Specific Immune Mechanisms Research Articles

Staphylococcus aureus Lipoteichoic Acid has a Dual Effect on Inflammatory Responses in Airway Epithelial Cells via the Transcription Factor NFκB

Staphylococcus aureus is a major human pathogen that causes a wide array of infections ranging in severity from mild skin infections to life-threatening diseases such as sepsis. S. aureus can cause severe inflammation whilst, at the same time, modulating specific innate and humoral immune defense mechanisms. Lipoteichoic acid (LTA) is a component of the cell wall of gram-positive bacteria that is abundantly released by S. aureus. We investigated the dose-dependent effects of LTA on inflammation in airway epithelial cells. Purified S. aureus LTA was applied to airway epithelial (NuLi-1) cells in a range of concentrations, followed by assessment of expression of NFκB p50 and p65/RelA via qPCR, immunofluorescence and western blot. Multiplex analysis of cell culture supernatants was carried out to evaluate cytokine levels. NFκB p50 mRNA varied according to the LTA dose (p = 0.013) whereas NFκB p65/RelA did not (p = 0.90). A dose-dependent increase in NFκB p50 mRNA expression was seen up to 10 μg/ml LTA treatment, before falling at the 100μg/ml treatment. Western blot analysis showed nuclear and cytoplasmic NFκB p50 and cytoplasmic NFκB p65 protein reduced with increasing LTA concentrations. Multiplex analysis demonstrated that cytokine levels increased with LTA dose, but then decreased at the highest LTA dose (100 μg/ml). This study identifies LTA as having a dual dose-dependent effect on immune activation at low dosages and ablation at high dosages, associated with altered expression levels of NFκB. It adds LTA to the extensive repertoire of possible immune modulators produced by S. aureus.

Neutrophil-Lymphocyte Ratio and the Platelet Parameters as Biomarkers of Atopic Dermatitis Severity in Children

Background: Atopic dermatitis (AD) is a chronic inflammatory skin disease with a specific immune and inflammatory mechanism. This study investigates inflammatory biomarkers and their correlation with disease severity. Objectives: The aim of this study is to determine the relationship between platelet parameters [(mean platelet volume (MPV), platelet distribution width (PDW)], the neutrophil-lymphocyte ratio (NLR), the platelet-lymphocyte ratio (PLR), and AD severity in children. Methods: In this retrospective study, we reviewed patients diagnosed with AD and a healthy control group at the department of pediatrics at Health Sciences in a university-affiliated hospital in Istanbul, Turkey, between January 2015 and December 2016. The study included 79 children with AD and 75 healthy controls. AD severity was graded using the Scoring Atopic Dermatitis (SCORAD) index. Complete blood count was measured, and NLR and PLR were calculated. NLR, PLR, MPV, PDW were compared between AD patients and healthy controls, and the correlations between these indexes and clinical characteristics were analyzed. Results: No significant difference was observed between patients and controls for MPV, NLR, and PLR (P = 0.708, P = 0.340, P = 0.179, respectively). In the AD group, PDW was lower than controls (17.39 ± 1.45, 18.04 ± 1.65, P = 0.006). In the severe AD group, MPV was higher (7.62 ± 1.81, 6.64 ± 1.16, P = 0.035) and PDW was lower than in the mild AD group (16.52 ± 1.49, 17.93 ± 1.44, P = 0.0001). Conclusions: Mean MPV and PDW levels are correlated with atopic dermatitis severity in children.

Alcohol-induced adipose tissue macrophage phenotypic switching is independent of myeloid Toll-like receptor 4 expression.

Alcoholic liver disease results from a combination of immune and metabolic pathogenic events. In addition to liver injury, chronic alcohol consumption also causes adipose tissue inflammation. The specific immune mechanisms that drive this process are unknown. Here, we sought to determine the role of the innate immune receptor Toll-like receptor 4 (TLR4) in alcohol-induced adipose tissue inflammation. Using a model of chronic, multiple-binge alcohol exposure, we showed that alcohol-mediated accumulation of proinflammatory adipose tissue macrophages was absent in global TLR4 knockout mice. Proinflammatory macrophage accumulation did not depend on macrophage TLR4 expression; LysMCre-driven deletion of Tlr4 from myeloid cells did not affect circulating endotoxin or the accumulation of M1 macrophages in adipose tissue following alcohol exposure. Proinflammatory cytokine/chemokine production in the adipose stromal vascular fraction also occurred independently of TLR4. Finally, the levels of other adipose immune cells, such as dendritic cells, neutrophils, B cells, and T cells, were modulated by chronic, multiple-binge alcohol and the presence of TLR4. Together, these data indicate that TLR4 expression on cells, other than myeloid cells, is important for the alcohol-induced increase in proinflammatory adipose tissue macrophages.

Identification and molecular characterization of exosome-like vesicles derived from the Taenia asiatica adult worm

Taenia asiatica is an important food-borne parasite that poses a threat to food-safety and animal husbandry hygine, yet little is known about its specific infection and immune escape mechanisms. Exosome-like vesicles have recently emerged as a regulator in the interactions between parasites and hosts, providing a new direction for research on infection of T. asiatica. In this experiment, exosome-like vesicles were collected from the excretory/secretory products of cultured T. asiatica and isolated by differential centrifugation. The purified vesicles, ranging from 30 to 150 nm in size, were identified as exosome-like vesicles by transmission electron microscope and Nanoparticle tracking analysis. Proteomics analysis identified 455 proteins in the exosome-like vesicles. Of these proteins, enzymes involved in metabolic processes were identified, including glyceraldehyde 3 phosphate dehydrogenase, fructose-1, 6-bisphosphate aldolase, cytosolic malate dehydrogenase, and enolase. The two most abundant proteins from proteomic analysis, 14-3-3 and enolase, were shown to be present in the exosome-like vesicles by immunogold labeling. High-throughput RNA sequencing yielded twenty known miRNAs present in exosome-like vesicle sRNA libraries. Nine of the miRNAs, including six known miRNAs (tas-miR-71, tas-miR-1, tas-miR-7, tas-miR-9, tas-miR-10, and tas-let-7) and three newly discovered miRNAs (tas-m0022-3p, tas-m0816-3p, tas-m0082-5p), were confirmed by RT-qPCR as present in T. asiatica adult worm extracts and secreted exosome-like vesicles in T. asiatica. Additionally, we demonstrated that exosome-like vesicles experimentally labeled with PKH67 were internalized by LoVo cells in vitro. These findings provide new insights into the interaction between tapeworms and hosts mediated by exosome-like vesicles.

Immune-mediated hookworm clearance and survival of a marine mammal decrease with warmer ocean temperatures.

Increases in ocean temperature are associated with changes in the distribution of fish stocks, and the foraging regimes and maternal attendance patterns of marine mammals. However, it is not well understood how these changes affect offspring health and survival. The maternal attendance patterns and immunity of South American fur seals were assessed in a rookery where hookworm disease is the main cause of pup mortality. Pups receiving higher levels of maternal attendance had a positive energy balance and a more reactive immune system. These pups were able to expel hookworms through a specific immune mediated mechanism and survived the infection. Maternal attendance was higher in years with low sea surface temperature, therefore, the mean hookworm burden and mortality increased with sea surface temperature over a 10-year period. We provide a mechanistic explanation regarding how changes in ocean temperature and maternal care affect infectious diseases dynamics in a marine mammal.

Immune Checkpoint Molecules, Personalized Immunotherapy, and Autoimmune Diabetes

Although significant progress has been made in understanding autoimmunity, no immunotherapy to effectively halt immune-mediated destruction of β cells in type 1 diabetes (T1D) is currently available. For successful immunotherapy it will be necessary to identify novel drug targets as well as robust immunologic biomarkers to predict disease heterogeneity and patient responsiveness. Inhibition of immune checkpoint mechanisms represents a novel and effective strategy in tumor immunotherapy. Because they are fundamental to rewiring immune circuits, the underlying mechanisms could be therapeutically enhanced and used as biomarkers in T1D. We examine here current knowledge of immune checkpoint molecules in T1D. One specific immune checkpoint mechanism, namely tryptophan metabolism, may meet the need for a valid drug target and robust biomarker in the quest for effective and personalized immunotherapy in T1D.

Inflammasome Priming Mediated via Toll-Like Receptors 2 and 4, Induces Th1-Like Regulatory T Cells in De Novo Autoimmune Hepatitis.

De novo autoimmune hepatitis (DAIH) is an important cause of late allograft dysfunction following liver transplantation, but its cause and underlying pathogenesis remains unclear. We sought to identify specific innate and adaptive immune mechanisms driving the pro-inflammatory cytokine secreting regulatory T cell (Treg) phenotype in DAIH and determine if modulation of these pathways could resolve the inflammatory milieu observed in the livers of patients with DAIH. Here, we demonstrate toll-like receptors (TLRs) 2- and 4-mediated inflammasome activation in CD14++ monocytes, a finding that is key to maintaining dysfunctional Tregs in patients with DAIH. Furthermore, silencing of TLR 2 and 4 in CD14++ monocytes prevented activation of the inflammasome and significantly decreased IFN-γ production by FOXP3+ Tregs. We also observed significantly increase in expression of tumor necrosis factor α-induced protein 3 (TNFAIP3), a negative regulator of the NLRP3 Inflammasome, in monocytes/macrophages of liver transplant subjects who have normal allograft function and do not have DAIH. TNFAIP3 expression was virtually absent in monocytes/macrophages of patients with DAIH. Our findings suggest that autoimmunity in DAIH is promoted by CD14++ monocytes predominantly through activation of inflammatory signaling pathways.

Effect of neoadjuvant chemotherapy on the immune microenvironment in non\u2013small cell lung carcinomas as determined by multiplex immunofluorescence and image analysis approaches

BackgroundThe clinical efficacy observed with inhibitors of programed cell death 1/programed cell death ligand 1 (PD-L1/PD-1) in cancer therapy has prompted studies to characterize the immune response in several tumor types, including lung cancer. However, the immunological profile of non–small cell lung carcinoma (NSCLC) treated with neoadjuvant chemotherapy (NCT) is not yet fully characterized, and it may be therapeutically important. The aim of this retrospective study was to characterize and quantify PD-L1/PD-1 expression and tumor-associated immune cells (TAICs) in surgically resected NSCLCs from patients who received NCT or did not receive NCT (non-NCT).MethodsWe analyzed immune markers in formalin-fixed, paraffin-embedded tumor tissues resected from 112 patients with stage II/III NSCLC, including 61 non-NCT (adenocarcinoma [ADC] = 33; squamous cell carcinoma [SCC] = 28) and 51 NCT (ADC = 31; SCC = 20). We used multiplex immunofluorescence to identify and quantify immune markers grouped into two 6-antibody panels: panel 1 included AE1/AE3, PD-L1, CD3, CD4, CD8, and CD68; panel 2 included AE1/AE3, PD1, granzyme B, FOXP3, CD45RO, and CD57.ResultsPD-L1 expression was higher (> overall median) in NCT cases (median, 19.53%) than in non-NCT cases (median, 1.55%; P = 0.022). Overall, density of TAICs was higher in NCT-NSCLCs than in non-NCT-NSCLCs. Densities of CD3+ cells in the tumor epithelial compartment were higher in NCT-ADCs and NCT-SCCs than in non-NCT-ADCs and non-NCT-SCCs (P = 0.043). Compared with non-NCT-SCCs, NCT-SCCs showed significantly higher densities of CD3 + CD4+ (P = 0.019) and PD-1+ (P < 0.001) cells in the tumor epithelial compartment. Density of CD68+ tumor-associated macrophages (TAMs) was higher in NCT-NSCLCs than in non-NCT-NSCLCs and was significantly higher in NCT-SCCs than in non-NCT-SCCs. In NCT-NSCLCs, higher levels of epithelial T lymphocytes (CD3 + CD4+) and epithelial and stromal TAMs (CD68+) were associated with better outcome in univariate and multivariate analyses.ConclusionsNCT-NSCLCs exhibited higher levels of PD-L1 expression and T-cell subset regulation than non-NCT-NSCLCs, suggesting that NCT activates specific immune response mechanisms in lung cancer. These results suggest the need for clinical trials and translational studies of combined chemotherapy and immunotherapy prior to surgical resection of locally advanced NSCLC.

TNFα and Radioresistant Stromal Cells Are Essential for Therapeutic Efficacy of Cyclic Dinucleotide STING Agonists in Nonimmunogenic Tumors.

The cGAS-STING cytosolic DNA sensing pathway may play an integral role in the initiation of antitumor immune responses. Studies evaluating the immunogenicity of various cyclic dinucleotide (CDN) STING agonists administered by intratumoral (i.t.) injection showed potent induction of inflammation, tumor necrosis, and, in some cases, durable tumor-specific adaptive immunity. However, the specific immune mechanisms underlying these responses remain incompletely defined. The majority of these studies have focused on the effect of CDNs on immune cells but have not conclusively interrogated the role of stromal cells in the acute rejection of the CDN-injected tumor. Here, we revealed a mechanism of STING agonist-mediated tumor response that relied on both stromal and immune cells to achieve tumor regression and clearance. Using knockout and bone marrow chimeric mice, we showed that although bone marrow-derived TNFα was necessary for CDN-induced necrosis, STING signaling in radioresistant stromal cells was also essential for CDN-mediated tumor rejection. These results provide evidence for crosstalk between stromal and hematopoietic cells during CDN-mediated tumor collapse after i.t. administration. These mechanistic insights may prove critical in the clinical development of STING agonists. Cancer Immunol Res; 6(4); 422-33. ©2018 AACR.

Effects of herbal nutraceuticals and/or zinc against Haemonchus contortus in lambs experimentally infected

BackgroundThe gastrointestinal parasitic nematode Haemonchus contortus is a pathogenic organism resistant to several anthelmintics. This study assessed the efficacy of a medicinal herbal mixture (Herbmix) and organic zinc, as an essential trace element for the proper functioning of both unspecific and specific immune defensive mechanisms, against experimental infections with H. contortus in lambs. All lambs were infected orally with approximately 5000 third-stage larvae of a strain of H. contortus susceptible to anthelmintics (MHco1). Twenty-four female lambs 3–4 months of age were divided into four groups: unsupplemented animals (control), animals supplemented with Herbmix (Hmix), animals supplemented with organic zinc (Zn) and animals supplemented with Herbmix and organic zinc (Hmix+Zn). Eggs per gram (EPG) of faeces were quantified 20, 28, 35, 42, 49, 56, 62 and 70 d post-infection and mean abomasal worm counts were assessed 70 d post-infection. Samples of blood were collected from each animal 7, 35, 49 and 70 d post-infection.ResultsQuantitative analyses of the bioactive compounds in Herbmix identified three main groups: flavonoids (9964.7 μg/g), diterpenes (4886.1 μg/g) and phenolic acids (3549.2 μg/g). Egg counts in the lambs treated with Hmix, Zn and Hmix+Zn decreased after 49 d. The EPGs in the Zn and Hmix+Zn groups were significantly lower on day 56 (P < 0.05 and P < 0.01, respectively), and the EPGs and mean worm counts were significantly lower on day 70 in all supplemented groups (P < 0.05 and P < 0.01). Hemograms of complete red blood cells of each animal identified clinical signs of haemonchosis after day 35. Serum calprotectin concentrations and IgA levels were significantly affected by treatment. The treatment influenced serum malondialdehyde concentrations (P < 0.05) and sulfhydryl groups (P < 0.01) of antioxidant status. The mineral status was unaltered in all lambs.ConclusionA direct anthelmintic impact on the viability of nematodes was not fully demonstrated, but the treatments with herbal nutraceuticals and zinc likely indirectly contributed to the increase in the resistance of the lambs to nematode infection.

Immune Response to Opisthorchis viverrini Infection and Its Role in Pathology.

Human liver fluke infection caused by Opisthorchis viverrini is a major public health problem in Mekong countries such as Thailand, Laos, Cambodia, Vietnam, and Myanmar with over 10 million infected through consumption of fish containing infective metacercariae. With no tissue migration phase and living entirely within the larger secondary (intrahepatic) bile ducts, liver flukes are only exposed to a biliarymucosal immune response, while their excretory and secretory products also stimulate chronic inflammation of biliary epithelium. Neither mucosal nor tissue immune responses appear to cause parasite death or protect against newly established flukes, as evidenced by the persistence of infection for decades in the body and rapid reinfection following treatment. Experimental studies suggest that specific immune suppressive mechanisms may promote parasite persistence, therefore allowing continued secretion of parasite products that damage the biliary epithelium, both directly through mechanical damage and mitogenicity and through innate and adaptive inflammatory responses. Chronic infection is associated with several hepatobiliary diseases, specifically gallbladder and bile duct inflammation (cholecystitis and cholangitis), periductal fibrosis, and cholangiocarcinoma, the fatal bile duct cancer. Various studies have linked the chronic immune response to parasite antigens to both fibrosis and many steps in the carcinogenic process. Here, we review research-based understandings of the basic immune response to liver fluke infection and its roles in host protection and immunopathogenesis from available literature and also from recent studies conducted by the authors.

Deciphering sex differences in the immune system and depression.

Certain mood disorders and autoimmune diseases are predominately female diseases but we do not know why. Here, we explore the relationship between depression and the immune system from a sex-based perspective. This review characterizes sex differences in the immune system in health and disease. We explore the contribution of gonadal and stress hormones to immune function at the cellular and molecular level in the brain and body. We propose hormonal and genetic sex specific immune mechanisms that may contribute to the etiology of mood disorders.

Abstract 2934: Neoadjuvant chemotherapy influence changes of the immune response in non-small cell lung carcinomas immune response in non-small cell lung carcinomas

Abstract Background: The clinical efficacy observed with PD-1/PD-L1 inhibitors in non-small cell lung carcinoma (NSCLC) has prompted to characterize the immune response in lung tumors treated with chemotherapy. The aim of this study was to determine the changes of the immune microenvironment in surgically resected NSCLCs from patients who received and did not receive neo-adjuvant chemotherapy. Methods: We studied formalin-fixed and paraffin embedded (FFPE) tumor tissues from 112 stage II/III resected NSCLC, including 61 chemotherapy-naïve (adenocarcinoma, ADC=33; squamous cell carcinoma, SCC=28) and 51 chemotherapy-treated (ADC=31; SCC=20) tumors. mIF was performed using the Opal 7-color fIHC Kit™, scanning in the Vectra™ multispectral microscope and analyzed using the inForm™ software (Perkin Elmer, Waltham, MA). The markers studied were grouped in two 6-antibody panels: Panel 1, AE1/AE3 pancytokeratins, PD-L1 (clone E1L3N), CD3, CD4, CD8 and CD68; and Panel 2, AE1/AE3, PD1, Granzyme B, FOXP3, CD45RO and CD57. Results: Positive PD-L1 expression (&amp;gt;5%) in malignant cells (MCs) was detected in 48% (n=53/112) of NSCLCs. Overall, chemotherapy-treated tumors showed significantly higher percentages of MCs expressing PD-L1 (median, 19.52%) than chemotherapy-naïve cases (median, 1.54%; P=0.008). Higher densities of lymphocytes expressing CD3+ (P=0.021), CD4+ (P=0.05), CD57 (P&amp;lt;0.001), CD45RO+ (P=0.019), and PD-1 (P=0.016) were detected in chemotherapy-treated NSCLCs compared with chemo-naïve tumors. In contrast, lower densities of FOXP3+ regulatory T cells and CD68+ macrophages but not statistical significant were detected in chemotherapy-naïve tumors when compared with chemotherapy-treated cases. Following chemotherapy ADCs exhibited significantly higher levels of CD57+ (P=0.008) and high density of PD-1 expressing by CD45RO+ cells (P=0.016) than chemotherapy-naïve tumors. Chemotherapy-treated SCCs demonstrated higher density of TAMs CD68+, CD57+,CD45RO+ and PD-1 cells than chemotherapy-naïve tumors (P&amp;lt;0.05). In chemotherapy-treated cancers, lower levels of CD4+ helper T cells was associated with worse overall survival (OS; P=0.04) in univariate analysis. In chemotherapy-treated ADC patients, lower levels of CD68-positive (P=0.010) and higher levels of FOXP3-positive cells correlated with worse OS (P=0.044). Conclusions: Neo-adjuvant chemotherapy-treated NSCLCs exhibited higher levels of PD-L1 expression and T cell subsets regulation compared to chemotherapy-naïve tumors, suggesting that chemotherapy activates specific immune response mechanisms in lung cancer. These results provide an inclination towards the design of clinical trials combining neo-adjuvant chemotherapy and immunotherapy prior to surgical resection of locally advanced NSCLC. (Supported by CPRIT MIRA and UT Lung SPORE grants, and MD Anderson Moon Shot Program). Citation Format: Edwin Roger Cuentas, Carmen Behrens, Jaime Rodriguez-Canales, Mei Jiang, Apar Pataer, Arelene Correa, Stephen Swisher, Boris Sepesi, Annikka Weissferdt, Neda Kalhor, Jiexin Zhang, Jack Lee, John Heymach, Cesar Moran, Jianjun Zhang, Don Gibbons, Ignacio Wistuba. Neoadjuvant chemotherapy influence changes of the immune response in non-small cell lung carcinomas immune response in non-small cell lung carcinomas [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 2934. doi:10.1158/1538-7445.AM2017-2934

Elevated Plasma Growth Arrest-Specific 6 Protein Levels Are Associated with the Severity of Disease During Hantaan Virus Infection in Humans.

Hemorrhagic fever with renal syndrome (HFRS) is caused by Hantaan virus (HTNV) and Seoul virus infection in Asia. The clinical manifestation of HFRS is characterized by the rapid loss of renal function (acute kidney injury) and thrombocytopenia. The specific immune mechanisms that cause thrombocytopenia in HFRS are not well described. The growth arrest-specific 6 (Gas6) protein and TAM (Tyro3, Axl and Mer) receptors have been recently shown to play prominent roles in immune regulation, and high plasma levels of Gas6 may predict the severity of diseases. The association of TAM receptors with several autoimmune diseases has been investigated, although the relationship between TAM receptors and these diseases remains unclear in HFRS. Therefore, the aim of this study was to evaluate the clinical significance of Gas6 and TAM receptor expression in HFRS. The concentrations of Gas6 in the plasma from 144 patients and the expression of TAM receptors on monocytes from 117 patients were quantified. The relationship between Gas6 levels and disease course, severity, and clinical parameters was analyzed. We first found that the plasma Gas6 levels were significantly higher in HFRS patients, whereby they were positively correlated with white blood cell counts and negatively correlated with platelet counts. The expression of Tyro3 was increased on monocytes in HFRS patients compared with that in controls. Taken together, our data indicate that elevated plasma Gas6 levels is associated with the severity of disease during HTNV infection in humans, suggesting that Gas6 may play an important role by binding with Tyro3 on monocytes, which will be assessed in future studies.

Phenotypic characterization and anticancer capacity of CD8+ cytokine-induced killer cells after antigen-induced expansion

Cytokine-induced killer cells (CIK) have been used in clinic for adoptive immunotherapy in a variety of malignant tumors and have improved the prognosis of cancer patients. However, there are individual differences in the CIK cell preparations including the obvious differences in the ratio of effector CIK cells among different cancer patients. Infusion of such heterogeneous immune cell preparation is an important factor that would affect the therapeutic efficacy. We report here the enrichment and expansion of CD8+ cells from CIK cells cultured for one week using magnetic activated cell sorting (MACS). These enriched CD8+ CIK cells expressed T cell marker CD3 and antigen recognition receptor NKG2D. Phenotypic analysis showed that CD8+ CIK cells contained 32.4% of CD3+ CD56+ natural killer (NK)-like T cells, 23.6% of CD45RO+ CD28+, and 50.5% of CD45RA+ CD27+ memory T cells. In vitro cytotoxic activity assay demonstrated that the enriched CD8+ CIK cells had significant cytotoxic activity against K562 cells and five ovarian cancer cell lines. Intriguingly, CD8+ CIK cells had strong cytotoxic activity against OVCAR3 cells that has weak binding capability to NKG2D. Flow cytometry and quantitative RT-PCR analysis revealed that OVCAR3 cells expressed HLA-I and OCT4 and Sox2, suggesting that CD8+ CIK cells recognize surface antigen via specific T cell receptor and effectively kill the target cells. The results suggest that transplantation of such in vitro enriched and expanded OCT4-specific CD8+ CIK cells may improve the specific immune defense mechanism against cancer stem cells, providing a novel avenue of cancer stem cell targeted immunotherapy for clinical treatment of ovarian cancer.

Abstract PR11: eFT508: An oral, potent and highly selective inhibitor of MNK1 and MNK2, promotes anti-tumor immunity as a monotherapy and in combination with immune checkpoint blockade

Abstract Purpose: This study was designed to evaluate the potential of eFT508 to selectively regulate key immune signaling pathways and enhance anti-tumor immunity as a monotherapy or in combination with checkpoint blockade in immunocompetent syngeneic cancer models. Methods: eFT508 and its effect on mRNA translation, effector protein production, immune cell signaling and tumor infiltrating lymphocytes was evaluated in vitro using normal human T cells and in vivo utilizing immunocompetent syngeneic models. The mechanism of translational regulation of specific target genes was further evaluated in these model systems. Results: Dysregulated translation of messenger RNA (mRNA) plays a role in the pathogenesis of multiple solid tumors and hematological malignancies. MNK1 and MNK2 integrate signals from several oncogenic and immune signaling pathways (including RAS, p38 and toll-like receptors) by phosphorylating eukaryotic initiation factor 4E (eIF4E) and other key effector proteins including hnRNPA1 and PSF. Phosphorylation of these RNA-binding proteins by MNK1 and MNK2 selectively regulates the stability and translation of a subset of cellular mRNA that control tumor/stromal cell signaling and the tumor microenvironment. eFT508 inhibits both MNK1 and MNK2 through a reversible, ATP-competitive mechanism of action with an IC50 of 2 and 1 nM against MNK1 and MNK2 respectively. eFT508 is highly selective (≥100-fold) for MNK1 and MNK2 relative to over 400 other protein and lipid kinases. Ribosome profiling has demonstrated that inhibition of MNK1 and MNK2 by eFT508 selectively regulates the translational efficiency and mRNA stability of a subset of genes that include inflammatory cytokines/chemokines, regulators of reactive oxygen species (ROS), and effectors of anti-tumor immune response. Given the importance of both RAS signaling and translational control to immune cell function the immunological effect of eFT508 was evaluated in both normal human T cells in vitro and immunocompetent syngeneic cancer models in vivo. eFT508 treatment of normal donor T cells has no deleterious effect on CD3/CD28 activation of IL-2 production, T cell proliferation or on T cell viability. However, eFT508 selectively down regulates the induction of IL-10 and specific immune checkpoint mechanisms. The effect of eFT508 on IL-10 protein production corresponded with reduced mRNA stability. The in vivo antitumor effect of eFT508 was assessed in the CT26 BALB/C syngeneic tumor model. CT26 mouse tumor cell proliferation and survival are insensitive to eFT508 in vitro. In vivo, daily oral treatment with 1 mg/kg eFT508 results in significant anti-tumor activity and establishment of immune memory. In addition, combination of daily oral treatment of 1 mg/kg eFT508 with either anti-PD-1 or anti-PD-L1 monoclonal antibodies increases the number of responder animals and results in synergistic activity that corresponds to the modulation of tumor infiltrating lymphocyte populations. Conclusions: eFT508 is a selective, orally bioavailable small molecule inhibitor of MNK1 and MNK2 that can decrease the production of key immune checkpoint regulators and immunosuppressive cytokines. This novel mechanism of action triggers anti-tumor immune response in immunocompetent syngeneic animal models as a monotherapy and in combination with established immune checkpoint antibodies. eFT508 is currently under evaluation in two phase I clinical trials for patients with advanced solid tumors and patients with advanced lymphoma respectively. These findings support further clinical evaluation of eFT508 in combination with checkpoint blockade. This abstract is also being presented as Poster B29. Citation Format: Kevin R. Webster, Vikas K. Goel, Jocelyn Staunton, Ivy NJ Hung, Gregory S. Parker, Craig R. Stumpf, Jolene Molter, Gary G. Chiang, Christopher J. Wegerski, Samuel Sperry, Joan Chen, Vera Huang, Peggy A. Thompson, Chinh Tran, Justin T. Ernst, Stephen E. Webber, Paul A. Sprengeler, Siegfried H. Reich. eFT508: An oral, potent and highly selective inhibitor of MNK1 and MNK2, promotes anti-tumor immunity as a monotherapy and in combination with immune checkpoint blockade. [abstract]. In: Proceedings of the AACR Special Conference on Translational Control of Cancer: A New Frontier in Cancer Biology and Therapy; 2016 Oct 27-30; San Francisco, CA. Philadelphia (PA): AACR; Cancer Res 2017;77(6 Suppl):Abstract nr PR11.

The establishment of cow's milk protein allergy in infants is related with a deficit of regulatory T cells (Treg) and vitamin D.

Cow's milk protein allergy (CMPA) is the most common food allergy in infants. However, little is known about which specific immune mechanisms are related with the CMPA onset. The objective was to investigate which immune alterations constitute differential factors between allergy and tolerance, and hence could be implicated in the CMPA establishment in infants. An extensive analysis of immune subsets, including Treg and cytokine-secreting cells was performed in blood samples from 28 infants younger than 9 mo obtained 1-4 d after the first adverse reaction to milk. Less than 4 d after first allergic reaction, infants who developed CMPA had decreased Treg counts and increased frequency of IL4-secreting CD4 T cells compared to controls. The deficit of Tregs was correlated with decreased serum levels of vitamin D. Values of Tregs, IL4-secreting cells and vitamin D were good predictors of CMPA diagnosis. Basal vitamin D levels in CMPA infants also predicted those CMPA patients developing spontaneous tolerance in the first year. Establishment of CMPA in infants was related with lower Treg and vitamin D levels. These immune alterations would be crucial factors behind the CMPA establishment and they could constitute a therapeutic target for treatment of CMPA.

Moving Biomarkers toward Clinical Implementation in Kidney Transplantation.

Long-term kidney transplant outcomes remain suboptimal, delineating an unmet medical need. Although current immunosuppressive therapy in kidney transplant recipients is effective, dosing is conventionally adjusted empirically on the basis of time after transplant or altered in response to detection of kidney dysfunction, histologic evidence of allograft damage, or infection. Such strategies tend to detect allograft rejection after significant injury has already occurred, fail to detect chronic subclinical inflammation that can negatively affect graft survival, and ignore specific risks and immune mechanisms that differentially contribute to allograft damage among transplant recipients. Assays and biomarkers that reliably quantify and/or predict the risk of allograft injury have the potential to overcome these deficits and thereby, aid clinicians in optimizing immunosuppressive regimens. Herein, we review the data on candidate biomarkers that we contend have the highest potential to become clinically useful surrogates in kidney transplant recipients, including functional T cell assays, urinary gene and protein assays, peripheral blood cell gene expression profiles, and allograft gene expression profiles. We identify barriers to clinical biomarker adoption in the transplant field and suggest strategies for moving biomarker-based individualization of transplant care from a research hypothesis to clinical implementation.

A real-time immune planning algorithm incorporating a specific immune mechanism for multi-robots in complex environments

To solve the real-time path planning of multi-robots in complex environments, a new immune planning algorithm incorporating a specific immune mechanism is presented. In the immune planning algorithm incorporating a specific immune mechanism, a new coding format for an antibody is first defined according to the impact of the obstacle distribution on the obstacle avoidance behaviors of multi-robots. Then, a new robot immune dynamic model for antibody selection is designed in terms of different impacts of obstacles and targets on robot behaviors. Finally, aiming at the local minimum problem in complex environments and inspired by the specific immune mechanism, a series of appropriate avoidance behaviors are selected through the calculation of a specific immune mechanism to help robots walk out of local minima. In addition, to solve deadlock situations, a learning strategy for the antibody concentration is presented. Compared with four related immune planning algorithms—an improved artificial potential field, a rapidly exploring random tree algorithm, a D* algorithm and a A* algorithm—the simulation results in four static environments show that the paths planned by immune planning algorithm incorporating a specific immune mechanism are the shortest and the path smoothness is generally the highest, which shows its strong planning capability in multi-obstacle environments. The simulation result in a dynamic environment with local minima shows that the immune planning algorithm incorporating a specific immune mechanism has strong planning ability in dynamic obstacle avoidance and in escaping from local minima. Additionally, an experiment in a multi-robot environment shows that two robots can not only avoid static obstacles but also avoid dynamic obstacles, which further supports the validity of the proposed immune planning algorithm incorporating a specific immune mechanism for multi-robots in real environments.

The clinical immunologic characteristic and laboratory diagnostic of bronchial asthma under pecilomycosis.

The actuality of the problem is is conditioned by detection of infection of patients' blood with fungi genus paecilomyces in the form of carriage or activation of pecilomycosis infection with clinical manifestation of allergic diseases and bronchial asthma. The purpose of study is to analyze clinical and immunologic characteristics of bronchial asthma under pecilomycosis and to develop new approaches to its laboratory diagnostic in conditions of the primary health care unit. The complex examination of patients that included collection of allergy anamnesis, physical examination, application of laboratory, cytology, immunology and microbiology analysis was carried out. The diagnosis of pecilomycosis was established at the basis of express-diagnostic using direct count of mature spherules and yeast-like cells of fungi genus Paecilomyces in blood sample, detection of specific IgE- and IgG antibodies to allergen of fungus Paecilomyces spp. in blood of patients and cultural analysis. The clinical immunological characteristics of atopic and endogenous (non-atopic) forms of bronchial asthma in 143 patients were examined. The activation of pecilomycosis was established in 120 (83.9%) patients: the content of mature spherules of fungus genus Paecilomyces in blood varied from 8 000 to 55 000 (in average 17 500±755) in 1 mkl at the standard 1000-6000 (in average 3417±433) in 1 mkl. The specific IgE- and IgG antibodies to allergen of fungus Paecilomyces spр. were detected (57% and 81% correspondingly). The development of bronchial asthma is induced by activation of pecilomycosis in blood: it occurred against the background of acute respiratory viral infection in 74 patients (62%), seasonal and age fluctuations of immune status in 34 patients (28%), under stress impact in 12 patients (10%). The immune response to infection with fungus genus Paecilomyces is conditioned by mechanisms of non-specific resistance and specific immunity. The eosinophils and natural killers are involved into sanation of blood from fungal cells of non-phagocyting sizes using antibodies-mediators of specific IgE and IgG antibodies to allergen of fungus Paecilomyces as effector cells in reaction of antibody-dpendent and cell mediated cytotoxicity. The phagocytosis of small fungal forms was detected: endospores by monocytes, neutrophils, eosinophils and cytolysis of fungal cells by erythrocytes. The obtained data permits to differentiate carriage or activation of pecilomycosis in blood of patients with bronchial asthma, to mark out individuals with atopic or non-atopic profile of immune reaction determining specificity of their immune defense from agent of fungal infection and clinical picture of disease. The algorithm is proposed concerning diagnostic of bronchial asthma of pecilomycosis etiology in conditions of primary health care unit.