Innate Adaptor Research Articles

Status of the Deep Learning-Based Shattered Pellet Injection Shard Tracking at ASDEX Upgrade

Plasma disruptions pose an intolerable risk to large tokamaks, such as ITER. If a disruption can no longer be avoided, ITER’s last line of defense will be the Shattered Pellet Injection. An experimental test bench was created at ASDEX Upgrade to inform the design decisions for controlling the shattering of the pellets and develop the techniques for the generation of the fragment distributions necessary for optimal disruption mitigation. In an effort to analyze the videos resulting from the more than 1000 tests and determine the impact of different settings on the resulting shard cloud, an analysis pipeline, based on traditional computer vision (CV), was created. This pipeline enabled the analysis of 173 of the videos, but at the same time showed the limits of traditional CV when applied in applications with a highly heterogeneous dataset such as this. We created a machine learning-based (ML) alternative as a drop-in replacement to the original image processing code using a semantic segmentation model to exploit the innate adaptability and robustness of deep learning models. This model is capable of labeling the entire dataset quickly, accurately and reliably. This contribution details the implementation of the ML model and the current state and future plans of the project.

Dynamic Profiling and Prediction of Antibody Response to SARS-CoV-2 Booster-Inactivated Vaccines by Microsample-Driven Biosensor and Machine Learning.

Knowledge of the antibody response to the third dose of inactivated SARS-CoV-2 vaccines is crucial because it is the subject of one of the largest global vaccination programs. This study integrated microsampling with optical biosensors to profile neutralizing antibodies (NAbs) in fifteen vaccinated healthy donors, followed by the application of machine learning to predict antibody response at given timepoints. Over a nine-month duration, microsampling and venipuncture were conducted at seven individual timepoints. A refined iteration of a fiber optic biolayer interferometry (FO-BLI) biosensor was designed, enabling rapid multiplexed biosensing of the NAbs of both wild-type and Omicron SARS-CoV-2 variants in minutes. Findings revealed a strong correlation (Pearson r of 0.919, specificity of 100%) between wild-type variant NAb levels in microsamples and sera. Following the third dose, sera NAb levels of the wild-type variant increased 2.9-fold after seven days and 3.3-fold within a month, subsequently waning and becoming undetectable after three months. Considerable but incomplete evasion of the latest Omicron subvariants from booster vaccine-elicited NAbs was confirmed, although a higher number of binding antibodies (BAbs) was identified by another rapid FO-BLI biosensor in minutes. Significantly, FO-BLI highly correlated with a pseudovirus neutralization assay in identifying neutralizing capacities (Pearson r of 0.983). Additionally, machine learning demonstrated exceptional accuracy in predicting antibody levels, with an error level of <5% for both NAbs and BAbs across multiple timepoints. Microsample-driven biosensing enables individuals to access their results within hours of self-collection, while precise models could guide personalized vaccination strategies. The technology's innate adaptability means it has the potential for effective translation in disease prevention and vaccine development.

Agile robotic inspection of steel structures: A bicycle‐like approach with multisensor integration

AbstractThis paper introduces an innovative and streamlined design of a robot, resembling a bicycle, created to effectively inspect a wide range of ferromagnetic structures, even those with intricate shapes. The key highlight of this robot lies in its mechanical simplicity coupled with remarkable agility. The locomotion strategy hinges on the arrangement of two magnetic wheels in a configuration akin to a bicycle, augmented by two independent steering actuators. This configuration grants the robot the exceptional ability to move in multiple directions. Moreover, the robot employs a reciprocating mechanism that allows it to alter its shape, thereby surmounting obstacles effortlessly. An inherent trait of the robot is its innate adaptability to uneven and intricate surfaces on steel structures, facilitated by a dynamic joint. To underscore its practicality, the robot's application is demonstrated through the utilization of an ultrasonic sensor for gauging steel thickness, coupled with a pragmatic deployment mechanism. By integrating a defect detection model based on deep learning, the robot showcases its proficiency in automatically identifying and pinpointing areas of rust on steel surfaces. The paper undertakes a thorough analysis, encompassing robot kinematics, adhesive force, potential sliding and turn‐over scenarios, and motor power requirements. These analyses collectively validate the stability and robustness of the proposed design. Notably, the theoretical calculations established in this study serve as a valuable blueprint for developing future robots tailored for climbing steel structures. To enhance its inspection capabilities, the robot is equipped with a camera that employs deep learning algorithms to detect rust visually. The paper substantiates its claims with empirical evidence, sharing results from extensive experiments and real‐world deployments on diverse steel bridges, situated in both Nevada and Georgia. These tests comprehensively affirm the robot's proficiency in adhering to surfaces, navigating challenging terrains, and executing thorough inspections. A comprehensive visual representation of the robot's trials and field deployments is presented in videos accessible at the following links: https://youtu.be/Qdh1oz_oxiQ and https://youtu.be/vFFq79O49dM.

Measles Virus-Induced Host Immunity and Mechanisms of Viral Evasion

The immune system deploys a complex network of cells and signaling pathways to protect host integrity against exogenous threats, including measles virus (MeV). However, throughout its evolutionary path, MeV developed various mechanisms to disrupt and evade immune responses. Despite an available vaccine, MeV remains an important re-emerging pathogen with a continuous increase in prevalence worldwide during the last decade. Considerable knowledge has been accumulated regarding MeV interactions with the innate immune system through two antagonistic aspects: recognition of the virus by cellular sensors and viral ability to inhibit the induction of the interferon cascade. Indeed, while the host could use several innate adaptors to sense MeV infection, the virus is adapted to unsettle defenses by obstructing host cell signaling pathways. Recent works have highlighted a novel aspect of innate immune response directed against MeV unexpectedly involving DNA-related sensing through activation of the cGAS/STING axis, even in the absence of any viral DNA intermediate. In addition, while MeV infection most often causes a mild disease and triggers a lifelong immunity, its tropism for invariant T-cells and memory T and B-cells provokes the elimination of one primary shield and the pre-existing immunity against previously encountered pathogens, known as "immune amnesia".

Microbiota-Driven Activation of Intrahepatic B Cells Aggravates NASH Through Innate and Adaptive Signaling.

Background and AimsNonalcoholic steatohepatitis is rapidly becoming the leading cause of liver failure and indication for liver transplantation. Hepatic inflammation is a key feature of NASH but the immune pathways involved in this process are poorly understood. B lymphocytes are cells of the adaptive immune system that are critical regulators of immune responses. However, the role of B cells in the pathogenesis of NASH and the potential mechanisms leading to their activation in the liver are unclear.Approach and ResultsIn this study, we report that NASH livers accumulate B cells with elevated pro‐inflammatory cytokine secretion and antigen‐presentation ability. Single‐cell and bulk RNA sequencing of intrahepatic B cells from mice with NASH unveiled a transcriptional landscape that reflects their pro‐inflammatory function. Accordingly, B‐cell deficiency ameliorated NASH progression, and adoptively transferring B cells from NASH livers recapitulates the disease. Mechanistically, B‐cell activation during NASH involves signaling through the innate adaptor myeloid differentiation primary response protein 88 (MyD88) as B cell–specific deletion of MyD88 reduced hepatic T cell–mediated inflammation and fibrosis, but not steatosis. In addition, activation of intrahepatic B cells implicates B cell–receptor signaling, delineating a synergy between innate and adaptive mechanisms of antigen recognition. Furthermore, fecal microbiota transplantation of human NAFLD gut microbiotas into recipient mice promoted the progression of NASH by increasing the accumulation and activation of intrahepatic B cells, suggesting that gut microbial factors drive the pathogenic function of B cells during NASH.ConclusionOur findings reveal that a gut microbiota–driven activation of intrahepatic B cells leads to hepatic inflammation and fibrosis during the progression of NASH through innate and adaptive immune mechanisms.

Abstract 1102: Integrated approach towards defining mechanism based combinations to guide clinical development of glutaminase inhibitors

Abstract Metabolic dysregulation is a hallmark of cancer, and recently, increasing evidence has shown a critical role for glutamine metabolism to support the bioenergetics and biosysnthetic needs of tumor and immune cells. We have previously disclosed the development of the GLS1 inhibitor, IPN60090, which is currently advancing through Phase 1 clinical studies (NCT03894540). Metabolic plasticity has been suggested to confer adaptive resistance to metabolic inhibitors and defines mechanisms that could be exploited to enhance therapeutic benefit. To address the clinical problems of innate drug resistance and adaptation, we interrogated metabolic and adaptive responses to IPN60090 in vitro and in vivo. We and others have previously identified KEAP1/NFE2L2 mutant non-small cell lung cancer models as sensitive to GLS1 inhibition. Through an integrated approach, including metabolic, transcriptomic, and proteomic analysis, we have identified molecular pathways that confer resistance to GLS1 inhibition. Nodes in these pathways that drive aquired resistance, may serve as additional patient stratification biomarkers in subsets on NSCLC or provide opportunites for further drug development. Additionally, unbiased in vivo functional genomics screening identified mulitple signaling pathways that act as critical nodes governing resistance to GLS1 inhibition. Drug combinations were tested in vitro and in vivo to identify those that are most synergistic with IPN60090. We found that PI3K/AKT/mTOR signaling is a major contributor to IPN60090 resistance, and we demonstrate that dual targeting of GLS1 and PI3K/AKT/mTOR signaling in tumors with KEAP1/NFE2L2 mutations results in synergistic anti-tumor efficacy. IPN60090, dosed in combination with inhibitors of these pathways yields regressions and off-treatment, durable responses in preclinical models of KEAP1-mutant NSCLC. Based on these data, combination strategies are being developed for Phase 1b expansion cohorts. Citation Format: Jeffrey J. Kovacs, Nakia D. Spencer, Christopher A. Bristow, Alessandro Carugo, Virginia Giluiani, Meredith A. Miller, Angela Harris, Ningping Feng, Michael L. Soth, Kang Le, Elisa de Stanchina, Charles M. Rudin, Giulio Draetta, Timothy A. Yap, Philip Jones, Timothy Heffernan. Integrated approach towards defining mechanism based combinations to guide clinical development of glutaminase inhibitors [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1102.

Heart rate variability in Konik and purebred Arabian horses in response to different predator vocalisations

The current predation threat of domestic horses is generally low, and horses do not know predators' frightening cues. We studied whether horses still recognise predation threats. The aim of the study was to analyse the emotional response of purebred Arabian horses (Arabian) and Polish Konik horses (Konik) to an Arabian panther (Panthera pardus nimr) (panther) growl and a grey wolf (Canis lupus) (wolf) howl. Panther vocalisation was known to Arabian ancestors, whereas ancestors of Konik knew wolf vocalisation. The response to the howls of golden jackals (Canis aureus) (jackal), which did not prey on equids, was also studied comparatively. Two groups of 10 adult horses of each breed were subject to predator sounds of one predator daily for 5 min during a turn out on pasture. The test was performed for 18 days in total. The sound of each predator was interchangeably featured from one loudspeaker for 3 days followed by four loudspeakers simultaneously to imitate a group of predators for 3 days. The horses' emotional agitation in response to the sounds was measured based on the parameters of heart rate variability (HRV) using telemetric devices. The results showed that the predators' sounds were identified by horses as stressful or neutral. Horses generally retained their anti-predator responses even in the current habitat, which typically lacks predation cues. The results are not always coherent and may demonstrate that the response is somewhat attenuated. The wolf howl elicited a stronger response in Koniks. The panther growl more strongly influenced Arabians, whereas the jackal howl minimally elicited an agitation in the horses. The differentiated response of the two horse breeds to the three predator species suggests that the response is an innate adaptation to the predation risk in the habitat of the breed ancestors. This response occurs regardless of the emotional arousal specific to a breed, and the frightening cue is not the sound per se but the possible attack of predators. Horses display a type of understanding of the sound meaning. Their HRV response seems to be adequate for the threat signalised by the sound.

Deciphering the Cold Adaptive Mechanisms in Pseudomonas psychrophila MTCC12324 Isolated from the Arctic at 79° N.

Psychrophiles, host of cold environments, have been successfully undergoing the process of evolution by which they have acquired innate adaptations to withstand the unfavorable effects of low temperature. Psychrophiles renders immense opportunity to explore the underlying mechanisms of cold adaptation. The present study focused to explore the cold adaptive mechanisms of Pseudomonas psychrophila MTCC12324, a facultative psychrophilic bacterium isolated from the Ny-Alesund, an island in the Svalbard Archipelago (79°55' N, 11°56' E) in the Arctic. Whole genome sequencing of P. psychrophila MTCC12324 and its analysis revealed the redundant nature of genome and identified several cold acclimation genes including cold shock proteins, and chaperones involved in the adaptive mechanism to thrive in the cold environment. Comparative proteome analysis of P. psychrophila MTCC12324 at 4°C and 25°C has thrown lights on the metabolic pathways and cellular processes adopted to withstand the cold environment. Basic survival pathways and factors involved in energy metabolism were found to be unaltered whereas stress response factors, enzymes involved in fatty acid elongation and cold-adapted chaperones were found to be enhanced towards cold stress. The present study facilitates recognition of crucial factors including polyunsaturated fatty acid biosynthesis, mRNA chaperones, and other cold-inducible proteins which favors the bacteria in conferring cold adaptation.

TBK1, a central kinase in innate immune sensing of nucleic acids and beyond.

Sensing of intracellular and extracellular environments is one of the fundamental processes of cell. Surveillance of aberrant nucleic acids, derived either from invading pathogens or damaged organelle, is conducted by pattern recognition receptors (PRRs) including RIG-I-like receptors, cyclic GMP-AMP synthase, absent in melanoma 2, and a few members of toll-like receptors. TANK-binding kinase 1 (TBK1), along with its close analogue I-kappa-B kinase epsilon, is a central kinase in innate adaptor complexes linking activation of PRRs to mobilization of transcriptional factors that transcribe proinflammatory cytokines, type I interferon (IFN-α/β), and myriads interferon stimulated genes. However, it still remains elusive for the precise mechanisms of activation and execution of TBK1 in signaling platforms formed by innate adaptors mitochondrial antiviral signaling protein (MAVS), stimulator of interferon genes protein (STING), and TIR-domain-containing adapter-inducing interferon-β (TRIF), as well as its complex regulations. An atlas of TBK1 substrates is in constant expanding, setting TBK1 as a key node of signaling network and a dominant player in contexts of cell biology, animal models, and human diseases. Here, we review recent advancements of activation, regulations, and functions of TBK1 under these physiological and pathological contexts.

Must all signals be evolved? A proposal for a new classification of communicative acts.

While signals in evolutionary biology are usually defined as "acts or traits that have evolved because of their effect on others", work on gestures and vocalizations in various animal taxa have revealed population- or even individual-specific meanings of social signals. These results strongly suggest that communicative acts that are like signals with regard to both form and function (meaning) can also be acquired ontogenetically, and we discuss direct evidence for such plasticity in captive settings with rich opportunities for repeated social interactions with the same individuals. Therefore, in addition to evolved signals, we can recognize invented signals that are acquired during ontogeny (either through ontogenetic ritualization or social transmission). Thus, both gestures and vocalizations can be inventions or innate adaptations. We therefore propose to introduce innate versus invented signals as major distinct categories, with invented signals subdivided into dyad-specific and cultural signals. We suggest that elements of some signals may have mixed origins, and propose criteria to recognize acquired features of signals. We also suggest that invented signals may be most common in species with intentional communication, consistent with their ubiquity in humans, and that the ability to produce them was a necessary condition for the evolution of language. This article is categorized under: Cognitive Biology > Evolutionary Roots of Cognition Linguistics > Evolution of Language Psychology > Comparative Psychology.

MyD88 mediates the pathogenesis of dermatitis in SHARPIN-deficient mice by regulating TNF production on the skin

Abstract Mice deficient in SHANK-associated RH domain–interacting protein (SHARPIN), a component of the linear ubiquitin chain assembly complex (LUBAC), develop a spontaneous inflammatory disorder with pathologic hallmarks similar to atopic dermatitis and psoriasis in humans. Previous studies identified the crucial role of components of the TNF and IL-1 signaling pathways in the progression of disease in SHARPIN-deficient mice. However, an innate adaptor or sensor that relates to the disease progression has remained unknown. In this study, we found that the genetic ablation of myeloid differentiation primary response 88 (MyD88) completely rescued skin inflammation in SHARPIN-deficient (Sharpincpdm) mice. Systemic inflammation and immune cell dysregulation were also rescued. Fibroblasts derived from SharpincpdmMyd88−⁄− micefailed to provide protection against TNF-induced cell death. SharpincpdmMyd88−⁄− mice had reduced TNF production in their skin. Furthermore, depletion of the microbiota through the oral administration of antibiotics partially rescued both the skin inflammation and systemic inflammation, demonstrating a role for the gut microbiota in SHARPIN-deficient mice. Our findings suggest a detrimental role for the innate immune adaptor MyD88 in instigating skin inflammation in Sharpincpdm mice.

Abstract B66: Targeted activation of innate immune adaptors MAVS and STING promotes antitumor responses in colorectal cancer models

Abstract While the use of immune checkpoint blockade (ICB) has gained significant traction as a viable therapy for certain cancers, many more cancers remain refractory to immunotherapies due to their highly immunosuppressive microenvironment. This is perhaps best exemplified in the largest group of cancers, colorectal carcinomas (CRCs) where ~90% of CRCs (non-microsatellite instable) are unresponsive to ICBs. Recent studies have indicated that this lack of response may be due to mutations and loss of genes that mediate the innate immune responses, such as MAVS (Mitochondrial Antiviral Signaling Protein) and STING (Stimulator of Interferon Genes). We hypothesized that delivery of active versions of these genes would elicit strong interferon signaling that could reverse the tumor immunosuppressive environment, to effectively create immune “hotspots” and allow for successful immune-mediated tumor regression. As a proof-of-concept, we developed doxycycline-inducible activated forms STING and MAVS in mouse CRC models (MC38 and CT26 lines), and found the expression of MAVS or STING resulted in robust activation of interferon-stimulated genes and other cytokines/chemokines. To determine if delivery of these genes could be achieved in vivo, we developed cell lines capable of resisting the anti-viral effect of interferon stimulation. This system allowed us to generate adenoviral vectors that express active forms of MAVS and more recently, STING. We found that Ad-MAVS strongly elicited interferon signaling in a variety of cell types and tumor lines, suggesting its effectiveness as an interferon stimulatory agent. To determine its anti-tumor effect, we intra-lesionally injected tumors and observed strongly inhibited tumor growth and enhanced T-cell infiltration in established tumor masses in vivo. This correlated with enhanced anti-tumor specific T cell responses, but not humoral responses. The induction of local and systemic immunity was confirmed in a bi-lateral treatment model that demonstrated the impact of systemic immunity by MAVS induction. Furthermore, we found that MAVS-induced tumor regression was lost in NOD/SCID mice, suggesting that the induction of tumor-specific T-cell adaptive immunity was critical to anti-tumor responses. Lastly, we found the activation of MAVS resulted in upregulated expression of PD-L1 on tumor cells, suggesting combinatorial treatments of checkpoint blockades therapy with targeted MAVS activation can result in synergistic effects. In conclusion, our data demonstrated the importance of innate immunity activation by MAVS or STING pathway in generating immune hotspots in colorectal tumor microenvironment, which promotes antitumor adaptive T cell responses and tumor growth regression. Citation Format: Li-Chung Tsao, Herbert Kim Lyerly, Zachary Hartman, Gabriel Deleon. Targeted activation of innate immune adaptors MAVS and STING promotes antitumor responses in colorectal cancer models [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2017 Oct 1-4; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2018;6(9 Suppl):Abstract nr B66.

Abstract 124: The soluble factor SPINK1 is a biomarker of therapeutically damaged TME that confers acquired resistance and an exploitable molecular target in human cancer medicine

Abstract Clinical efficacy of most anticancer treatments is frequently compromised by the development of cancer resistance arising as either a consequence of innate adaptation of cancer cells, or acquired fitness conferred by extracellular factors generated by the surrounding tumor microenvironment (TME). Of note, stromal cells in the TME become overtly senescent upon genotoxic treatment and develop a senescence-associated secretory phenotype (SASP), the latter characterized by secretion of numerous soluble factors that are responsible for disease exacerbation. We recently disclosed significant upregulation of the serine protease inhibitor Kazal type I (SPINK1) in human prostate stroma after chemotherapy of prostate cancer (PCa) patients. SPINK1, originally reported as a pancreatic secretory trypsin inhibitor, prevents pancreatic tissues from protease degradation with expression normally restricted to gastrointestinal (GI) tract organs. However, the influence of SPINK1 as a TME-derived soluble factor on cancer progression remains unexplored. In this study, we defined the expression modality and biological roles of SPINK1 in a therapy-damaged and functionally activated TME under therapeutic pressure. Distinct from a recent finding that revealed the involvement of a regulatory circuit comprising HNF4G and HNF1A in the PCa-GI transcriptome, we found that SPINK1 expression in human stromal cells is regulated by alternative transcriptional factors including NF-κB and C/EBP in response to chemotherapy. TME-released SPINK1 can significantly enhance the aggressiveness of PCa cells including accelerated proliferation, increased migration, elevated invasion and more importantly, enhanced chemoresistance. SPINK1 triggers a typical epithelial-mesenchymal transition in cancer cells, a process mediated by EGFR/PI3K/Akt and MAPK/Erk signaling. Further, SPINK1 remarkably reprograms PCa cells by inducing a cancer stem cell (CSC)-like phenotype and promoting transdifferentiation of cancer cells toward neovasculature. As supporting evidence, our preclinical studies suggested that SPINK1 expression in the prostate TME substantially promoted cancer survival and disease progression. Clinical investigation revealed increased expression of SPINK1 in the stroma of multiple organ types including the prostate, lung and breast of cancer patients after chemotherapy, implying the systemic induction of SPINK1 by anticancer agents. Thus, our study demonstrates that SPINK1 is a soluble biomarker of therapeutically damaged TME and represents a potentially exploitable molecular target in human cancer clinics. Citation Format: Yu Sun. The soluble factor SPINK1 is a biomarker of therapeutically damaged TME that confers acquired resistance and an exploitable molecular target in human cancer medicine [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 124.

Abstract 487: Myocardial Infarction is Associated with Changes in Innate Immunity that Drive Breast Cancer Progression

Cardiovascular (CV) disease and cancer-specific mortality compete as the leading causes of mortality in women with early-stage breast cancer. Following treatment for early-stage disease, breast cancer patients are 5 times more likely to develop CV complications such as myocardial infarction (MI) compared to age-matched controls, due to the ‘cardiotoxic’ effects of anti-cancer therapies on CV tissues (e.g. chemotherapy, radiation). As CV disease and cancer progression share many common risk factors and etiologies, we hypothesized that CV disease may also influence cancer progression. To investigate this potential cross-disease communication, we developed a mouse model using the left anterior descending (LAD) coronary artery permanent ligation technique of MI in the E0771 orthotopic breast cancer model. LAD ligation significantly increased tumor growth over 20 days compared to sham surgery controls (SHAM) (p&lt;0.001). This was associated with an altered tumor microenvironment and tumor immune cell landscape, most notably increased proportions of CD11b+Ly6C+Ly6G- myeloid cells in LAD vs SHAM (p&lt;0.01). RNA and ATAC sequencing of these cells from the bone marrow, circulation, and tumor showed distinct functional and chromatin-based changes following LAD ligation, including increased expression of genes associated with cell proliferation, mitochondria and lipid transport, and enrichment of transcription factor binding motifs within differentially accessible regions, including PU.1, Ets1, Runx1 and Cebp. Using monocyte adoptive transfer experiments, we also show that recruitment of bone marrow monocytes to the tumor microenvironment was increased in LAD vs SHAM mice. Collectively, these findings suggest that MI can accelerate breast cancer progression and provide insight into molecular mechanisms driving innate immune adaptations that promote cross-disease communication.

Reshaping of the Dendritic Cell Chromatin Landscape and Interferon Pathways during HIV Infection

Myeloid dendritic cells (DCs) have the innate capacity to sense pathogens and orchestrate immune responses. However, DCs do not mount efficient immune responses to HIV-1, primarily due to restriction of virus reverse transcription, which prevents accumulation of viral cDNA and limits its detection through the cGAS-STING pathway. By allowing reverse transcription to proceed, we find that DCs detect HIV-1 in distinct phases, before and after virus integration.Blocking integration suppresses, but does notabolish, activation of the transcription factor IRF3, downstream interferon (IFN) responses, and DC maturation. Consistent with two stages of detection, HIV-1 "primes" chromatin accessibility of innate immune genes before and after integration. Once primed, robust IFN responses can be unmasked byagonists of the innate adaptor protein, MyD88, through a process that requires cGAS, STING, IRF3, and nuclear factor κB. Thus, HIV-1 replication increases material available for sensing, and discrete inflammatory inputs tune cGAS signaling to drive DC maturation.

Complex of reactions providing protective behavior of leaf beetles (Coleoptera, Chrysomelidae)

Protective behavior of 25 background species of leaf beetles (Coleoptera, Chrysomelidae) were studied in the conditions of the Samara Region during 1974-2014. Strategy of protective behavior is focused on conservation, enhancement and resettlement of a type in space. It includes 2 blocks of different and difficult reactions. There is passive protection, presented by 25 types of reactions and active protection, presented by more than 45 types of motive implications. Passive reactions do not demand an additional expenditure of efforts and energy. They are presented, mainly, by immobile posture of masking, concealment and others. On the contrary, active protection isnt possible without expense of additional efforts. In addition, active protection is more difficult, and also includes a series from several protective reactions, it is more effective. It is related to behavior of imago and larvas protection. Protective behavior can be individual and group. Protective behavior is closely bound to other functional behavioral blocks. There is trophism, communication and reproduction. Leaf beetles have a system of innate morphological, anatomic and physiological adaptations allowing them to experience many negative impacts of the environment.

Abstract B04: Intralesional vaccination with Ad-MAVS alters the immunosuppressive tumor microenvironment and elicits robust anti-tumor immunity in non-immunogenic cancers

Abstract Background and Hypothesis: While the use of immune checkpoint blockade has gained significant traction as a viable therapy for certain cancers, many tumors remain refractory to these immunotherapies due to their highly immunosuppressive microenvironment. As a novel approach to boost local anti-tumor immunity and sensitize cancers to these therapies, we constructed an adenoviral (Ad) vector to overexpress the innate signaling adaptor molecule, Mitochondrial Antiviral Signaling protein (MAVS). Overexpression of MAVS results in innate immune signaling downstream of the RIG-I like receptors (RLRs) resulting in the activation of multiple signaling pathways as well as the release of a Th1 cytokine milieu and type I interferons. We therefore hypothesized that adenoviral delivery of MAVS into the tumor microenvironment would produce robust inflammatory signaling which would alter the tumor microenvironment to both efficiently suppress tumor growth and stimulate anti-tumor immune responses. Methods: In order to determine the possible effect MAVS would have on different cell types within the tumor microenvironment we infected a range of primary and cancer cells types (Bone-marrow derived dendritic cells (BMDCs), primary fibroblasts, as well as 5 different murine tumor cell lines representing breast-4T1, colon-CT26CL25, bladder-MB49, melanoma-B16F10, and prostate-TRAMPC2) with Ad-MAVS or a control Ad. Innate immune responses were measured via luminex ELISA, microarray, and RT-qPCR. The impact on tumor growth was assessed by MTT and expression of PD-L1 by FACS. Lastly, we measured the anti-tumor effect and adaptive immune responses of animals treated with various combinations of Ad-MAVS, a control Ad, and PD-L1 targeting antibodies in vivo utilizing various immunocompetent and immune-compromised mouse models. Results: In vitro infection of BMDCs, MEFs, and murine tumor cell lines demonstrated that MAVS expression elicited widespread production of cytokines and chemokines indicative of a type I immune response (such as IL-12, TNF-alpha, RANTES, etc.) and interferon beta. These profound responses also triggered immune regulatory feedback expression of PD-L1, enabling their targeting by PD-L1 antibodies. Additionally, we found that Ad-MAVS infection of tumor cell lines significantly inhibited their proliferation in vitro, likely via interferon pathways. In vivo, treatment with Ad-MAVS also elicited robust stimulation of innate immune responses and numerous type I inflammatory cytokines. Moreover, in multiple types of tumor bearing animals, we found that a single intralesional treatment of Ad-MAVS significantly prolonged overall survival and was capable of eliciting systemic anti-tumor adaptive immunity to non-immunogenic tumors. Conclusion: Our study identifies that MAVS expression can elicit the production of a significant number of Th1-type cytokines, chemokines, and interferon-beta in multiple cell types within the tumor microenvironment. We also demonstrate that overexpression of MAVS results in the direct suppression of tumor cell growth and stimulation of tumor PD-L1 expression. These activities were also observed in vivo, as intralesional injection of Ad-MAVS elicited Th1 cytokines and anti-tumor immunity in multiple tumor types, which correlated with enhanced systemic T-cell responses to tumor-specific epitopes. Collectively, these data demonstrate that intralesion injection of Ad-MAVS alters the tumor microenvironment to stimulate anti-tumor immunity and potentially sensitizes non-immunogenic tumors to PD-L1 therapies. As such, Ad-MAVS represents a novel approach of stimulating anti-tumor immunity via activation of innate adaptor molecules to improve immunogenic anti-tumor responses and potentially the therapeutic efficacy of PD-L1 immune checkpoint blockade. Citation Format: Gabriel De Leon, Tao Wang, Pankaj Agarwal, H. Kim Lyerly, Zachary Hartman. Intralesional vaccination with Ad-MAVS alters the immunosuppressive tumor microenvironment and elicits robust anti-tumor immunity in non-immunogenic cancers. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology and Immunotherapy; 2016 Oct 20-23; Boston, MA. Philadelphia (PA): AACR; Cancer Immunol Res 2017;5(3 Suppl):Abstract nr B04.

Molecular Mechanisms That Underlie the Dynamic Adaptation of Innate Monocyte Memory to Varying Stimulant Strength of TLR Ligands.

In adaptation to rising stimulant strength, innate monocytes can be dynamically programed to preferentially express either pro- or anti-inflammatory mediators. Such dynamic innate adaptation or programing may bear profound relevance in host health and disease. However, molecular mechanisms that govern innate adaptation to varying strength of stimulants are not well understood. Using lipopolysaccharide (LPS), the model stimulant of toll-like-receptor 4 (TLR4), we reported that the expressions of pro-inflammatory mediators are preferentially sustained in monocytes adapted by lower doses of LPS, and suppressed/tolerized in monocytes adapted by higher doses of LPS. Mechanistically, monocytes adapted by super-low dose LPS exhibited higher levels of transcription factor, interferon regulatory factor 5 (IRF5), and reduced levels of transcriptional modulator B lymphocyte-induced maturation protein-1 (Blimp-1). Intriguingly, the inflammatory monocyte adaptation by super-low dose LPS is dependent upon TRAM/TRIF but not MyD88. Similar to LPS, we also observed biphasic inflammatory adaptation and tolerance in monocytes challenged with varying dosages of TLR7 agonist. In sharp contrast, rising doses of TLR3 agonist preferentially caused inflammatory adaptation without inducing tolerance. At the molecular level, the differential regulation of IRF5 and Blimp-1 coincides with unique monocyte adaptation dynamics by TLR4/7 and TLR3 agonists. Our study provides novel clue toward the understanding of monocyte adaptation and memory toward distinct TLR ligands.

Differential Effects on Intestinal Adaptation Following Exogenous Glucagon-Like Peptide 2 Therapy With and Without Enteral Nutrition in Neonatal Short Bowel Syndrome [Formula: see text

We aim to study the efficacy of exogenously administered glucagon-like peptide 2 (GLP-2) on intestinal adaptation in 2 preclinical models of neonatal short bowel syndrome (SBS) according to remnant intestinal anatomy, with and without ileum. Furthermore, we aim to determine if this adaptive effect was potentiated with enteral nutrition (EN). Neonatal piglets were block-randomized to 75% mid-intestinal (JI group, retains ileum) or distal-intestinal (JC group, has no ileum) resection or no resection (sham control) and GLP-2 treatment (11 nmol/kg/d) or saline control for 7 days. Piglets received nutrition support, either 100% parenteral nutrition (PN; 0% EN, n = 32 in total) or 80% PN + 40% EN (n = 28 in total). Adaptation was assessed by morphological and histological changes, as well as RT quantitative polymerase chain reaction of nutrient transporters and tight junctional proteins and fat absorption. Data are analyzed by 3-way analysis of variance (ANOVA) and 2-way ANOVA per EN level. GLP-2 treatment lengthened villi, deepened crypts, and improved intestinal weight in the remnant intestine of JC piglets. EN was a more potent adaptive stimulus for JI piglets. Small intestinal lengthening occurred only in the JI group, when given EN. There was no difference in total fat absorption and messenger RNA expression of nutrient transporters and tight junctional proteins. GLP-2 administration augmented structural adaptation in JC piglets with distal intestinal resection. Given JI anatomy, further stimulation by GLP-2 treatment over innate adaptation and stimulation by EN was modest and restricted to ileum. The differential effect of GLP-2 in neonatal SBS, depending on remnant anatomy, has important implications for clinical translation and planning of clinical trials.

Liver fibrosis occurs through dysregulation of MyD88‐dependent innate B‐cell activity

Chronic liver disease mediated by activation of hepatic stellate cells (HSCs) leads to liver fibrosis. Here, we postulated that the immune regulatory properties of HSCs might promote the profibrogenic activity of B cells. Fibrosis is completely attenuated in carbon tetrachloride-treated, B cell-deficient µMT mice, showing that B cells are required. The retinoic acid produced by HSCs augmented B-cell survival, plasma cell marker CD138 expression, and immunoglobulin G production. These activities were reversed following addition of the retinoic acid inhibitor LE540. Transcriptional profiling of fibrotic liver B cells revealed increased expression of genes related to activation of nuclear factor κ light chain enhancer of activated B cells, proinflammatory cytokine production, and CD40 signaling, suggesting that these B cells are activated and may be acting as inflammatory cells. Biological validation experiments also revealed increased activation (CD44 and CD86 expression), constitutive immunoglobulin G production, and secretion of the proinflammatory cytokines tumor necrosis factor-α, monocyte chemoattractant protein-1, and macrophage inflammatory protein-1α. Likewise, targeted deletion of B-cell-intrinsic myeloid differentiation primary response gene 88 signaling, an innate adaptor with involvement in retinoic acid signaling, resulted in reduced infiltration of migratory CD11c(+) dendritic cells and Ly6C(++) monocytes and, hence, reduced liver pathology. Liver fibrosis occurs through a mechanism of HSC-mediated augmentation of innate B-cell activity. These findings highlight B cells as important "first responders" of the intrahepatic immune environment.