finding and recently – despite elusive mechanisms of action – bile acid sequestrants have been approved in the USA for the treatment of type 2 diabetes. Nowadays, bile acids are no longer labelled as simple detergents necessary for lipid digestion and absorption, but are increasingly recognised as metabolic regulators. They are potent hormones, work as signalling molecules on nuclear receptors and G protein-coupled receptors and trigger a myriad of signalling pathways in many target organs. The most described and well-known receptors activated by bile acids are the farnesoid X receptor (nuclear receptor) and the G protein-coupled cell membrane receptor TGR5. Besides controlling bile acid metabolism, these receptors are implicated in lipid, glucose and energy metabolism. Interestingly, activation of TGR5 on enteroendocrine L cells has been suggested to affect secretion of incretin hormones, particularly glucagon-like peptide 1 (GLP1 (GCG)). This review discusses the role of bile acid sequestrants in the treatment of type 2 diabetes, the possible mechanism of action and the role of bile acid-induced secretion of GLP1 via activation of TGR5.

One of the primary criteria for extracting energy from the wind using horizontal axis upwind wind turbines is the ability to align the rotor axis with the dominating wind direction. The conventional way of estimating the direction of the incoming flow is by using transducers placed atop the nacelle and downwind of the rotor. Recent studies have suggested methods based on advanced upwind measurement technologies for estimating the inflow direction and improving the yaw alignment. In this study, the potential of increased power output with improved yaw alignment is investigated by assessing the performance of a current measurement and yaw control system. The performance is assessed by analyzing data containing upwind wind speed and direction measurements from a met mast, and yaw angle and power production measurements from an operating offshore wind turbine. The results of the analysis indicate that the turbine is operating with a wind speed-dependent yaw error distribution. The theoretical annual energy production loss due to the yaw error distribution of the existing system is estimated to approximately 0.2%. Copyright © 2014 John Wiley & Sons, Ltd.

The CCR7 ligands CCL19 and CCL21 are increasingly recognized as functionally different (biased). Using mature human dendritic cells (DCs), we show that CCL19 is more potent than CCL21 in inducing 3D chemotaxis. Intriguingly, CCL21 induces prolonged and more efficient ERK1/2 activation compared with CCL19 and a C-terminal truncated (tailless) CCL21 in DCs. In contrast, tailless-CCL21 displays increased potency in DC chemotaxis compared with native CCL21. Using a CCL21-specific antibody, we show that CCL21, but not tailless-CCL21, accumulates at the cell surface. In addition, removal of sialic acid from the cell surface by neuraminidase treatment impairs ERK1/2 activation by CCL21, but not by CCL19 or tailless-CCL21. Using standard laboratory cell lines, we observe low potency of both CCL21 and tailless-CCL21 in G protein activation and β-arrestin recruitment compared with CCL19, indicating that the tail itself does not improve receptor interaction. Chemokines interact with their receptors in a stepwise manner with ultimate docking of their N-terminus into the main binding pocket. Employing site-directed mutagenesis we identify residues in this pocket of selective CCL21 importance. We also identify a molecular switch in the top of TM7 important for keeping CCR7 in an inactive conformation (Tyr312), as introduction of the chemokine receptor-conserved Glu (or Ala) induces high constitutive activity. Summarized, we show that the interaction of the tail of CCL21 with polysialic acid is needed for strong ERK signaling, whereas it impairs CCL21-mediated chemotaxis and has no impact on receptor docking consistent with the current model of chemokine:receptor interaction. This indicates that future selective pharmacological targeting of CCL19 versus CCL21 should focus on a differential targeting of the main receptor pocket, while selective targeting of tailless-CCL21 versus CCL21 and CCL19 requires targeting of the glycosaminoglycan (GAG) interaction.

e14050 Background: The extracellular matrix (ECM) is a major component of tumors. Several recent findings link ECM composition with outcome in cancer patients treated with immune checkpoint inhibitors (ICIs). The aim here was to explore circulating ECM- and tissue-derived metabolites to enable clustering of patients with metastatic melanoma (MM) into putative endotypes and evaluate patient characteristics and outcome when treated with Ipilimumab accordingly. Methods: Serum was collected from MM patients prior to treatment with Ipilimumab (n = 64). Wards hierarchical clustering of patients was based on 15 ECM- and tissue-derived metabolites measured in serum by ELISA. Identified clusters (endotypes) were compared to clinical characteristics and evaluated for associations with disease control rate (DCR), progression free survival (PFS) and overall survival (OS). Results: Three putative endotypes (cluster A, B, C) were identified including 14, 30 and 20 patients, respectively. Overall a stepwise increase in ECM metabolite median levels were detected from C-B-A with the largest absolute change seen from B-A. There was no difference between A, B, and C according to age, gender, lactate dehydrogenase levels, number of metastasized organs and whether patients were treated previously. At follow-up, the DCR was 0%, 60% and 45% in A, B, and C, respectively. Likewise, patients with endotype A had a median PFS/OS time of 69/85 days versus 174/520 and 165/589 days for endotype B and C. In support, endotype A predicted for poor survival outcomes (PFSAvsB+C:HR = 3.9, 95%CI:2.0-7.6, p = 0.0001; OSAvsB+C:HR:2.5, 95%CI:1.2-4.9, p = 0.0108). Conclusions: Hierarchical clustering of MM patients based on 15 ECM- and tissue-derived metabolites measured in a liquid biopsy identifies 3 putative endotypes. One endotype (A) seems to reflect patients with an overall high and differentiated ECM turnover profile. These patients experience poor outcome when treated with Ipilimumab. If validated, this supports a link between ECM remodeling and outcome in cancer patients treated with ICIs.

The intimate balance between peripheral tolerance and adaptive immunity has profound implications in several disease settings. Interleukin-12 (IL-12) plays a major role in immunity to intracellular pathogens and cancer by controlling IFNγ-dependent adaptive immunity. The transient production of the bioactive IL-12p70 heterodimer and the concurrent expression of interleukin-10 (IL-10) serves as a myeloid checkpoint to avoid immunopathology. Here, long-term exposure to inflammatory stimuli was evaluated on monocyte-derived dendritic cells (DCs) activated with lipopolysaccharide (LPS) and gamma interferon (IFNγ). The secretion of IFNγ from co-cultures with allogeneic T-cells present in peripheral blood mononuclear cells from healthy volunteers served as a measure of T-cell activation.The secretion of IFNγ from co-cultures was progressively lost as exposure of DCs to inflammatory stimuli was extended from one up to four days prior to co-culture or following IL-12p70 antibody-mediated blockade. Most pronounced was the 12-fold reduction (N = 9 donor pairs) seen with four-day activated DCs. Furthermore, at four days of activation, a significant fraction of DCs underwent apoptosis concomitant with their increased release of IL-10 and a striking 10-fold drop in levels of IL-12p70 as compared with DCs activated one, two or three days. Furthermore, after four days of activation, DC-derived IL-12p70 was inversely correlated with IL-10 and with IFNγ derived from co-cultures. It is currently an open question whether IL-12p70 naturally degrades after four days of activation or whether apoptotic DCs actively stimulate the degradation of IL-12p70. Citation Format: Morten Hansen, Laura Stentoft Carstensen, Andreas Obers, Inge Marie Stentoft Svane. Secreted IL-12p70 from long-term activated dendritic cells is lost concomitant with their apoptosis and release of IL-10 [abstract]. In: Proceedings of the Fourth CRI-CIMT-EATI-AACR International Cancer Immunotherapy Conference: Translating Science into Survival; Sept 30-Oct 3, 2018; New York, NY. Philadelphia (PA): AACR; Cancer Immunol Res 2019;7(2 Suppl):Abstract nr A079.

Wind turbine controllers are commonly designed on the basis of low-order linear models to capture the aeroelastic wind turbine response due to control actions and disturbances. This paper characterizes the aeroelastic wind turbine dynamics that influence the open-loop frequency response from generator torque and collective pitch control actions of a modern non-floating wind turbine based on a high-order linear model. The model is a linearization of a geometrically non-linear finite beam element model coupled with an unsteady blade element momentum model of aerodynamic forces including effects of shed vorticity and dynamic stall. The main findings are that the lowest collective flap modes have limited influence on the response from generator torque to generator speed, due to large aerodynamic damping. The transfer function from collective pitch to generator speed is affected by two non-minimum phase zeros below the frequency of the first drivetrain mode. To correctly predict the non-minimum phase zeros, it is essential to include lateral tower and blade flap degrees of freedom. Copyright © 2013 John Wiley & Sons, Ltd.

We compare measurements of the Brownian relaxation response of magnetic nanobeads in suspension using planar Hall effect sensors of cross geometry and a newly proposed bridge geometry. We find that the bridge sensor yields six times as large signals as the cross sensor, which results in a more accurate determination of the hydrodynamic size of the magnetic nanobeads. Finally, the bridge sensor has successfully been used to measure the change in dynamic magnetic response when rolling circle amplified DNA molecules are bound to the magnetic nanobeads. The change is validated by measurements performed in a commercial AC susceptometer. The presented bridge sensor is, thus, a promising component in future lab-on-a-chip biosensors for detection of clinically relevant analytes, including bacterial genomic DNA and proteins.

The nuclear factor κ light chain enhancer of activated B cells (NFκB) is constitutively active in most cancers, controlling multiple cellular processes including proliferation, invasion and resistance to therapy. NFκB is primarily regulated through the association with inhibitory proteins that are known as inhibitors of NFκB (IκBs). Increased NFκB activity in tumor cells has been correlated with decrease stability of IκB proteins, in particular IκBα. In responso to a large number of stimuli, IκB proteins are degraded by the proteasome. Cytotoxic T lymphocytes (CTLs) recognize HLA-restricted antigenic peptides that are generated by proteasomal degradation in target cells. In the present study, we demonstrate the presence of naturally occurring IκBα -specific T cells in the peripheral blood of patients suffering from several unrelated tumor types, i.e., breast cancer, malignant melanoma and renal cell carcinoma, but not of healthy controls. Furthermore, we show that such IBα-specific T cells are granzyme B-releasing, cytotoxic cells. Hence, the increased proteasomal degradation of IκBα in cancer induces IκBα-specific CTLs.

LIDAR-assisted collective pitch control shows promising results for load reduction in the full load operating region of horizontal axis wind turbines (WT). Utilizing LIDARs in WT control can be approached in different ways; One method is to design the WT controller from ground up based on the LIDAR measurements. Nevertheless, to make the LIDAR-assisted controller easily implementable on existing wind turbines, one can design a controller that is added to the original and existing WT controller. This add-on solution makes it easier to prove the applicability and performance of the LIDAR-assisted WT control and opens the market of retrofitting existing wind turbines with the new technology. In this paper, we suggest a model predictive controller (MPC) that is added to the basic gain scheduled PI controller of a WT to enhance the performance of the closed loop system using LIDAR measurements. The performance of the MPC controller is compared against two controllers. The controllers are 1) a gain scheduled PI controller and 2) a controller with the same feedback as controller no. 1 and an added feed-forward loop (FF+PI controller). Simulations are used to compare their performances. The simulation scenarios include the extreme operating gust and normal power production using stochastic wind field in the full load region. The results show superior performance compared to the PI controller and a performance marginally better compared to the FF+PI controller. The reason for a better performance against the PI controller is that the MPC controller employs the LIDAR wind speed measurements to predict and compensate future disturbances. The MPC controller is designed based on the closed loop model of the wind turbine including the pitch actuator and therefore an appropriate pitch signal is calculated, while the FF+PI controller employs filter and delay compensation to take the actuator dynamics into account.

Tumor cells and tumor-infiltrating macrophages produce the chemokine CCL22, which attracts regulatory T cells (Tregs) into the tumor microenvironment, decreasing anticancer immunity. Here, we investigated the possibility of targeting CCL22-expressing cells by activating specific T cells. We analyzed the CCL22 protein signal sequence, identifying a human leukocyte antigen A2- (HLA-A2-) restricted peptide epitope, which we then used to stimulate peripheral blood mononuclear cells (PMBCs) to expand populations of CCL22-specific T cells in vitro. T cells recognizing an epitope derived from the signal-peptide of CCL22 will recognize CCL22-expressing cells even though CCL22 is secreted out of the cell. CCL22-specific T cells recognized and killed CCL22-expressing cancer cells. Furthermore, CCL22-specific T cells lysed acute monocytic leukemia cells in a CCL22 expression-dependent manner. Using the Enzyme-Linked ImmunoSPOT assay, we examined peripheral blood mononuclear cells from HLA-A2+ cancer patients and healthy volunteers for reactivity against the CCL22-derived T-cell epitope. This revealed spontaneous T-cell responses against the CCL22-derived epitope in cancer patients and in healthy donors. Finally, we performed tetramer enrichment/depletion experiments to examine the impact of HLA-A2-restricted CCL22-specific T cells on CCL22 levels among PMBCs. The addition or activation of CCL22-specific T cells decreased the CCL22 level in the microenvironment. Activating CCL22-specific T cells (e.g., by vaccination) may directly target cancer cells and tumor-associated macrophages, thereby modulating Treg recruitment into the tumor environment and augmenting anticancer immunity.

Background aims Maturation of dendritic cells (DCs) induces their homing from peripheral to lymphatic tissues guided by CCL21. However, in vitro matured human monocyte-derived DC cancer vaccines injected intradermally migrate poorly to lymph nodes (LNs). In vitro maturation protocols generate DCs with high (type 1 DCs) or low (prostaglandin E 2 [PGE 2 ]-DCs) autocrine CCL19 levels, which may potentially interfere with LN homing of DCs. Methods. Employing a three-dimensional (3D) chemotaxis assay, chemokine competition/desensitization studies and short interfering RNA (siRNA) against CCL19, we analyzed the effect of autocrine CCL19 on in vitro migration of human DCs toward CCL21. Results. Using human monocyte-derived DCs in a 3D chemotaxis assay, we are the first to demonstrate that CCL19 more potently induces directed migration of human DCs compared with CCL21. When comparing migration of type 1 DCs and PGE 2 -DCs, migration of type 1 DCs was strikingly impaired compared with PGE 2 -DCs, but only toward low concentrations of CCL21. When type 1 DCs were cultured overnight in fresh culture medium (reducing autocrine CCL19 levels), a rescuing effect was observed on migration toward low concentrations of CCL21 in a 3D chemotaxis assay. Finally pre-incubation with CCL19 negatively affected PGE 2 -DC migration, whereas silencing of CCL19 by siRNA improved type 1 DC migration. Importantly, in both cases, the effect was observed only at low concentrations of CCL21. Conclusions. Our results demonstrate that autocrine CCL19 negatively affects DC migratory potential toward CCL21, the potency difference between CCL19 and CCL21 being the underlying cause. CCL19 secretion level of in vitro matured DCs is an important indicator of DC vaccine homing potential.

The modal dynamics of structures with bladed isotropic rotors is analyzed using Hill's method. First, analytical derivation of the periodic system matrix shows that isotropic rotors with more than two blades can be represented by an exact Fourier series with 3/rev as the highest order. For 2-bladed rotors, the inverse mass matrix has an infinite Fourier series with harmonic components of decreasing norm, thus the system matrix can be approximated by a truncated Fourier series of predictable accuracy. Second, a novel method for automatically identifying the principal solutions of Hill's eigenvalue problem is introduced. The corresponding periodic eigenvectors can be used to compute symmetric and anti-symmetric components of the 2-bladed rotor motion, and the additional forward and backward whirling components for rotors with more than two blades. Finally, the generic methods are used on a simple wind turbine model consisting of three degrees of freedom for each blade and seven degrees of freedom for the nacelle and drivetrain. The modal dynamics of a 3-bladed 10MW turbine from previous studies is recaptured. Removing one blade, the larger and higher harmonic terms in the system matrix lead to resonant modal couplings for the 2-bladed turbine that do not exist for the 3-bladed turbine, and that excitation of a single mode of a 2-bladed turbine leads to responses at several frequencies in both the ground-fixed and rotating blade frames of reference which complicates the interpretation of simulated or measured turbine responses.

An investigation of a control strategy to allow wind turbines to operate at high wind speeds by derating the rotor speed and generator torque set-points is presented. The investigation analyzes the wind turbine aeroservoelastic behavior in the above rated operational range by computing the aerodynamic gains and closed-loop eigenvalue solutions using a high-fidelity linear model. A simple strategy to reduce the reference rotor speed based on a pitch angle feedback is presented and analyzed. It is shown that high aerodynamic gains for operation at high wind speeds requires special handling in the scheduling of the controller gains. The computed closed-loop modal frequencies and damping ratios show how most turbine modes become less damped as the rotor speed is derated, and at very high winds the frequency and damping of the first drivetrain torsion mode are significantly reduced. Possible resonance problems can also be seen from the computed frequencies, and these problems may be worsened by the decreased damping during storm-ride-through. Finally it is shown that the dynamics of the pitch feedback to the derated generator speed is significantly affected by the operational wind speed, resulting in a slow response at high wind speeds.