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Nivolumab and pembrolizumab, anti–programmed cell death protein 1 monoclonal antibodies, have revolutionized oncology but are expensive. Using an interventional pharmacoeconomic approach, these drugs can be administered less often to reduce costs and increase patient convenience while maintaining efficacy. Both drugs are good candidates for less frequent dosing because of long half-lives and no evidence of a relationship of dose to efficacy. Established population pharmacokinetic models for both nivolumab and pembrolizumab were used to simulate profiles for multiple dosing regimens on 1000 randomly generated virtual patients. Simulations were initially performed on standard dose regimens to validate these in silico predictions. Next, simulations of nivolumab 0.3 mg/kg every 3 weeks revealed that >95% of patients maintained ≥1.5 μg/mL at steady state, which was inferred as the minimum effective concentration (MEC) for both drugs. Various alternative dosing regimens were simulated for both drugs to determine which regimen(s) can maintain this MEC in >95% of patients. Extended dosing regimens of nivolumab 240 mg every 4 weeks and 480 mg every 8 weeks along with pembrolizumab 200 mg every 6 weeks were simulated, showing that >95% of patients maintained MEC or greater. These simulations demonstrate the potential to reduce drug exposure by at least 50%, thus substantially reducing patient visits (as well as costs), while maintaining equivalent efficacy. These models provide the scientific justification for an ongoing prospective randomized clinical trial comparing standard interval fixed dosing with extended interval fixed dosing, and ultimately an efficacy-driven comparative trial.

Tissue damage occurs in COVID-19 patients due to nsp3-induced Fas-FasL interaction/TNF-related apoptosis. Presently, possible therapeutic-drug, nigellidine against was screened by bioinformatics studies COVID-19. Atomic-Contact-Energy (ACE) and binding-blocking effects were explored of nigellidine (Nigella sativa L.) in the active/catalytic sites of viral-protein nsp3 and host inflammatory/apoptotic signaling-molecules Fas/TNF receptors TNFR1/TNFR2. A control binding/inhibition of Oseltamivir to influenza-virus neuraminidase was compared here. In AutoDock, Oseltamivir binding-energy (BE) and inhibition-constant (KI) was −4.12 kcal/mol and 959.02. The ACE values (PatchDock) were −167.02/-127.61/-124.91/-122.17/-54.81/-47.07. The nigellidine BE/KI with nsp3 was −7.61 and 2.66, respectively (ACE values were −221.40/-215.62/-113.28). Nigellidine blocked FAS dimer by binding with a BE value of −7.41 kcal/mol. Its strong affinities to TNFR1 (-6.81) and TNFR2 (-5.1) are demonstrated. Our present data suggest that nigellidine may significantly block the TNF-induced inflammatory/Fas-induced apoptotic death-signaling in comparison with a positive-control drug Oseltamivir. Further studies are necessary before proposing nigellidine as medical drug.

The COVID-19 pandemic is a highly dramatic concern for mankind. In Italy, the pandemic exerted its major impact throughout the period of February to June 2020. To date, the awkward amount of more than 134,000 deaths has been reported. Yet, post-mortem autopsy was performed on a very modest number of patients who died from COVID-19 infection, leading to a first confirmation of an immune-thrombosis of the lungs as the major COVID-19 pathogenesis, likewise for SARS. Since then (June-August 2020), no targeted early therapy considering this pathogenetic issue was approached. The patients treated with early anti-inflammatory, anti-platelet, anticoagulant and antibiotic therapy confirmed that COVID-19 was an endothelial inflammation with immuno-thrombosis. Patients not treated or scarcely treated with the most proper and appropriate therapy and in the earliest, increased the hospitalization rate in the intensive care units and also mortality, due to immune-thrombosis from the pulmonary capillary district and alveoli. The disease causes widespread endothelial inflammation, which can induce damage to various organs and systems. Therapy must be targeted in this consideration, and in this review, we demonstrate how early anti-inflammatory therapy may treat endothelia inflammation and immune-thrombosis caused by COVID-19, by using drugs we are going to recommend in this paper.

Ayurveda Immune boosters (AIB) are Rasayana drugs which increase the number of white blood cells and train them to fight against microbes causing diseases Some of them kill microorganisms, increase immunoglobulin, repair of DNA of an inflamed cell, increase Agni, and conditioning the gut to increase the beneficial gut microbiota This review aimed to generate scientific evidence for Ayurveda Immune boosters (AIB) are effective to prevent and cure COVID-19 infection or it is a myth through Ayurveda and modern literature review The properties of Rasayan are reviewed from Ayurveda Literature The immunopharmacology, mechanism of immuno-booster and Rasayana herbs are reviewed from current medical literature for its subclinical and clinical evidence Ayush kwatha and single herb like Guduchi (500-1000mg) extract, Aswagandha powder (3-5gm), and Haridra milk/gargling recommended as AIB by the Ministry AYUSH are safe and effective for prevention and cure of COVID-19 with sufficient preclinical and some Randomised clinical trials (RCT) evidence So, the myth of Traditional Ayurveda herbs won’t boost our immunity to the virus is not true rather Ayurveda Medication (AIB) can boost our immunity to fight against COVID-19 More RCT evidence is required for clinical practice © 2021, Radiance Research Academy All rights reserved

The aim of the article is to generalize information on possible molecular targets that ensure the implementation of the immunotoxic effects of drugs for the subsequent improvement of the methodological apparatus of immunotoxicological studies of new drugs and assess the prospects of the development of new directions of immunopharmacology. The results of the information search for official data provided by manufacturers of medicines registered in the Russian Federation (guidelines for use, clinical recommendations, reports on biomedical and clinical research), as well as data from pharmacological monitoring of the safety of their use conducted by Roszdravnadzor of the Russian Federation are summarized.Guidelines for the use of 524 current international non-patent medicines, covering a total of more than 5000 generics in various forms, releases and dosages, were analyzed. The object of the search was data on undesirable side effects of medicines related to the human immune system.The analysis showed that the immunotoxic effect is described for 40,6% of 524 active pharmacological agents. Clinical and pathogenetic characteristics allowed us to identify several cluster groups that can be characterized as immunotoxic syndromes: allergic, immunosuppressive, lymphotropic, generalized cytokine reaction, reactivation of the immune response, and immune activation of oncogenesis. Molecular targets of immunotoxic action were determined based on the known mechanisms of action of the analyzed drugs.

Background and purpose Inhibition of the T- and B-cell interaction through the CD40/CD40 ligand (L) axis is a favourable approach for inflammatory disease treatment. Clinical studies of anti-CD40L molecules in autoimmune diseases have met challenges because of thromboembolic events and adverse haemostasis. VIB4920 (formerly MEDI4920) is a novel CD40L antagonist and Tn3 fusion protein designed to prevent adverse haemostasis and immunopharmacology. We evaluated the pharmacokinetics, activity and toxicity of VIB4920 in monkeys. Experimental approach Cynomolgus monkeys received i.v. or s.c. 5-300 mg·kg-1 VIB4920 or vehicle, once weekly for 1 month (Studies 1 and 2) or 28 weeks (Study 3). VIB4920 exposure and bioavailability were determined using pharmacokinetic analyses, and immune cell population changes via flow cytometry. Pharmacological activity was evaluated by measuring the animals' capacity to elicit an immune response to keyhole limpet haemocyanin (KLH) and tetanus toxoid (TT). Key results VIB4920 demonstrated linear pharmacokinetics at multiple doses. Lymphocyte, monocyte, cytotoxic T-cell and NK cell counts were not significantly different between treatment groups. B-cell counts reduced dose-dependently and the T-cell dependent antibody response to KLH was suppressed by VIB4920 dose-dependently. The recall response to TT was similar across treatment groups. No thromboembolic events or symptoms of immune system dysfunctionality were observed. Conclusions and implications VIB4920 demonstrated an acceptable safety profile in monkeys. VIB4920 showed favourable pharmacokinetics, dose-dependent inhibition of a neoantigen-specific immune response and no adverse effects on immune function following long-term use. Our data support the use of VIB4920 in clinical trials.

The immunopharmacological potential of alpha-fetoprotein (AFP) is closely related to its ability to modulate the activity of cells of the immune system. Due to the immunomodulating properties, as well as the transport function - the transfer of low molecular weight substances, AFP is used in the treatment of a number of pathological conditions. The first attempts to use the therapeutic potential of AFP are associated with the use of extracts of embryonic tissues, with the development of technology, purified AFP from abortive material was used as a drug, and in some cases, recombinant forms of AFP were used. These drugs are intended primarily for the treatment of autoimmune as well as cancer. The review presents data on known variants of AFP dosage forms. In general, the study of the mechanisms of action of AFP at the level of the immune system is currently an urgent issue.

The respiratory tract is constantly exposed to environmental aggressions including allergens, infections, and pollution. The immune system is pivotal to fight potential invaders and prevent the establishment of infections in the respiratory tract; however, excessive, misplaced or altered immune responses contribute to increase tissue damage, impair lung function, and exacerbate respiratory diseases. Indeed, asthma, chronic obstructive pulmonary disease (COPD) and cystic fibrosis are respiratory diseases that have uncontrolled inflammation as a unifying severity factor. In addition, the acute inflammatory responses in pneumonia must be controlled to avoid progression to acute respiratory distress syndrome, which can be lethal. In this regard, immunomodulatory therapies are promising pharmacological strategies to treat important respiratory pathologies. Understanding the pathogenesis of respiratory disorders have paved the way for the development of cytokine-directed therapies, allosteric or non-allosteric inhibitors of inflammatory receptors and enzymes, agonists of resolution of inflammation and other anti-inflammatory compounds. A multitude of studies have evaluated the safety, pharmacokinetics, and pharmacodynamics of several molecules that target inflammation in the respiratory tract, yet only few have been translated to clinical use. This article will summarize important aspects of asthma, COPD, cystic fibrosis, and pneumonia focusing on the role of immune responses in disease development and novel immunomodulatory therapies.

Immune system alterations have been implicated in various dopamine-related disorders, such as schizophrenia, bipolar disorder, and attention-deficit/hyperactivity disorder (ADHD). How immunity might be influenced by dopaminergic dysfunction and impact on clinically-relevant behaviors is still uncertain. We performed a peripheral and cerebral immunophenotyping in mice bearing dopaminergic alteration produced by genetic liability (hypofunction of the dopamine transporter DAT) and psychostimulant (amphetamine) administration. We found that DAT hypofunction influences immune tolerance by increasing functional Tregs and adrenomedullin levels in the thymus and spleen, while reducing microglia activation and infiltration of brain monocyte-derived macrophages (mo-MΦ). Remarkably, both DAT hypofunction and amphetamine treatment are associated with a weaker activation of the choroid plexus (CP) gateway. Conversely, amphetamine reactivated the CP in the setting of DAT hypofunction, paralleling its paradoxical ADHD-relevant behavioral effects. These findings add new knowledge on dopaminergic immunopharmacology and support the immunomodulation of CP functionality as a promising therapeutic strategy for neurodevelopmental and psychiatric disorders.

Rituximab (RTX) is an approved treatment for rheumatoid arthritis (RA) patients that do not respond adequately to disease-modifying antirheumatic drugs. However, different new concerns, such as efficacy, optimum dose, safety issues, prediction of response to RTX, and pregnancy outcomes have attracted a lot of attention. The PubMed database was systematically reviewed for the last published articles, new findings, and controversial issues regarding RTX therapy in RA using "Rheumatoid arthritis" AND "rituximab" keywords, last updated on June 18, 2019. From 1812 initial recorders, 162 studies met the criteria. Regarding the optimum dose, low-dose RTX therapy (2 × 500 mg) seems as effective as standard dose (2 × 1000 mg), safer, and more cost-effective. The most common reported safety challenges included de novo infections, false negative serologic tests of viral infections, reactivation of chronic infections, interfering with vaccination outcome, and development of de novo psoriasis. Other less reported side effects are infusion reactions, nervous system disorders, and gastrointestinal disorders. Lower exposure to other biologics, presence of some serological markers (e.g., anti-RF, anti-CCP, IL-33, ESR), specific variations in FCGR3A, FCGR2A, TGFβ1, IL6, IRF5, BAFF genes, and also EBV-positivity could be used to predict response to RTX. Although there is no evidence of the teratogenic effect of RTX, it is recommended that women do not expose themselves to RTX at least 6 months before the conception. Only a reversible reduction of B cell-count in the offspring may be the pregnancy-related outcome. Although RTX is an effective therapeutic option for RA, more studies on optimum doses, prevention of RTX-related side effects, prediction of RTX response, and safety during the pregnancy are required.

The beneficial effects promoted from the use of biomolecular substances into the formulations of personal care products are considered useful ingredients in cosmetic and therapeutic applications. Innovations in cosmetics are based on new bioactive formulations such as vitamins, oils, peptides, and protein hydrolysates. In skin care, the monomeric amino acids such as serine, threonine, alanine are common ingredients in cosmetics as they function as natural moisturizing factors which act as water-binding molecules. Amino acids and their salts e.g., arginine, glycine, etc. are also used as hair- and skin-conditioning agents in cosmetic formulations. The peptides are composed of short chain of amino acids and are used in cosmetics due to their numerous pathophysiological properties including anti-aging. There is growing interest in bioactive peptides in products for stimulating collagen and elastin synthesis in skin and improve surface healing. The main benefit of using proteins in cosmeceuticals is to improve the hydration of skin. Proteins increase the dehydration in the skin which helps to reduce wrinkles and improves the functions of the skin barrier. This review article describes the peptides, proteins that are most frequently used in cosmeceuticals and their potential benefits and practical use in cosmetic science and skincare.

Dopamine is well recognized as a neurotransmitter in the brain, and regulates critical functions in a variety of peripheral systems. Growing research has also shown that dopamine acts as an important regulator of immune function. Many immune cells express dopamine receptors and other dopamine related proteins, enabling them to actively respond to dopamine and suggesting that dopaminergic immunoregulation is an important part of proper immune function. A detailed understanding of the physiological concentrations of dopamine in specific regions of the human body, particularly in peripheral systems, is critical to the development of hypotheses and experiments examining the effects of physiologically relevant dopamine concentrations on immune cells. Unfortunately, the dopamine concentrations to which these immune cells would be exposed in different anatomical regions are not clear. To address this issue, this comprehensive review details the current information regarding concentrations of dopamine found in both the central nervous system and in many regions of the periphery. In addition, we discuss the immune cells present in each region, and how these could interact with dopamine in each compartment described. Finally, the review briefly addresses how changes in these dopamine concentrations could influence immune cell dysfunction in several disease states including Parkinson's disease, multiple sclerosis, rheumatoid arthritis, inflammatory bowel disease, as well as the collection of pathologies, cognitive and motor symptoms associated with HIV infection in the central nervous system, known as NeuroHIV. These data will improve our understanding of the interactions between the dopaminergic and immune systems during both homeostatic function and in disease, clarify the effects of existing dopaminergic drugs and promote the creation of new therapeutic strategies based on manipulating immune function through dopaminergic signaling. Graphical Abstract.

Objectives The cationic biopolymer chitosan (CH) has emerged as a promising candidate adjuvant due to its safety profile and immunostimulatory properties. The presence of endotoxin contamination in biomaterials is generally underappreciated and can generate misleading results. It is important to establish a convenient methodology to obtain large amounts of high quality chitosan nanoparticles for biomedical applications. Methods We developed an easy method to generate endotoxin-free chitosan and assessed its purity using the Limulus amebocyte lysate assay and by measuring dendritic cell activation. Key findings Purified chitosan-based formulations alone failed to induce production of the proinflammatory cytokines tumour necrosis factor alpha (TNF-α) and interleukin (IL)-6 in bone marrow-derived dendritic cells (BMDCs) generated from C57BL/6 mice, while maintaining its ability to promote IL-1β secretion in combination with the Toll-like receptor (TLR)-9 agonist, CpG. Moreover, BMDCs from C3H/HeN and TLR4-deficient mice, C3H/HeJ were stimulated with endotoxin-free chitosan-based formulations and no differences were observed in IL-6 and IL-1β secretion, excluding the involvement of TLR-4 in the immunomodulatory effects of chitosan. Conclusions The developed method provides simple guidelines for the production of endotoxin-free chitosan, ideal for biomedical applications.

Acute myeloid leukemia (AML) is an aggressive malignancy that requires rapid treatment with chemotherapies to reduce tumor burden. However, these chemotherapies can compromise lymphocyte function, thereby hindering normal anti-tumor immune responses and likely limiting the efficacy of subsequent immunotherapy. To better understand these negative impacts, we assessed the immunological effects of standard-of-care AML therapies on lymphocyte phenotype and function over time. When compared to healthy donors, untreated AML patients showed evidence of lymphocyte activation and exhaustion and had more prevalent CD57+NKG2C+ adaptive NK cells, which was independent of human cytomegalovirus (HCMV) status. HMA/venetoclax treatment resulted in a greater fraction of T cells with effector memory phenotype, inhibited IFN-γ secretion by CD8+ T cells, upregulated perforin expression in NK cells, downregulated PD-1 and 2B4 expression on CD4+ T cells, and stimulated Treg proliferation and CTLA-4 expression. Additionally, we showed increased expression of perforin and CD39 and enhanced IFN-γ production by T cells from pre-treatment blood samples of venetoclax-resistant AML patients. Our results provide insight into the lymphocyte status in previously untreated AML patients and the effects of standard-of-care treatments on their biology and functions. We also found novel pre-treatment characteristics of T cells that could potentially predict venetoclax resistance.

The demand for immunostimulants and immunomodulators in the pharmaceutical market is wide. They are used in the treatment of congenital, acquired immunodeficiency states, in the treatment of cancer patients, in the treatment of patients living in ecologically unfavorable regions of countries, etc. And modern immunopharmacology faces a global challenge in the development of targeted immunostimulants. Thus, the presented spectrum of immunostimulating drugs shows that each group of immunostimulants has a certain spectrum of side effects and synthetic drugs are a more promising group. Therefore, the screening of new promising immunostimulants of targeted synthetics is justified. To study the hemogram of peripheral blood, we used 48 adult female albino laboratory rats, and phenotyping of lymphocytes was carried out by the conventional method of indirect fluorescence using monoclonal antibodies. As a result, the blood leukogram changed towards the development of leukopenic processes. Compound 2 exhibited comparatively low lymphopoiesis-stimulating activity among the compounds studied. Compound 3 showed comparatively high lymphopoiesis-stimulating activity in the series of compounds 1, 2, and 3. The erythropoiesis-stimulating and thrombocytopoiesis-stimulating activity of compounds 1 and 3 was at the level of the reference drug methyluracil.

Coronavirus disease 2019 (COVID-19) affects the lungs but also the cardiovascular system, especially since the spike protein of the Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) uses the angiotensin-converting enzyme 2 (ACE2), existing on the membrane of many cell species, as a receptor. This connection has consequences on the renin-angiotensin system, arterial pressure and coagulation. An imbalance of blood pressure control, of coagulation and platelet functions, and of the kinin-kallicrein system may occur in natural disease but is also partly reflected in some possible negative consequences following vaccination. Knowledge of these mechanisms could help to identify the subjects most at risk of complications and to correctly carry out the causality assessment of adverse events.

Versión editorial disponible en: http://hdl.handle.net/10261/239002, Background: Antipsychotics suppress expression of inflammatory cytokines and inducible inflammatory enzymes. Elopiprazole (a phenylpiperazine antipsychotic drug in phase 1) has been characterized as a therapeutic drug to treat SARS-CoV-2 infection in a repurposing study. We aim to investigate the potential effects of aripiprazole (an FDA approved phenylpiperazine) on COVID19-related immunological parameters. Methods: Differential gene expression profiles of non-COVID versus COVID RNA-Seq samples (CRA002390 project in GSA database) and drug-naive patients with psychosis at baseline and after three months of aripiprazole treatment was identified. An integrative analysis between COVID and aripiprazole immunomodulatory antagonist effects was performed. Findings: 82 out the 377 genes (21.7%) with expression significantly altered by aripiprazole have also their expression altered in COVID-19 patients and in 93.9% of these genes their expression is reverted by aripiprazole. The number of common genes with expression altered in both analyses is significantly higher than expected (Fisher's Exact Test, two tail; P value=3.2e-11). 11 KEGG pathways were significantly enriched with genes with altered expression both in COVID-19 patients and aripiprazole medicated schizophrenia patients (P adj, This work was supported by: SAF2016-76046-R and SAF2013-46292-R (MINECO and FEDER) to B.C.F.

The brain responds to diabetic stress by inducing the inflammatory response. Under normal circumstances this process is tightly regulated. However, uncontrolled inflammatory responses lead to compromised function and eventual neurodegeneration. The microRNA (miR)-200 family, specifically miR-141, is differentially expressed in diseased states including cognitive decline, thereby triggering changes in downstream genes. We hypothesised that Metformin (MF) regulates the miR-141/protein phosphatase 2A (PP2A) axis, and associated NF-ĸB-mediated inflammasome expression in diabetic mice brain. Diabetes was induced by intraperitoneal injection of Streptozotocin (STZ), thereafter mice were treated with MF (20 mg/kg BW). Whole brain tissue was harvested for further analysis. In silico analysis showed that Sirt1 and PP2A are prediction targets of miR-141. Selected protein and gene expressions were established through western blotting and qPCR, respectively. Diabetic mice brain tissue demonstrated overexpression of miR-141 and related pro-inflammatory factors as well as decreased PP2A gene expression. MF was able to counteract this by regulating expression of miR-141, PP2A, and p-tau at Ser396 protein expressions. Further experimentation revealed MF's inhibitory action on the inflammasome system by regulating the expression of the upstream controller NLRP3, related cytokines and NF-κB signalling pathway. Collectively, we demonstrate that MF promotes neuroprotection in diabetic mice by dampening inflammatory responses through its inhibitory effects on various signalling pathways. Categories Inflammation and Immunopharmacology, Metabolic Disorders and Endocrinology, Neuropharmacology.

In this article we describe the use of pharmacological and genetic tools coupled with immunoblotting (Western blotting) and targeted mass spectrometry to quantify immune signaling and cell activation mediated by tyrosine kinases. Transfer of the ATP γ phosphate to a protein tyrosine residue activates signaling cascades regulating the differentiation, survival, and effector functions of all cells, with unique roles in immune antigen receptor, polarization, and other signaling pathways. Defining the substrates and scaffolding interactions of tyrosine kinases is critical for revealing and therapeutically manipulating mechanisms of immune regulation. Quantitative analysis of the amplitude and kinetics of these effects is becoming ever more accessible experimentally and increasingly important for predicting complex downstream effects of therapeutics and for building computational models. Secondarily, quantitative analysis is increasingly expected by reviewers and journal editors, and statistical analysis of biological replicates can bolster claims of experimental rigor and reproducibility. Here we outline methods for perturbing tyrosine kinase activity in cells and quantifying protein phosphorylation in lysates and immunoprecipitates. The immunoblotting techniques are a guide to probing the dynamics of protein abundance, protein–protein interactions, and changes in post‐translational modification. Immunoprecipitated protein complexes can also be subjected to targeted mass spectrometry to probe novel sites of modification and multiply modified or understudied proteins that cannot be resolved by immunoblotting. Together, these protocols form a framework for identifying the unique contributions of tyrosine kinases to cell activation and elucidating the mechanisms governing immune cell regulation in health and disease. © 2020 The Authors. Basic Protocol 1: Quantifying protein phosphorylation via immunoblotting and near‐infrared imaging Alternate Protocol: Visualizing immunoblots using chemiluminescence Basic Protocol 2: Enriching target proteins and isolation of protein complexes by immunoprecipitation Support Protocol: Covalent conjugation of antibodies to functionalized beads Basic Protocol 3: Quantifying proteins and post‐translational modifications by targeted mass spectrometry

Rheumatoid arthritis (RA) is a chronic immune-mediated inflammatory disorder characterized by the destruction of the joint and bone resorption. The production of pro-inflammatory cytokines and chemokines, dysregulated functions of three important subtypes of T helper (TH) cells including TH1, TH17, and regulator T (Treg) cells are major causes of the initiation and development of RA. Moreover, B cells as a source of the production of several autoantibodies play key roles in the pathogenesis of RA. The last decades have seen increasingly rapid advances in the field of immunopharmacology using natural origin compounds for the management of various inflammatory diseases. Curcumin, a main active polyphenol compound isolated from turmeric, curcuma longa, possesses a wide range of pharmacologic properties for the treatment of several diseases. This review comprehensively will assess beneficial immunomodulatory effects of curcumin on the production of pro-inflammatory cytokines and also dysregulated functions of immune cells including TH1, TH17, Treg, and B cells in RA. We also seek the clinical efficacy of curcumin for the treatment of RA in several recent clinical trials. In conclusion, curcumin has been found to ameliorate RA complications through modulating inflammatory and autoreactive responses in immune cells and synovial fibroblast cells via inhibiting the expression or function of pro-inflammatory mediators, such as nuclear factor-κB (NF-κB), activated protein-1 (AP-1), and mitogen-activated protein kinases (MAPKs). Of note, curcumin treatment without any adverse effects can attenuate the clinical symptoms of RA patients and, therefore, has therapeutic potential for the treatment of the diseases.

The main topic of this Thesis is to investigate the contribution of polyamines to defense in Arabidopsis thaliana and the requirement of callose deposition for full expression of effector-triggered immunity in autoimmune hybrids. Due to its accumulation during pathogen infection, I mainly focused on the polyamine putrescine. The interaction between polyamines, reactive oxygen species (ROS) production and salicylic acid pathway activation is also studied in the context of PAMP-triggered immunity (PTI) (Chapter 1) and systemic acquired resistance (SAR) (Chapter 2). The data support a role for putrescine as a priming agent contributing to resistance against pathogens, which can lead to practical applications in the development of PPP (plant protection products). In Chapter 3, I report the involvement of glucan synthase-like 2 and 10 (GSL2 and GSL10), two of the twelve callose biosynthesis genes, in the temperature-dependent immune hybrid incompatibility between natural accessions of Arabidopsis thaliana from North Europe (Ler) and Central Asia (Kas-2), which constitutes a model for the study of effector-triggered immunity (ETI). This work supports that PTI and ETI are not two separate branches of defense, but support each other through mutual potentiation. Overall, we provide evidence for the involvement of polyamines in defense signaling and callose biosynthesis in the establishment of ETI.

Discovery of vaccines is the most remarkable invention made in the field of science. These have shown to be successful when produced in animals but come with a lot of limitations. A striking invention was seen when the same vaccines could be produced in plants overcoming the disadvantages of animal vaccines. In this review, there is a detailed description underlying the need of producing vaccines in plants, why use plants as a host to produce vaccines, advantages of producing plant vaccines, existing technologies used in the production of plant vaccines and the strategies that can be employed in increasing its production. The review also focuses on how the small size of the plant genome can be utilized in order to bring about several genetic modifications making them resistant to different platforms in order to integrate the gene of interest. The review also shows lists of recombinant, oral vaccines produced through different technologies produced in plants.

Background: CD137 (4-1BB, TNFRSF9) is expressed on activated lymphocytes, and its clustering leads to agonism of the receptor resulting in lymphocyte proliferation and pro-inflammatory cytokine release. First-generation CD137 antibodies for cancer therapy were high affinity and enabled for FcγR engagement with either severe toxicity or weak activity limiting their therapeutic benefit. However, CD137 remains a promising target for bispecific antibodies designed to re-direct T cell activity to the tumor whilst limiting unwanted toxicities. We have rationally designed and developed a unique tetravalent bispecific antibody targeting CD137 and PD-L1 with reduced FcγR binding for safe and efficacious cancer therapy (Lakins MA et al. 2020). Methods: A CD137/PD-L1 bispecific mAb2 antibody (FS222) was generated by introducing a bivalent affinity-optimized CD137-binding Fcab into a human IgG1 bivalent PD-L1 mAb. LALA mutations were introduced to abrogate FcγR activity. To elucidate its mechanism of action, FS222's binding valency was assessed by chemically-crosslinked mass spectrometry mapping (XL/MS). Immune pharmacology models were also used to evaluate PD-L1 dependent FS222 agonism against variants with differing valency for each target. Results: FS222 was specifically designed to bind CD137 with moderate affinity, and without PD-L1-mediated crosslinking it does not cluster or activate the receptor on immune cells. When PD-L1 was bound by FS222, this lead to crosslinking and subsequent clustering and activation of CD137. This clustering was enabled by avid bivalent binding as demonstrated by tetravalent FS222 being biologically more active than variants with reduced valency. Moreover, XL/MS demonstrated all four binding sites were able to concurrently bind target antigens. FS222 was well tolerated in a GLP toxicity study up to the maximal dose (30mg/kg QW). It was pharmacologically active in peripheral blood and lymphoid tissues and had a predicted t½ of 286 hours in humans. A surrogate anti-mouse CD137/PD-L1 mAb2 significantly reduced tumor growth in a murine syngeneic model and this correlated with a significant survival benefit and T cell activation. Conclusions: FS222 was designed to be a potent anti-human CD137/PD-L1 tetravalent conditional agonist with a unique combination of high affinity PD-L1 binding and moderate monovalent affinity, and highly avid binding, to CD137 on activated T cells. A favorable safety profile and immunopharmacology was observed with FS222 in a primate GLP toxicity study. Tetravalent binding by FS222 was required for optimal activity in pharmacology studies. Preclinically, FS222 was well-tolerated with an effective mechanism of action. Citation Format: Matthew A. Lakins, Jose Munoz-Olaya, Christel Veyssier, Daniel Jones, Emma Goodman, Quincy Kaka, Jennifer Ofoedu, Robert Hughes, Daniel Gliddon, Michelle Morrow, Neil Brewis. FS222, a tetravalent bispecific antibody targeting CD137 and PD-L1, is designed for optimal CD137 interactions resulting in potent T cell activation without toxicity [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 1864.

Made available in DSpace on 2018-11-30T03:29:14Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-08-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) This report describes a model of host resistance for Sporothrix schenckii, an opportunistic fungi in immunosuppressed mice with cyclophosphamide (CY) to be used in studies of immunotoxicology and immunopharmacology. Two doses of CY were administered intraperitoneally: 200 mg/kg and a booster of 150 mg/kg at 9-day intervals. Three days after the first dose of CY the animals were infected subcutaneously with 1.8 x 10(8) yeast/ml (S. schenckii ATCC 16345). At 7 and 14 days post-infection, the animals were euthanized and analyzed the fungal load by unit forming colony count in the spleen and popliteal lymph nodes. The relative weight of thymus and spleen, splenic index, the frequency of T and B cells in spleen by flow cytometry, the hind paw inflammation index and cytokine (interleukin [IL]-17, IL-10, and interferon [IFN]-gamma) profile were measured. Histopathological studies of the spleen and the hind paw were also assessed. The immunosuppression status was confirmed at the evaluated days by reduction of relative weight of thymus, reduction of the splenic white pulp, the population of B and T lymphocytes, and the cytokine profile in the treated mice with CY in comparison with nontreated groups, associated to higher fungal load in hind paw and spleen in the infected mice. The described model reveals an increasing in susceptibility to infection and severity when associated with immunosuppression. This model can serve as a reference for studies of S. schenckii host resistance in pharmaceutical and toxicological studies. Sao Paulo State Univ, UNESP, Sch Pharmaceut Sci, Araraquara, SP, Brazil Sao Paulo State Univ, UNESP, Sch Pharmaceut Sci, Araraquara, SP, Brazil FAPESP: 2014/00914-5 FAPESP: 2015/04023-0

Antibody-drug conjugates (ADCs) represent an innovative therapeutic approach that provides novel treatment options and hope for patients with cancer. By coupling monoclonal antibodies (mAbs) to cytotoxic small-molecule payloads with a plasma-stable linker, ADCs offer the potential for increased drug specificity and fewer off-target effects than systemic chemotherapy. As evidence for the potential of these therapies, many new ADCs are in various stages of clinical development. Because their structure poses unique challenges to pharmacokinetic and pharmacodynamic characterization, it is critical to recognize the differences between ADCs and conventional chemotherapy in the design of ADC clinical development strategies. Although some properties may be determined mainly by either the mAb or the small-molecule portion, the behavior of these agents is not always predictable. Furthermore, because the absorption, distribution, metabolism, and excretion (ADME) of ADCs are influenced by all 3 of its components (mAb, linker, and payload), it is important to characterize the intact molecule, any target-mediated catabolic clearance of the mAb, and the ADME properties of the small-molecule payload. Here we describe key issues in the clinical development of ADCs, including considerations for designing first-in-human studies for ADCs. We discuss some difficulties of ADC pharmacokinetic characterization and current approaches to overcoming these challenges. Finally, we consider all aspects of clinical pharmacology assessment required during drug development, using examples from the literature to illustrate the discussion.

Antioxidants are agents which can modulate oxidant-antioxidant profile of body system by neutralizing pro-oxidant molecules. The current scientific knowledge on mechanisms of antioxidant activity of biomolecules was critically reviewed with a special emphasis on immunomodulation. The immuno-oxidative wreckage of animals in various disease conditions and the role of biomodulators in curbing the oxidative stress through immune pathways were analyzed. The critical role of immunomodulatory mechanisms in controlling oxidative damage was identified. Selection of antioxidant therapy considering the immunopharmacology of the drug as well as immunological basis of disease may reduce treatment failure and adverse health effects. Hence, it is suggested that future studies on antioxidants may focus on the immuno-oxidative pathobiology to better understand its clinical effects and effective disease management.

Searching for new natural drugs that are capable of targeting Th1 and Th17 may lead to development of more effective treatments for inflammatory and autoimmune diseases. Most of the natural drugs can be derived from plants that are used in traditional medicine and folk medicine. The aim of this systematic review is to identify and introduce plants or plant derivatives that are effective on inflammatory diseases by inhibiting Th1 and Th17 responses. To achieve this purpose, the search terms herb, herbal medicine, herbal drug, medicinal plant, phytochemical, traditional Chinese medicine, Ayurvedic medicine, natural compound, inflammation, inflammatory diseases, Th1, Th17, T helper 1 or T helper 17 were used separately in Title/Keywords/Abstract in Web of Science and PubMed databases. In articles investigating the effect of the medicinal plants and their derivatives in inhibiting Th1 and Th17 cells, the effects of eight extracts of the medicinal plants, 21 plant-based compounds and some of their derivatives, and eight drugs derived from the medicinal plants' compounds in inhibiting Th1 and Th17 cells were reviewed. The results showed that medicinal plants and their derivates are able to suppress Th17 and Th1 T cell functions as well as cytokine secretion and differentiation. The results can be used to produce herbal drugs that suppress Th, especially Th17, responses. Copyright © 2017 John Wiley & Sons, Ltd.

Recent advances in the bioengineering of monoclonal antibodies (mAbs) have revolutionized the treatment of several immunological and rheumatic diseases. mAbs exhibit high specificity and affinity, and are very effective targeting agents, associated with minimal off-target adverse effects. Of several relevant immunological diseases, rheumatoid arthritis was the condition initially treated with mAbs, with great success. Currently, many immunological disorders are targeted and successfully treated using such novel approaches; these include inflammatory bowel diseases, multiple sclerosis, lupus and psoriasis. Today, the efforts of researchers in basic immunology (with a long history) have borne fruit; bioengineered mAbs are employed in clinical practice. In this brief review, I will describe the current and emerging therapeutic mAbs and molecular targeted agents, and discuss the future of the field, especially from the viewpoint of pharmacology.

// Mandy Menzel 1 , Hamid Akbarshahi 1 , Ellen Tufvesson 2 , Carl Persson 3 , Leif Bjermer 2 , Lena Uller 1 1 Respiratory Immunopharmacology, Department of Experimental Medical Science, Lund University, 22184 Lund, Sweden 2 Respiratory Medicine and Allergology, Department of Clinical Sciences Lund, Lund University, 22184 Lund, Sweden 3 Division of Clinical Chemistry and Pharmacology, Department of Laboratory Medicine, Lund University, 22184 Lund, Sweden Correspondence to: Lena Uller, email: Lena.Uller@med.lu.se Keywords: asthma, rhinovirus, azithromycin, MDA5, IFNβ Received: February 08, 2017 Accepted: March 12, 2017 Published: March 18, 2017 ABSTRACT Deficient production of anti-viral interferons (IFNs) may be involved in causing viral-induced asthma exacerbations. Hence, drugs inducing lung IFN production would be warranted. Azithromycin may reduce asthma exacerbations but its modus operandi is unknown. Here, we investigated if azithromycin induces IFNβ expression in vitro in rhinovirus-infected bronchial epithelial cells from asthmatic donors and in vivo in our allergic inflammation-based mouse model of viral stimulus-induced asthma exacerbations. Azithromycin dose-dependently augmented viral-induced IFNβ expression in asthmatic, but not in healthy bronchial epithelial cells. The effect negatively correlated with viral load. Knockdown of MDA5 and RIG-I by siRNA showed involvement of MDA5 but not RIG-I in azithromycin’s IFN-inducing effects in vitro . In vivo azithromycin induced IFNβ protein, restoring a reduced lung IFN response exclusively in allergic exacerbating mice. This was associated with induction of interferon-stimulated genes and MDA5, but not RIG-I. We suggest that clinically relevant concentrations of azithromycin produce MDA5-dependent, anti-viral, IFN-inducing effects in bronchial epithelium distinctly from asthmatic donors. Similarly, azithromycin induced MDA5-associated IFN in virally stimulated lungs in vivo exclusively in allergic mice. Effects of azithromycin and MDA5-active drugs on viral-induced exacerbations deserve further research.

The present study reports the determination of optimum thawing temperature and holding period of the Mareks Disease (MD) cell associated vaccine after reconstitution by plaque assay. The highest titre of MD HVT+SB-1 bivalent vaccine was obtained at the thawing temperature of 35°C for 45 sec followed by 26°C for 45 sec, 20°C for 60 sec and 40°C at 45 sec; when the vaccine was held on ice after reconstitution. The difference in virus titre among 30, 90 and 120 min holding period at 26°C and 35°C thawing for 45 sec was significant (P≤0.001). However the variation in titre between 30 and 90 min holding period was not significant for thawing temperatures of 20°C and 40°C (P>0.05). No plaques were seen when the bivalent HVT+SB-1 vaccine was thawed at 45°C for 45 sec. The titre obtained at 40°C was extremely low after a holding period of 120 min. After thawing at 35°C for 45 sec and when the reconstituted vaccine was held at RT, there was a drastic reduction in virus titre by 120 min. The study revealed that thawing at 20°C and 40°C for 45 sec would severely lower the titre of HVT+SB-1 vaccine. The complete loss of virus titre was seen at thawing temperature of 45°C for 45 sec. The study reasserts the importance of maintaining the cold chain, appropriate thawing temperature and holding period after reconstitution of HVT+SB-1, a cell associated MD vaccine in maintaining the requisite PFU/dose of vaccine.

The IUPHAR/BPS Guide to PHARMACOLOGY (www.guidetopharmacology.org) is an open-access, expert-curated database of molecular interactions between ligands and their targets. We describe significant updates made over the seven releases during the last two years. The database is notably enhanced through the continued linking of relevant pharmacology with key immunological data types as part of the IUPHAR Guide to IMMUNOPHARMACOLOGY (www.guidetoimmunopharmacology.org) and by a major new extension, the IUPHAR/MMV Guide to Malaria PHARMACOLOGY (www.guidetomalariapharmacology.org). The latter has been constructed in partnership with the Medicines for Malaria Venture, an organization dedicated to identifying, developing and delivering new antimalarial therapies that are both effective and affordable. This is in response to the global challenge of over 200 million cases of malaria and 400 000 deaths worldwide, with the majority in the WHO Africa Region. It provides new pharmacological content, including molecular targets in the malaria parasite, interaction data for ligands with antimalarial activity, and establishes curation of data from screening assays, used routinely in antimalarial drug discovery, against the whole organism. A dedicated portal has been developed to provide quick and focused access to these new data.

Janus kinase (JAK) inhibitors have emerged as an effective class of therapies for various inflammatory diseases such as rheumatoid arthritis (RA). JAK inhibitors function intracellularly by modulating the catalytic activity of JAKs and disrupting the receptor‐mediated signaling of multiple cytokines and growth factors, including those with pro‐inflammatory activity. Understanding the inhibition profiles of different JAK inhibitors, based on the associated cytokine receptors and downstream inflammatory pathways affected, is important to identify the potential mechanisms for observed differences in efficacy and safety. This study applied an integrated modeling approach, using in vitro whole blood cytokine inhibition potencies and plasma pharmacokinetics, to determine JAK‐dependent cytokine receptor inhibition profiles, in the context of doses estimated to provide a similar clinical response in RA clinical trials. The calculated profiles of cytokine receptor inhibition for the JAK inhibitors tofacitinib, baricitinib, upadacitinib, and filgotinib and its metabolite, were generally similar when clinically efficacious doses for RA were considered. Only minor numerical differences in percentage cytokine receptor inhibition were observed, suggesting limited differentiation of these inhibitors based on JAK pharmacology, with each showing a differential selectivity for JAK1 heterodimer inhibition. Nevertheless, only robust clinical testing involving head‐to‐head studies will ultimately determine whether there are clinically meaningful differences between these JAK inhibitors. Furthermore, ongoing and future research into inhibitors with alternative JAK selectivity remains of clinical importance. Thus, all JAK inhibitors should be characterized via thorough preclinical, metabolic and pharmacological evaluation, adequate long‐term clinical data, and when available, real‐world experience.

Breast adenocarcinoma (BAC) in glandular tissue cells have excessive metastasis and invasion capability. The major challenges for the chemotherapy used for the management of BAC include chemoresistance and auto-immunosuppression in BAC. The 5-fluro uracil (5-FU) based therapy promotes the immune activation in BAC by targeting the regulatory T cells and myeloid-derived suppressor cells (MDSC). The beneficial effect of the combination of L-Arginine with 5-FU strives to be established in different pre-clinical and clinical conditions and explored in the scientific literature. L-Arginine induces NO production and potentiates the anticancer effect of 5-FU. NO-mediated signaling is regulated by nuclear factor erythroid 2-related factor 2 (NRF-2) mediated antioxidant response. NRF-2 mediated antioxidant mechanism always suppresses the formation of superoxide (O2 -) as well as other reactive oxygen species (ROS). Thus the utilization of NO by O2 - will be minimum in this combination therapy. The regulatory role of NRF-2 in regulation to Antioxidant Response Element (ARE) mediated cytoprotective gene expression in BAC remains unexplored. The present review summarizes the role of NRF-2 mediated antioxidant response on the synergistic antitumor effect of L-Arginine and 5-FU in BAC. This review brought new insight into the management of BAC and in the same context, a hypothesis is raised on the use of reduced glutathione (GSH) or N-Acetyl Cysteine as it may be an added adjuvant in the combination of 5- FU and L-Arginine for management of BAC.

Liposomal nanoparticles are a heterogeneous group of engineered drug carriers that have tremendous therapeutic potential in the treatment of cancer. They increase tumor drug delivery, significantly attenuate drug toxicity, and protect the drug from degradation. However, two decades after approval of the first nanoparticle-mediated anticancer drug, pegylated liposomal doxorubicin (Doxil), there has yet to be a major shift in cancer treatment paradigms. Only two anticancer nanoparticles are used in the first-line treatment of cancer patients, with all others relegated to the refractory or salvage setting. Herein, we discuss new insights into the mechanisms underlying in vivo interactions between liposomes and the tumor immunologic milieu, and the knowledge gaps that need to be addressed in order to realize the full clinical potential of cancer nanomedicines. We also discuss immunopharmacology insights from a parallel field, Cancer Immunotherapy, which have the potential to generate breakthroughs in Cancer Nanomedicine.

Increasing evidence supports that regulatory T cells (Tregs) within the tumor, tumor draining lymph nodes, ascites and peripheral blood of patients with cancer are associated with poor prognosis. Tregs are important mediators of active immune evasion in cancer. In this review, the potential mechanisms of Treg actions and the roles of Tregs specifically in the tumor microenvironment derived from three types of gynecological cancers, cervical, vulvar and ovarian, are described. The correlations between Tregs and clinical immunotherapeutic study outcomes are discussed. Successful modulation of Tregs would likely have significant impact on the effectiveness of immunotherapeutic treatments in cancer patients.

The medicinal properties of various nutritional components have been appreciated since ancient times. Hippocrates (460–377 B.C.), for example, stated: “Let medicine be thy food and food be thy medicine.” Tea brewed from various fruits, shrubs, and trees containing natural salicylates has been consumed for pain relief since the Stone Age. Also the origin of the most well-known painkiller found in almost every home—aspirin—is a willow bark tree extract (see Chap. 33). These examples illustrate how nature can provide the chemical structure for a pharmaceutical. It is now appreciated that 70% of current drugs have their origin in chemical compounds found in plants, fruits, and vegetables. Modern medicinal chemists are capable of isolating and identifying these active chemical compounds and then modifying them to yield compounds with increased activity and less side effects.

The role of the p38 signaling pathway in the innate and adaptive immune responses has been well documented, especially in inflammatory cytokine production by dendritic cells (DCs). However, whether the p38 signaling pathway affects the important antigen (Ag) presentation function of DCs remains largely unknown. In this study, we reported that the deletion of p38α resulted in an impaired cross-presentation ability of CD8(+) conventional DCs (cDCs) and a reduction in the direct presentation ability of CD8(−) cDCs ex vivo. Further study revealed that p38α had a crucial role in Ag processing by CD8(+) cDCs but did not affect the Ag uptake or co-stimulation of T cells. Moreover, p38α deficiency led to reduced cross-priming of T cells in vivo. The production of the IL-12p40 and IL-12p70 cytokines by p38α-deficient cDCs was also significantly reduced. Our study identified a new role for p38α in modulating the important antigen cross-presentation function of DCs.

N-Acetyl-l-cysteine (NAC) is a well-known medication, primarily used as a mucolytic agent in pulmonary disease. Recently, we have found that NAC possesses antipyretic properties. The aim of the present study was to investigate the mechanism by which NAC attenuates fever. The concentration of interleukin (IL)-10 and prostaglandin (PG) E2 were measured using ELISA kit in the supernatants aspirated after stimulation of peripheral blood mononuclear cells (PBMCs) with lipopolysaccharide (LPS, 1μg/mL) and NAC (10mM). The body temperature of the Wistar rats was measured using biotelemetry system. To inhibit endotoxic fever, NAC (200mg/kg; i.p.) was injected into the rats one hour prior to the LPS administration (50μg/kg; i.p.). The pre-treatment of LPS-stimulated PBMCs with NAC resulted in a significant decrease in PGE2 concentration in comparison to the cells treated with LPS alone (PGE2 level was 386.1±61.9pg/mL vs. 2078.9±157.9pg/mL, respectively, p0.001). Furthermore, in these cells we observed a significant increase in IL-10 level (142.1±2.62pg/mL in NAC+LPS stimulated cells vs. 54.4±0.6pg/mL in LPS stimulated cells, p0.001). The injection of anti-IL-10 antibody into the rats abolished antipyretic properties of NAC. Body temperature in animals treated with anti-IL-10+NAC/LPS was 38.28±0.12°C vs. 37.73±0.06°C in IgG+NAC/LPS rats (p0.001) and 38.31±0.20°C in NaCl/LPS-treated animals (n.s.). Based on these data, we conclude that NAC acts as an antipyretic via IL-10 stimulation. This finding provides a new insight into the immunopharmacology of NAC, and we believe that in a future it will contribute to the new and/or more accurate application of NAC in medicine.

// Sung Wook Son 1, * , Seok-Hyung Kim 2, 3, * , Eun-Yi Moon 4, * , Dong-Hoon Kim 5 , Suhkneung Pyo 6 , Sung Hee Um 1, 3 1 Department of Molecular Cell Biology, Samsung Biomedical Research Institute, Sungkyunkwan University School of Medicine, Gyeonggi-do, 16419, Korea 2 Department of Pathology, Samsung Medical Center, Sungkyunkwan University School of Medicine, Seoul, 06351, Korea 3 Department of Health Sciences and Technology, Samsung Advanced Institute for Health Sciences and Technology, Samsung Medical Center, Sungkyunkwan University, Seoul, 06351, Korea 4 Department of Bioscience and Biotechnology, Sejong University, Seoul, 05006, Korea 5 Department of Pharmacology, Korea University College of Medicine, Seoul, 02841, Korea 6 Division of Immunopharmacology, School of Pharmacy, Sungkyunkwan University, Gyeonggi-do, 16419, Korea * These authors have contributed equally to this work Correspondence to: Sung Hee Um, email: shum@skku.edu Keywords: V-ATPase subunit V1E1, esophageal squamous cell carcinoma (ESCC), cell motility, aerobic glycolysis, prognosis Received: April 05, 2016 Accepted: June 13, 2016 Published: June 30, 2016 ABSTRACT Vacuolar H +- ATPase (V-ATPase), a hetero-multimeric ATP-driven proton pump has recently emerged as a critical regulator of mTOR-induced amino acid sensing for cell growth. Although dysregulated activity of cell growth regulators is often associated with cancer, the prognostic significance and metabolic roles of V-ATPase in esophageal cancer progression remain unclear. Here, we show that high levels of V-ATPase subunit V1E1 (V-ATPase V1E1) were significantly associated with shortened disease-free survival in patients with esophageal squamous cell carcinoma (ESCC). Multivariate analysis identified the V-ATPase V1E1 as an independent adverse prognostic factor (hazard ratio;1.748, P = 0.018). In addition, depletion of V-ATPase V1E1 resulted in reduced cell motility, decreased glucose uptake, diminished levels of lactate, and decreased ATP production, as well as inhibition of glycolytic enzyme expression in TE8 esophageal cancer cells. Consistent with these results, the Cancer Genome Atlas (TCGA) data and Gene Set Enrichment Analysis (GSEA) showed a high frequency of copy number alterations of the V-ATPase V1E1 gene, and identified a correlation between levels of V-ATPase V1E1 mRNA and Pyruvate Kinase M2 (PKM2) in ESCC. High expression levels of both V-ATPase V1E1 and phosphorylated PKM2 (p-PKM2), a key player in cancer metabolism, were associated with poorer prognosis in ESCC. Collectively, our findings suggest that expression of the V-ATPase V1E1 has prognostic significance in ESCC, and is closely linked to migration, invasion, and aerobic glycolysis in esophageal cancer cells.

Natural killer (NK) cell education, a process for achieving functional maturation and self-tolerance, has been previously defined by the interaction between self-major histocompatibility complex class I (MHC-I) molecules and their specific inhibitory receptors. Over the past several years, growing evidence has highlighted the important roles of nonclassical MHC-I and non-MHC-I molecules in NK cell education. Herein, we review the current knowledge of NK cell education, with a particular focus on nonclassical MHC-I- and non-MHC-I-dependent education, and compare them with the classical MHC-I-dependent education theory. In addition, we update and extend this theory by presenting the 'Confining Model', discussing cis and trans characteristics, reassessing quantity and quality control, and elucidating the redundancy of NK cell education in tumor and virus infection.

Foot ulceration is one of the most debilitating complications associated with diabetes, but its cause remains poorly understood. Several studies have been undertaken to understand healing kinetics or find possible therapies to enhance healing. However, few studies have been directed at understanding the immunological alterations that could influence wound healing in diabetes. In this study, we analysed the T-cell receptor (TCR) repertoire diversity in TCR-αβ+ T cells. We also analysed the distribution and phenotype of T cells obtained from the peripheral blood of healthy controls and diabetic individuals with or without foot ulcers. Our results showed that diabetic individuals, especially those with foot ulcers, have a significantly lower naive T-cell number and a poorer TCR-Vβ repertoire diversity. We also showed that the reduced TCR-Vβ repertoire diversity in diabetic individuals was mainly owing to the accumulation of effector T cells, the major source of tumour necrosis factor-α production, which was even more pronounced in patients with acute foot ulceration. Moreover, the expression of several inflammatory chemokine receptors was significantly reduced in diabetic patients. In conclusion, effector T-cell accumulation and TCR repertoire diversity reduction appear to precede the development of foot ulcers. This finding may open new immunological therapeutic possibilities and provide a new prognostic tool in diabetic wound care.

Background Activating T cells via clustering tumour necrosis factor receptor superfamily (TNFRSF) has shown promise in cancer therapy. This has typically been achieved by crosslinking of targeting antibodies via Fc-gamma receptor (FcγR). TNFRSF member CD137 is expressed by activated lymphocytes, and its clustering results in lymphocyte proliferation and agonism. First generation CD137 antibodies induced liver immune pathology, causing clinical toxicity and death. It remains a promising target for bispecific antibodies designed to limit unwanted toxicities. Methods An anti-human CD137/PD-L1 mAb2 (FS222) was generated by introducing a CD137-binding specificity into a human IgG1 targeting PD-L1 mAb with reduced FcγR binding. Cell binding and in vitro activity was used to evaluate FS222 agonism conditional on PD-L1 crosslinking. An anti-mouse CD137/PD-L1 mAb2 was generated to assess anti-tumour activity, liver pharmacology and PK/PD in murine models. To predict the clinical toxicity and PK/PD profiles of FS222, studies were performed in non-human primates. Results FS222 is designed to bind human PD-L1 with sub-nanomolar affinity and crosslink to enable CD137 clustering and conditional agonism (low nM EC50). The toxicity, PK and PD of FS222 was tested in cynomolgus monkey. FS222 was well tolerated in a GLP toxicity study up to the maximal dose (30mg/kg QW) and showed no evidence of liver toxicity. The molecule has a t½ of 211h in monkeys, with treatment resulting in pharmacology in peripheral blood which included lymphocyte proliferation and increases in soluble receptors. The anti-mouse CD137/PD-L1 mAb2 significantly reduced tumor growth in mouse tumor models. This correlated with a significant survival benefit and increases in tumor and peripheral activated lymphocytes. An anti-mouse CD137 antibody (clone 3H3) showed crosslink-independent agonism and sustained increases in activated T cells in liver whereas the liver effects of the anti-mouse CD137/PD-L1 mAb2 were of similar magnitude but peaked earlier and resolved. Conclusions FS222 was observed to be a potent anti-human CD137/PD-L1 tetravalent conditional agonist. A favourable safety profile and immunopharmacology was observed with FS222 in a primate GLP toxicity study. FS222 did not show toxicity and the anti-mouse CD137/PD-L1 mAb2 had self-limiting immunopharmacology outside the tumor microenvironment, suggesting a well-tolerated and effective mechanism of action with a broad therapeutic window. Citation Format: Matthew A. Lakins, Alexander Koers, Raffaella Giambalvo, Robert Hughes, Sylwia Marshall, Mateusz Wydro, Cristian Gradinaru, Sarah Batey, Daniel Gliddon, Michelle Morrow, Neil Brewis. Clustering CD137 via cell-expressed PD-L1 crosslinking avoided Fc-mediated agonism and resulted in safe and potent conditional lymphocyte activation [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 4547.