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The automation of online order retrieval for Chemical Abstracts Service Document Delivery Service was accomplished by shifting to an order retrieval/dispatch process linked to a Unix network. The Unix-based environment, its terminal emulation, page-break, and user-friendly interface software, and later enhancements are reviewed. Resultant increase in processing productivity is discussed. (MBR)

Brain metastasis (BrM) is a common malignancy, predominantly originating from lung, melanoma, and breast cancers. The vasculature is a key component of the BrM tumor microenvironment with critical roles in regulating metastatic seeding and progression. However, the heterogeneity of the major BrM vascular components, namely endothelial and mural cells, is still poorly understood. We perform single-cell and bulk RNA-sequencing of sorted vascular cell types and detect multiple subtypes enriched specifically in BrM compared to non-tumor brain, including previously unrecognized immune regulatory subtypes. We integrate the human data with mouse models, creating a platform to interrogate vascular targets for the treatment of BrM. We find that the CD276 immune checkpoint molecule is significantly upregulated in the BrM vasculature, and anti-CD276 blocking antibodies prolonged survival in preclinical trials. This study provides important insights into the complex interactions between the vasculature, immune cells, and cancer cells, with translational relevance for designing therapeutic interventions., Competing Interests: Declaration of interests M.E.H. has an advisory role at TME Pharma., (Copyright © 2023 The Author(s). Published by Elsevier Inc. All rights reserved.)

Melanoma frequently metastasises to the brain, and a detailed understanding of the molecular and cellular mechanisms underlying melanoma cell extravasation across the blood-brain barrier (BBB) is important for preventing brain metastasis formation. Making use of primary mouse brain microvascular endothelial cells (pMBMECs) as an in vitro BBB model, we imaged the interaction of melanoma cells into pMBMEC monolayers. We observed exclusive junctional intercalation of melanoma cells and confirmed that melanoma-induced pMBMEC barrier disruption can be rescued by protease inhibition. Interleukin (IL)-1β stimulated pMBMECs or PECAM-1-knockout (-ko) pMBMECs were employed to model compromised BBB barrier properties in vitro and to determine increased melanoma cell intercalation compared to pMBMECs with intact junctions. The newly generated brain-homing melanoma cell line YUMM1.1-BrM4 was used to reveal increased in vivo extravasation of melanoma cells across the BBB of barrier-compromised PECAM-1-deficient mice compared to controls. Taken together, our data indicate that preserving BBB integrity is an important measure to limit the formation of melanoma-brain metastasis.

Neutrophils are abundant immune cells in the circulation and frequently infiltrate tumors in substantial numbers. However, their precise functions in different cancer types remain incompletely understood, including in the brain microenvironment. We therefore investigated neutrophils in tumor tissue of glioma and brain metastasis patients, with matched peripheral blood, and herein describe the first in-depth analysis of neutrophil phenotypes and functions in these tissues. Orthogonal profiling strategies in humans and mice revealed that brain tumor-associated neutrophils (TANs) differ significantly from blood neutrophils and have a prolonged lifespan and immune-suppressive and pro-angiogenic capacity. TANs exhibit a distinct inflammatory signature, driven by a combination of soluble inflammatory mediators including tumor necrosis factor alpha (TNF-ɑ) and Ceruloplasmin, which is more pronounced in TANs from brain metastasis versus glioma. Myeloid cells, including tumor-associated macrophages, emerge at the core of this network of pro-inflammatory mediators, supporting the concept of a critical myeloid niche regulating overall immune suppression in human brain tumors., Competing Interests: Declaration of interests D.C. has received consulting fees from Seed Biosciences S.A.; P.W. has provided consulting for Almax, Bayer, Sanofi, and Genentech. A.H. is a paid consultor for Flagship Pioneering, Inc. for matters unrelated to this study; M.E.H. has an advisory role at TME Pharma; J.A.J. received an honorarium for speaking at a Bristol Meyers Squibb research symposium and previously served on the scientific advisory board of Pionyr Immunotherapeutics., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

The immune-specialized environment of the healthy brain is tightly regulated to prevent excessive neuroinflammation. However, after cancer development, a tissue-specific conflict between brain-preserving immune suppression and tumor-directed immune activation may ensue. To interrogate potential roles of T cells in this process, we profiled these cells from individuals with primary or metastatic brain cancers via integrated analyses on the single-cell and bulk population levels. Our analysis revealed similarities and differences in T cell biology between individuals, with the most pronounced differences observed in a subgroup of individuals with brain metastasis, characterized by accumulation of CXCL13-expressing CD39 + potentially tumor-reactive T (pTRT) cells. In this subgroup, high pTRT cell abundance was comparable to that in primary lung cancer, whereas all other brain tumors had low levels, similar to primary breast cancer. These findings indicate that T cell-mediated tumor reactivity can occur in certain brain metastases and may inform stratification for treatment with immunotherapy., (© 2023. The Author(s).)

Brain metastases (BrMs) are the most common form of brain tumors in adults and frequently originate from lung and breast primary cancers. BrMs are associated with high mortality, emphasizing the need for more effective therapies. Genetic profiling of primary tumors is increasingly used as part of the effort to guide targeted therapies against BrMs, and immune-based strategies for the treatment of metastatic cancer are gaining momentum. However, the tumor immune microenvironment (TIME) of BrM is extremely heterogeneous, and whether specific genetic profiles are associated with distinct immune states remains unknown. Here, we perform an extensive characterization of the immunogenomic landscape of human BrMs by combining whole-exome/whole-genome sequencing, RNA sequencing of immune cell populations, flow cytometry, immunofluorescence staining, and tissue imaging analyses. This revealed unique TIME phenotypes in genetically distinct lung- and breast-BrMs, thereby enabling the development of personalized immunotherapies tailored by the genetic makeup of the tumors., Competing Interests: Declaration of interests F.K. is currently an employee at Sophia Genetics; V.Z. is a consultant for Cellestia Biotech; S.B. has received honoraria from Eli Lilly (Advisory Board) and research funding to the institution from Roche and Basilea (last 3 years); M.E.H. has an advisory role at TME Pharma; J.A.J. has received honoraria for speaking at research symposia organized by Bristol Meyers Squibb and Glenmark Pharmaceuticals (last 3 years) and currently serves on the scientific advisory board of Pionyr Immunotherapeutics., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Structured catalytic membranes with high porosity and a low pressure drop are particularly suitable for industrial processes carried out at high space velocities. One of these processes is the catalytic total oxidation of volatile organic compounds, which is an economically feasible and environmentally friendly way of emission abatement. Noble metal catalysts are typically preferred due to high activity and stability. In this paper, the preparation of a thermally stable polybenzimidazole electrospun membrane, which can be used as a support for a platinum catalyst applicable in the total oxidation of volatile organic compounds, is reported for the first time. In contrast to commercial pelletized catalysts, high porosity of the membrane allowed for easy accessibility of the platinum active sites to the reactants and the catalytic bed exhibited a low pressure drop. We have shown that the preparation conditions can be tuned in order to obtain catalysts with a desired platinum particle size. In the gas-phase oxidation of ethanol, acetone, and toluene, the catalysts with Pt particle sizes 2.1 nm and 26 nm exhibited a lower catalytic activity than that with a Pt particle size of 12 nm. Catalysts with a Pt particle size of 2.1 nm and 12 nm were prepared by equilibrium adsorption, and the higher catalytic activity of the latter catalyst was ascribed to more reactive adsorbed oxygen species on larger Pt nanoparticles. On the other hand, the catalyst with a Pt particle size of 26 nm was prepared by a solvent evaporation method and contained less active polycrystalline platinum. Last but not least, the catalyst containing only 0.08 wt.% of platinum achieved high conversion (90%) of all the model volatile organic compounds at moderate temperatures (lower than 335 °C), which is important for reducing the costs of the abatement technology.

In recent years, porous materials have been extensively studied by the scientific community owing to their excellent properties and potential use in many different areas, such as gas separation and adsorption. Hyper-crosslinked porous polymers (HCLPs) have gained attention because of their high surface area and porosity, low density, high chemical and thermal stability, and excellent adsorption capabilities in comparison to other porous materials. Herein, we report the synthesis, characterization, and gas (particularly CO 2 ) adsorption performance of a series of novel styrene-based HCLPs. The materials were prepared in two steps. The first step involved radical copolymerization of divinylbenzene (DVB) and 4-vinylbenzyl chloride (VBC), a non-porous gel-type polymer, which was then modified by hyper-crosslinking, generating micropores with a high surface area of more than 700 m 2 g -1 . In the following step, the polymer was impregnated with various polyamines that reacted with residual alkyl chloride groups on the pore walls. This impregnation substantially improved the CO 2 /N 2 and CO 2 /CH 4 adsorption selectivity.

High-grade gliomas, the most common and aggressive primary brain tumors, are characterized by a complex tumor microenvironment (TME). Among the immune cells infiltrating the glioma TME, tumor-associated microglia and macrophages (TAMs) constitute the major compartment. In patients with gliomas, increased TAM abundance is associated with more aggressive disease. Alterations in TAM phenotypes and functions have been reported in preclinical models of multiple cancers during tumor development and after therapeutic interventions, including radiotherapy and molecular targeted therapies. These findings indicate that it is crucial to evaluate TAM abundance and dynamics over time. Current techniques to quantify TAMs in patients rely mainly on histological staining of tumor biopsies. Although informative, these techniques require an invasive procedure to harvest the tissue sample and typically only result in a snapshot of a small region at a single point in time. Fluorine isotope 19 MRI ( 19 F MRI) represents a powerful means to noninvasively and longitudinally monitor myeloid cells in pathological conditions by intravenously injecting perfluorocarbon-containing nanoparticles (PFC-NP). In this study, we demonstrated the feasibility and power of 19 F MRI in preclinical models of gliomagenesis, breast-to-brain metastasis, and breast cancer and showed that the major cellular source of 19 F signal consists of TAMs. Moreover, multispectral 19 F MRI with two different PFC-NP allowed us to identify spatially and temporally distinct TAM niches in radiotherapy-recurrent murine gliomas. Together, we have imaged TAMs noninvasively and longitudinally with integrated cellular, spatial, and temporal resolution, thus revealing important biological insights into the critical functions of TAMs, including in disease recurrence.

Aberrant innate immune responses to the gut microbiota are causally involved in the pathogenesis of inflammatory bowel diseases (IBD). The exact triggers and main signaling pathways activating innate immune cells and how they modulate adaptive immunity in IBD is still not completely understood. Here, we report that the PI3K/PTEN signaling pathway in dendritic cells enhances IL-6 production in a model of DSS-induced colitis. This results in exacerbated Th1 cell responses and increased mortality in DC-specific PTEN knockout (PTEN ΔDC ) animals. Depletion of the gut microbiota using antibiotics as well as blocking IL-6R signaling rescued mortality in PTEN ΔDC mice, whereas adoptive transfer of Flt3L-derived PTEN -/- DCs into WT recipients exacerbated DSS-induced colitis and increased mortality. Taken together, we show that the PI3K signaling pathway in dendritic cells contributes to disease pathology by promoting IL-6 mediated Th1 responses., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Kuttke, Hromadová, Yildirim, Brunner, Vogel, Paar, Peters, Weber, Hofmann, Kerndl, Kieler, Datler, Musiejovsky, Salzmann, Lang, Soukup, Halfmann, Sharif and Schabbauer.)

Dendritic cells (DCs) induce peripheral T cell tolerance, but cell-intrinsic signaling cascades governing their stable tolerogenesis remain poorly defined. Janus Kinase 1 (JAK1) transduces cytokine-receptor signaling, and JAK inhibitors (Jakinibs), including JAK1-specific filgotinib, break inflammatory cycles in autoimmunity. Here, we report in heterogeneous DC populations of multiple secondary lymphoid organs that JAK1 promotes peripheral T cell tolerance during experimental autoimmune encephalomyelitis (EAE). Mice harboring DC-specific JAK1 deletion exhibit elevated peripheral CD4 + T cell expansion, less regulatory T cells (Tregs), and worse EAE outcomes, whereas adoptive DC transfer ameliorates EAE pathogenesis by inducing peripheral Tregs, programmed cell death ligand 1 (PD-L1) dependently. This tolerogenic program is substantially reduced upon the transfer of JAK1-deficient DCs. DC-intrinsic IFN-γ-JAK1-STAT1 signaling induces PD-L1, which is required for DCs to convert CD4 + T cells into Tregs in vitro and attenuated upon JAK1 deficiency and filgotinib treatment. Thus, DC-intrinsic JAK1 promotes peripheral tolerance, suggesting potential unwarranted DC-mediated effects of Jakinibs in autoimmune diseases., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Human tissue samples represent an invaluable source of information for the analysis of disease-specific cellular alterations and their variation between different pathologies. In cancer research, advancing a comprehensive understanding of the unique characteristics of individual tumor types and their microenvironment is of considerable importance for clinical translation. However, investigating human brain tumor tissue is challenging due to the often-limited availability of surgical specimens. Here we describe a multimodule integrated pipeline for the processing of freshly resected human brain tumor tissue and matched blood that enables analysis of the tumor microenvironment, with a particular focus on the tumor immune microenvironment (TIME). The protocol maximizes the information yield from limited tissue and includes both the preservation of bulk tissue, which can be performed within 1 h following surgical resection, as well as tissue dissociation for an in-depth characterization of individual TIME cell populations, which typically takes several hours depending on tissue quantity and further downstream processing. We also describe integrated modules for immunofluorescent staining of sectioned tissue, bulk tissue genomic analysis and fluorescence- or magnetic-activated cell sorting of digested tissue for subsequent culture or transcriptomic analysis by RNA sequencing. Applying this pipeline, we have previously described the overall TIME landscape across different human brain malignancies, and were able to delineate disease-specific alterations of tissue-resident versus recruited macrophage populations. This protocol will enable researchers to use this pipeline to address further research questions regarding the tumor microenvironment., (© 2021. The Author(s), under exclusive licence to Springer Nature Limited.)

Pharmaceutical wastewater pollution has reached an alarming stage, as many studies have reported. Membrane separation has shown great performance in wastewater treatment, but there are some drawbacks and undesired byproducts of this process. Selective membranes could be used for pollutant investigation sensors or even for pollutant recovery. The polydimethylsiloxane (PDMS) membrane was first tested on separated and mixed antibiotic (ATB) water solutions containing sulfamethoxazole (SM), trimethoprim (TMP), and tetracycline (TET). Then, the bare and ultra-violet grafted (UV-grafted) PDMS membranes (MMA-DMAEMA 10, GMA-DMAEMA 5, and GMA-DMAEMA 10) were tested in tramadol (TRA) separation, where the diffusion coefficient was evaluated. Finally, the membranes were tested in pertraction with a mixture of SM, TMP, TET, and TRA. The membranes were characterized using the following methods: contact angle measurement, FTIR, SEM/EDX, and surface and pore analysis. The main findings were that TET was co-eluted during mixed ATB pertraction, and GMA-DMAEMA 5 was found to selectively permeate TRA over the present ATBs.

The hydrophilic and hydrophobic single-walled carbon nanotube membranes were prepared and progressively applied in sorption, filtration, and pertraction experiments with the aim of eliminating three antibiotics-tetracycline, sulfamethoxazole, and trimethoprim-as a single pollutant or as a mixture. The addition of SiO 2 to the single-walled carbon nanotubes allowed a transparent study of the influence of porosity on the separation processes. The mild oxidation, increasing hydrophilicity, and reactivity of the single-walled carbon nanotube membranes with the pollutants were suitable for the filtration and sorption process, while non-oxidized materials with a hydrophobic layer were more appropriate for pertraction. The total pore volume increased with an increasing amount of SiO 2 (from 743 to 1218 mm 3 /g) in the hydrophilic membranes. The hydrophobic layer completely covered the carbon nanotubes and SiO 2 nanoparticles and provided significantly different membrane surface interactions with the antibiotics. Single-walled carbon nanotubes adsorbed the initial amount of antibiotics in less than 5 h. A time of 2.3 s was sufficient for the filtration of 98.8% of sulfamethoxazole, 95.5% of trimethoprim, and 87.0% of tetracycline. The thicker membranes demonstrate a higher adsorption capacity. However, the pertraction was slower than filtration, leading to total elimination of antibiotics (e.g., 3 days for tetracycline). The diffusion coefficient of the antibiotics varies between 0.7-2.7 × 10 -10 , depending on the addition of SiO 2 in perfect agreement with the findings of the textural analysis and scanning electron microscopy observations. Similar to filtration, tetracycline is retained by the membranes more than sulfamethoxazole and trimethoprim.

The far-field patterns of atoms diffracted from a classical light field or from a quantum one in a photon-number state are identical. On the other hand, diffraction from a field in a coherent state, which shares many properties with classical light, displays a completely different behavior. We show that in contrast to the diffraction patterns, the interference signal of an atom interferometer with light-pulse beam splitters and mirrors in intense coherent states does approach the limit of classical fields. However, low photon numbers reveal the granular structure of light, leading to a reduced visibility since welcher-Weg (which-way) information is encoded into the field. We discuss this effect for a single photon-number state as well as a superposition of two such states.

The aim of the study was to investigate the sorption interactions between phenol and materials obtained from four different underground coal gasification (UCG) ex-situ simulations. These interactions are significant in terms of the impact of the UCG on the groundwater environment. Sorption parameters were determined for two sample types: raw coal mined from the coal-bed and then subjected to the gasification process; and char residue acquired from the cavity formed as a result of the UCG processes. Laboratory-scale tests were carried out using deionized water and aqueous solutions with increasing concentrations of phenol (from 50 mg/dm 3 to 2000 mg/dm 3 ) at 298 K. On the assumption of physical interactions (non-specific physisorption) and due to a nonlinear mass distribution of adsorbed substances as a function of equilibrium concentration, the Freundlich isotherm model was applied to describe adsorption phenomena. The isotherms have good fitting (R 2 from 0.5716 to 0.9811). Relatively high percentage phenol removal efficiency was observed for all tested chars (from 17.0% to 99.8% for the 1.0-2.5 mm fraction and from 6.9% to 99.6% for the 10.0-12.5 mm fraction). Additionally, the sorption characteristics was used to evaluate the retardation coefficients. The largest delay in the organic pollutant migration in the environment around a UCG reactor occurs for phenol transport in the layer of the post-process char from 'Wesoła' after 40 bar pressure experiment., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

In this study, we report for the first time a novel type of sorbent that can be used for mercury adsorption from the air-based off-gasses-vermiculite impregnated with alkali polysulfides and thiosulfates. In contrast to other sorbents, vermiculite exhibits superior thermal stability in air and low adsorption capacity for organic vapors. This allows for a more favorable design of the soil remediation unit-direct coupling of thermal desorber with catalytic oxidizer using air as a carrier gas. In the bench-scale test at 180 °C, the sulfur/vermiculite sorbent exhibited significantly higher efficiency for the adsorption of mercury vapor from the off-gasses than the commercial sulfur/activated carbon sorbent at its highest operating temperature (120 °C). The average mercury concentration in the adsorber off-gas decreased from 1.634 mg/m 3  for the sulfur/activated carbon to 0.008 mg/m 3 achieved with impregnated vermiculite. The total concentration of organic compounds in the soil after thermal desorption was below the detection limit of the employed analytical method.

Brain malignancies encompass a range of primary and metastatic cancers, including low-grade and high-grade gliomas and brain metastases (BrMs) originating from diverse extracranial tumors. Our understanding of the brain tumor microenvironment (TME) remains limited, and it is unknown whether it is sculpted differentially by primary versus metastatic disease. We therefore comprehensively analyzed the brain TME landscape via flow cytometry, RNA sequencing, protein arrays, culture assays, and spatial tissue characterization. This revealed disease-specific enrichment of immune cells with pronounced differences in proportional abundance of tissue-resident microglia, infiltrating monocyte-derived macrophages, neutrophils, and T cells. These integrated analyses also uncovered multifaceted immune cell activation within brain malignancies entailing converging transcriptional trajectories while maintaining disease- and cell-type-specific programs. Given the interest in developing TME-targeted therapies for brain malignancies, this comprehensive resource of the immune landscape offers insights into possible strategies to overcome tumor-supporting TME properties and instead harness the TME to fight cancer., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Glioblastoma is the most dangerous brain cancer. One reason for glioblastoma's aggressiveness are glioblastoma stem-like cells. To target them, a number of markers have been proposed (CD133, CD44, CD15, A2B5, CD36, CXCR4, IL6R, L1CAM, and ITGA6). A comprehensive study of co-expression patterns of them has, however, not been performed so far. Here, we mapped the multidimensional co-expression profile of these stemness-associated molecules. Gliomaspheres - an established model of glioblastoma stem-like cells - were used. Seven different gliomasphere systems were subjected to multicolor flow cytometry measuring the nine markers CD133, CD44, CD15, A2B5, CD36, CXCR4, IL6R, L1CAM, and ITGA6 all simultaneously based on a novel 9-marker multicolor panel developed for this study. The viSNE dimensionality reduction algorithm was applied for analysis. All gliomaspheres were found to express at least five different glioblastoma stem-like cell markers. Multi-dimensional analysis showed that all studied gliomaspheres consistently harbored a cell population positive for the molecular signature CD44+/CD133+/ITGA6+/CD36+. Glioblastoma patients with an enrichment of this combination had a significantly worse survival outcome when analyzing the two largest available The Cancer Genome Atlas datasets (MIT/Harvard Affymetrix: P = 0.0015, University of North Carolina Agilent: P = 0.0322). In sum, we detected a previously unknown marker combination - demonstrating feasibility, usefulness, and importance of high-dimensional gliomasphere marker combinatorics., (© 2018 The Authors. Journal of Cellular and Molecular Medicine published by John Wiley & Sons Ltd and Foundation for Cellular and Molecular Medicine.)

Myeloid cells, including neutrophils, are important regulators of tumor growth and metastasis. In Science, Engblom et al. (2017) reveal how lung tumors remotely engage bone-resident cells through a relay mechanism that achieves a sustained supply of tumor-promoting neutrophils., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Pancreatic ductal adenocarcinoma (PDAC) has a 5-year relative survival rate of 8% and is projected to be the second leading cause of cancer death by 2030, underscoring the urgency to develop new strategies to improve current therapeutic modalities for PDAC. Targeting pancreatic cancer stem cells (PCSCs), which are resistant to radiation and chemotherapy, is a promising strategy. A novel approach which can be readily clinically translated is to repurpose disulfiram (DSF), a drug for treating alcoholism, to target PCSCs. Chemoradiation or the combination of chemotherapy agents FOLFIRINOX, currently standard care for PDAC, can increase stemness in some established or primary PDAC cell lines. However, DSF in the presence of exogenously or endogenously supplied copper (Cu), when combined with chemotherapy or chemoradiation, targets both PCSCs and nonstem PDAC cells. Previously, we demonstrated that DSF/Cu effectively targets breast cancer stem cells in the context of fractionated radiation (FIR) by inhibiting the NF-κB-stemness gene pathway. Therefore, the hypothesis that PCSCs can be effectively targeted by incorporating DSF/Cu into the standard chemoradiation regimen consisting of 5-FU and FIR was investigated and found to be effective in vitro in targeting PCSCs, identified as either ALDH bright or CD24+/CD44+/ESA+ or sphere-forming cells, as well as nonstem PDAC cells. In vivo, the combination of IR+5-FU+DSF/Cu was more effective (72.46%) than either IR+5-FU (30.32%) or IR+FOLFIRINOX therapy (43.04%) in inhibiting growth of the mouse Panc02 tumor. These encouraging results provide a solid foundation for clinical trials to improve the outcomes of the current standard chemoradiation therapy regimen for PDAC., (Copyright © 2017 The Authors. Published by Elsevier B.V. All rights reserved.)

Donor T-cells contribute to reconstitution of protective immunity after allogeneic hematopoietic stem cell transplantation (HSCT) but must acquire specific tolerance against recipient alloantigens to avoid life-threatening graft-versus-host disease (GvHD). Systemic immunosuppressive drugs may abrogate severe GvHD, but this also impedes memory responses to invading pathogens. Here, we tested whether ex vivo blockade of CD28 co-stimulation can enable selective T-cell tolerization to alloantigens by facilitating CD80/86-cytotoxic T-lymphocyte-associated protein 4 (CTLA-4) signaling. Treatment of human allogeneic dendritic cell/T-cell co-cultures with a human CD28 blocking antibody fragment (α-huCD28) significantly abrogated subsequent allospecific immune responses, seen by decreased T-cell proliferation and of type 1 cytokine (IFN-γ and IL-2) expression. Allo-tolerization persisted after discontinuation of CD28 blockade and secondary alloantigen stimulation, as confirmed by enhanced CTLA-4 and PD-1 immune checkpoint signaling. However, T-cells retained reactivity to pathogens, supported by clonotyping of neo-primed and cross-reactive T-cells specific for Candida albicans or third-party antigens using deep sequencing analysis. In an MHC-mismatched murine model, we tolerized C57BL/6 T-cells by ex vivo exposure to a murine single chain Fv specific for CD28 (α-muCD28). Infusion of these cells, after α-muCD28 washout, into bone marrow-transplanted BALB/c mice caused allo-tolerance and did not induce GvHD-associated hepatic pathology. We conclude that selective CD28 blockade ex vivo can allow the generation of stably allo-tolerized T-cells that in turn do not induce graft-versus-host reactions while maintaining pathogen reactivity. Hence, CD28 co-stimulation blockade of donor T-cells may be a useful therapeutic approach to support the immune system after HSCT.

Maintaining dendritic cells (DC) in a state of dysfunction represents a key mechanism by which tumour cells evade recognition and elimination by the immune system. Limited knowledge about the intracellular mediators of DC dysfunction restricts success of therapies aimed at reactivating a DC-driven anti-tumour immune response. Using a cell type-specific murine knock-out model, we have identified MAPK-activated protein kinase 2 (MK2) as a major guardian of a suppressive DC phenotype in the melanoma tumour microenvironment. MK2 deletion in CD11c + cells led to an expansion of stimulatory CD103 + DCs, mounting a potent CD8 + T cell response that resulted in elimination of highly aggressive B16-F10 tumours upon toll-like receptor (TLR) activation in the presence of tumour antigen. Moreover, tumour infiltration by suppressive myeloid cells was strongly diminished. These insights into the regulation of DC functionality reveal MK2 as a targetable pathway for DC-centred immunomodulatory cancer therapies.

The PI3K signaling cascade in APCs has been recognized as an essential pathway to initiate, maintain, and resolve immune responses. In this study, we demonstrate that a cell type-specific loss of the PI3K antagonist phosphatase and tensin homolog (PTEN) in myeloid cells renders APCs toward a regulatory phenotype. APCs deficient for PTEN exhibit reduced activation of p38 MAPK and reduced expression of T cell-polarizing cytokines. Furthermore, PTEN deficiency leads to upregulation of markers for alternative activation, such as Arginase 1, with concomitant downregulation of inducible NO synthase in APCs in vitro and in vivo. As a result, T cell polarization was dysfunctional in PTEN(-/-) APCs, in particular affecting the Th17 cell subset. Intriguingly, mice with cell type-specific deletions of PTEN-targeting APCs were protected from experimental autoimmune encephalomyelitis, which was accompanied by a pronounced reduction of IL-17- and IL-22-producing autoreactive T cells and reduced CNS influx of classically activated monocytes/macrophages. These observations support the notion that activation of the PI3K signaling cascade promotes regulatory APC properties and suppresses pathogenic T cell polarization, thereby reducing the clinical symptoms and pathology of experimental autoimmune encephalomyelitis., (Copyright © 2015 by The American Association of Immunologists, Inc.)

Dendritic cell (DC)-mediated inflammation induced via TLRs is promoted by MAPK-activated protein kinase (MK)-2, a substrate of p38 MAPK. In this study we show an opposing role of MK2, by which it consolidates immune regulatory functions in DCs through modulation of p38, ERK1/2-MAPK, and STAT3 signaling. During primary TLR/p38 signaling, MK2 mediates the inhibition of p38 activation and positively cross-regulates ERK1/2 activity, leading to a reduction of IL-12 and IL-1α/β secretion. Consequently, MK2 impairs secondary autocrine IL-1α signaling in DCs, which further decreases the IL-1α/p38 but increases the anti-inflammatory IL-10/STAT3 signaling route. Therefore, the blockade of MK2 activity enables human and murine DCs to strengthen proinflammatory effector mechanisms by promoting IL-1α-mediated Th1 effector functions in vitro. Furthermore, MK2-deficient DCs trigger Th1 differentiation and Ag-specific cytotoxicity in vivo. Finally, wild-type mice immunized with LPS in the presence of an MK2 inhibitor strongly accumulate Th1 cells in their lymph nodes. These observations correlate with a severe clinical course in DC-specific MK2 knockout mice compared with wild-type littermates upon induction of experimental autoimmune encephalitis. Our data suggest that MK2 exerts a profound anti-inflammatory effect that prevents DCs from prolonging excessive Th1 effector T cell functions and autoimmunity., (Copyright © 2015 by The American Association of Immunologists, Inc.)

Purpose: This review focuses on recent advances in the field of combining radiation with immunotherapy for the treatment of malignant diseases, since various combinatorial cancer therapy approaches have lately proven highly successful., Results: With initial case reports and anecdotes progressively converting into solid clinical data, interest in cancer immunotherapy (CIT) has risen steeply. Especially immune checkpoint blockade therapies have recently celebrated tremendous successes in the treatment of severe malignancies resistant to conventional treatment strategies. Nevertheless, the high variability of patient responses to CIT remains a major hurdle, clearly indicating an urgent need for improvement. It has been suggested that successful cancer therapy most probably involves combinatorial treatment approaches. Radiotherapy (RT) has been proposed as a powerful partner for CIT due to its broad spectrum of immune modulatory characteristics. Several preclinical studies, supported by an increasing number of clinical observations, have demonstrated synergistic interactions between RT and CIT resulting in significantly improved therapy outcomes., Conclusions: Numerous reports have shown that radiation is capable of tipping the scales from tumor immune evasion to elimination in different tumor types. The next puzzle to be solved is the question of logistics - including types, schedule and dosage of combinatorial RT and CIT strategies.

Introduction: The increased level of phenylalanine (Phe) in maternal blood--hyperphenylalaninemia (mHPA) has a detrimental effect on the early development of healthy foetus (1965). The toxic effect causes spontaneous abortion or retards intrauterine growth, skeletal malformation, cardiac anomalies can appear. However the most frequent are microcephaly, mental retardation and hypotrophy., Patients and Methods: Simultaneously with the introduction of obligatory "Newborn Screening Program" in CR also the facultative screening for mHPA was introduced ("Maternal Hyperphenylalaninemia Preventive Screening Program"). Since 1975 till now 222,990 healthy pregnant women (16-47 yrs) from city Prague and its area (cca 2 mil. inh.) have been screened for increased Phe in blood by Efron's chromatographic screening test (1964); Phe cut off value: 240 micromol/l. Nonfasting venous blood has been taken in 2nd-3rd month of pregnancy during the first antenatal visit. All positive cases have been verified with quantitative Phe estimation on amino acid analyzer incl. pterines analysis in urine. For differentiation of detected mHPAs the Güttler's scheme (1980) has been used. Mutations for Phe-hydroxylase gene analyzed by restriction enzyme digestion after Guldberg (1994)., Results: The average incidence of mHPA detected at the beginning of pregnancy was found 1:8675. The major part (65.3%) of all detected mHPA belongs to mild or moderate form of phenylketonuria (PKU) with most frequent PAH gene mutations R408W, Y414C, IVS11 nt8g-a, R158Q, IVS12ntlg-a and R261Q. 19.2% corresponds to atypical or classical PKU with prevailing mutation R408W. Only in 15.3% were detected non-PKU (persistent HPA) with mutations R408W, Y414C, IVS12ntlg-a, IV11nt8g-a and A403V. 28 offsprings born from pregnancies on low-phenylalanine diet (LPD) introduced at least 2 months before the conception and during the whole pregnancy show normal psychomotoric development. In 7 offsprings without LPD or after delayed introducing or on PLD or badly monitored showed malformations (microcephaly, hypotrophy, skeletal malformations) or died., Discussion: Relatively high incidence of mHPA detected in healthy population of pregnant women of Prague area differs from findings of Buist (1989) or Levy (1994) from American pregnant women screened for mHPA from umbilical blood. We consider that screening performed at the beginning of pregnancy from nonfasting venous blood is more effective compared to umbilical blood from two reasons: the Phe level in maternal blood is increased during first trimester of pregnancy due to succing effect of placenta in comparison to decreased Phe level at the end of labour. Umbilical blood for screening of mHPA is not quite suitable to detect the atypical or mild forms of Phe disturbances which prevailed in our Slavonic population of pregnant women. (Tab. 5, Fig. 7, Ref. 16.)