Higher Levels Of IL-4 Research Articles

Interleukin 22 Signaling Regulates Acinar Cell Plasticity to Promote Pancreatic Tumor Development in Mice

Pancreatic ductal adenocarcinoma (PDAC) is an aggressive malignancy that invades surrounding structures and metastasizes rapidly. Although inflammation is associated with tumor formation and progression, little is known about the mechanisms of this connection. We investigate the effects of interleukin (IL) 22 in the development of pancreatic tumors in mice. We performed studies with Pdx1-Cre;LSL-KrasG12D;Trp53+/-;Rosa26EYFP/+ (PKCY) mice, which develop pancreatic tumors, and PKCY mice with disruption of IL22 (PKCY Il22-/-mice). Pancreata were collectedat different stages of tumor development and analyzed by immunohistochemistry, immunoblotting, real-time polymerase chain reaction, and flow cytometry. Some mice were given cerulean to induce pancreatitis. Pancreatic cancer cell lines (PD2560) were orthotopically injected into C57BL/6 mice or Il22-/-mice, and tumor development was monitored. Pancreatic cells were injected into the tail veins of mice, and lung metastases were quantified. Acini were collected from C57BL/6 mice and resected human pancreata and were cultured. Cell lines and acini cultures were incubated with IL22 and pharmacologic inhibitors, and protein levels were knocked down with small hairpin RNAs. We performed immunohistochemical analyses of 26 PDACs and 5 nonneoplastic pancreas specimens. We observed increased expression of IL22 and the IL22 receptor (IL22R) in the pancreas compared with other tissues in mice; IL22 increased with pancreatitis and tumorigenesis. Flow cytometry indicated that the IL22 was produced primarily by T-helper 22 cells. PKCY Il22-/-mice did not develop precancerous lesions or pancreatic tumors. The addition of IL22 to cultured acinar cells increased their expression of markers of ductal metaplasia; these effects of IL22 were prevented with inhibitors of Janus kinase signaling to signal transducer and activator of transcription (STAT) (ruxolitinib) or mitogen-activated protein kinase kinase (MEK) (trametinib) and with STAT3 knockdown. Pancreatic cells injected into Il22-/- mice formed smaller tumors than those injected into C57BL/6. Incubation of IL22R-expressing PDAC cells with IL22 promoted spheroid formation and invasive activity, resulting in increased expression of stem-associated transcription factors (GATA4, SOX2, SOX17, and NANOG), and increased markers of the epithelial-mesenchymal transition (CDH1, SNAI2, TWIST1, and beta catenin); ruxolitinib blocked these effects. Human PDAC tissues had higher levels of IL22, phosphorylated STAT3, and markers of the epithelial-mesenchymal transition than nonneoplastic tissues. An increased level of STAT3 in IL22R-positive cells was associated with shorter survival times of patients. We found levels of IL22 to be increased during pancreatitis and pancreatic tumor development and to be required for tumor development and progression in mice. IL22 promotes acinar to ductal metaplasia, stem cell features, and increased expression of markers of the epithelial-mesenchymal transition; inhibitors of STAT3 block these effects. Increased expression of IL22 by PDACs is associated with reduced survival times.

Application of the Zebrafish Traumatic Brain Injury Model in Assessing Cerebral Inflammation.

Traumatic brain injury (TBI) is a major public and socioeconomic problem throughout the world. The establishment of an effective and cost-effective TBI model for developing new therapeutic agents is challenging. Microglia are considered the resident macrophages of the central nervous system (CNS) that normally do not enter the brain. As the primary mediators of the innate immune response in the CNS, microglia play a critical role in neuroinflammation and secondary injury after TBI. In this study, we established an in vivo TBI zebrafish model using Tg(coro1a:EGFP) line where the green fluorescent protein-labeled microglia were present. We demonstrated that microglia accumulated rapidly in response to neuronal injuries. To clear away injured neurons and restore the CNS homeostasis, activated microglia secreted two types of functional cytokines, including pro-inflammatory interleukins (IL) of IL-1β and IL-6 and anti-inflammatory factors of IL-4 and IL-10 in the lesioned larvae. Cytidine 5'-Diphosphocholine (CDP-choline), as an effective and clinical neuroprotective drug, could further activate microglia, expressing high levels of il-1β, il-6, il-4, and il-10 in the TBI model. Moreover, CDP-choline reduced neuronal apoptosis and promoted neuronal proliferation around the lesioned site. Based on these results, the TBI model established in this study represents a suitable model for developing new therapeutic agents for CNS-associated diseases.

Rapid and Durable Responses with the SYK/JAK Inhibitor Cerdulatinib in a Phase 2 Study in Relapsed/Refractory Follicular Lymphoma-Alone or in Combination with Rituximab

Rapid and Durable Responses with the SYK/JAK Inhibitor Cerdulatinib in a Phase 2 Study in Relapsed/Refractory Follicular Lymphoma-Alone or in Combination with Rituximab

Cytokine Profile and Endothelial Status in Patients with Coronary Artery Disease and Different Outcomes of Coronary Artery Bypass Grafting

Aim.To assess the cytokine profile and the level of endothelial dysfunction in patients with coronary artery disease (CAD) and different outcomes of coronary artery bypass grafting (CABG).Materials and methods.The study included 95 people, with 22 of them being practically healthy (without CAD). The CAD patients (73) were retrospectively divided into 2 subgroups according to the presence of complications within the first month after the CABG. The first subgroup consisted of 25 patients suffering from complications after CABG, with 48 people making up the second subgroup (no complications after CABG). CABG was performed on a beating heart; bypass grafts were attached during revascularisation. The serum levels of IL-4 and IFN-γ were determined through the enzyme-linked immunosorbent assay (ELISA) using appropriate test systems manufactured by BCM Diagnostics and a Zenyth 340 microplate photometer (Biochrom Ltd). In order to determine the content of endothelin-1 in blood serum, ELISA test systems manufactured by Biomedica were used. The statistical analysis of study results was performed employing the descriptive statistics module of Statistica 12.0 (StatSoft, USA). The mean (M), its error (m), median (Me) and the interquartile range [Q25; Q75] were calculated. In cases when p ≤ 0.05, the results were considered statistically significant.Results.It was revealed that, in the preoperative period, CAD patients from both subgroups exhibited increased levels of endothelin-1 (p = 0.0001) and IFN-γ (p = 0.0001), whereas an increase in IL-4 production was noted only in the first subgroup (p = 0.0001), as compared to the control group. The follow-up revealed that CAD patients with complications after CABG exhibited stable high levels of IL-4 (p > 0.05) and IFN-γ (p > 0.05) during the entire observation period, as compared to the initial values. The content of endothelin-1 in both subgroups increased significantly on the 4th day of observation (p > 0.05) and returned to its initial high values after 1 month.Conclusion.Immune response in CAD patients suffering from complications after CABG develops mainly along the pro-inflammatory pathway and is accompanied by endothelial dysfunction.

COMPARISON OF PATHOGENESIS OF P. BERGHEI INFECTION IN MOUSE AND RAT MODELS

Background: The cytokine cascade in the immunopathogenesis of malaria infection had been widely studied. However, their specific association with survival and severe infection remained obscure. Methods: The study investigated the cytokine profiles and histopathological features of malaria in the severe infection and survival models by using male ICR mice and male Sprague Dawley rats respectively. Results: The severe model, the infected ICR mice, exhibited a high parasitemia with 100% mortality after peak parasitemia at day 5 post-infection. The survival model, the infected Sprague Dawley rats, showed mild parasitemia with full recovery by day 14 of infection. Both severe and survival models showed similar histopathological severity during peak parasitemia. The severe model produced highly elevated levels of proinflammatory cytokines, TNF-I± and IL-1I±, and low levels of the anti-inflammatory cytokine, IL-4; while the survival model showed low levels of TNF-I± and IL-1I± with high levels of IL-4. Conclusion: There were differences in the pathogenesis of the severe and survival models of malaria infection. These could be a basis for immunotherapy of malaria in the future.

Cocaine Use Disorder Is Associated With Changes in Th1/Th2/Th17 Cytokines and Lymphocytes Subsets.

Background: Cocaine is a psychostimulant drug with high addictive proprieties. Evidence suggests that cocaine use leads to critical changes in the immune system, with significant effects on T, B, and natural killer (NK) cells and influencing peripheral levels of cytokines. The presence of abstinence-related symptoms during detoxification treatment is known to influence the prognosis. Here, our aim was to investigate immune profiles in women with cocaine use disorder (CUD) according to withdrawal symptoms severity.Methods: Blood samples and clinical data were collected at onset of detoxification treatment of 50 women with CUD. The patients were stratified according to Cocaine Selective Severity Assessment (CSSA) scores in low withdrawal (L-W) and high withdrawal (H-W) categories. In addition, we also included a control group with 19 healthy women as reference to immune parameters. Peripheral blood was collected and lymphocyte subsets were phenotyped by multi-color flow cytometry (B cells, CD4+ T, CD8+ T, NK cells, and different stages of T-cell differentiation). PBMCs from patients and healthy controls were stimulated in vitro with phytohemagglutinin (1%) for 72 h to assess the production of Th1/Th2/Th17 cytokines.Results: Following stimulation, lymphocytes from women with CUD produced increased levels of Th1/Th2/Th17 cytokines. However, higher levels of IL-2 and IL-17 were observed only in the L-W group, while higher levels of IL-6 were detected in the H-W group compared to controls. H-W group showed lower percentage of early-differentiated Th cells (CD4+CD27+CD28+), elevated percentage of Th cells (CD3+CD4+), intermediate-differentiated Th cells (CD4+CD27−CD28+), and B cells (CD3−CD19+). Both CUD groups showed decreased percentages of naïve T cells (CD3+CD4+CD45RA+ and CD3+CD8+CD45RA+).Conclusion: Our data demonstrated that CUD can lead to increased production of Th1/Th2/Th17 cytokines and lymphocyte changes.

Features of the cytokine balance in infectious hemorrhagic colitis in children

The aim of the study was to study the features of cytokine balance and their effect on clinical and laboratory parameters in infectious hemorrhagic colitis in children. Materials and methods. In the Department of intestinal infections in 2018 observed 28 children with infectious haemorrhagic colitis. The diagnosis was established taking into account clinical recommendations; the severity of the disease was determined by the Clark index; the etiology was verified according to bacteriological and molecular studies of feces, serological methods. In the acute period of the disease, the concentration of cytokines in the blood serum — TNF-a, IL-4, IL-6, IL-10 was studied by enzyme immunoassay. Mann-Whitney method, Kruskal-Wallis dispersion analysis, Pearson correlation method we used to estimate the obtained data.Results. The age structure of patients was dominated by young children (53.6%). Mild form of the disease was diagnosed in 32.1% of patients, moderate — in 67.9%. A high level of IL-4 was observed against a relatively low level of TNF-a, IL-6 and IL-10. Significantly higher concentrations of TNF-a and IL-6 were found in children with moderate form of the disease. The maximum values of IL-6 were observed in young children, IL-10 — in infants. In the moderate form of the disease, negative correlations of IL-4 with the number of leukocytes (r = –0.46; p = 0.05) and platelets (r = –0.48; p < 0.05) in the hemogram noted. Early age patients were found to have correlations of TNF-a with the relative number of rod neutrophils (r = 0.62; p < 0.01). There was a significant correlation of IL-4 with the relative number of segmental neutrophils (r = 0.49; p < 0.05). The relationship of TNF-a with the severity of the disease (r = 0.42; p < 0.05) revealed. The data on the possible suppressive effect of TNF-a and IL-10 on the synthesis of urea and the relationship of IL-6 with reactive changes in the liver obtained.Conclusion. Studies of cytokine balance in infectious hemorrhagic colitis in children showed the severity of systemic inflammatory response with the activation of Th-2 immune response. The correlation of TNF-a with the severity of the disease was established, which may have diagnostic and prognostic value.

Association of peripheral cytokine levels with cerebral structural abnormalities in schizophrenia

A large body of evidence indicates that both the altered cytokines that mediate the immune-inflammatory process and abnormal gray matter are associated with schizophrenia. Whether peripheral cytokines are related to cerebral structural abnormality remains unclear. Therefore, we aimed to investigate the association of peripheral cytokine levels with gray matter abnormalities at the whole brain level in schizophrenia. Forty-four outpatients with schizophrenia and 44 controls were recruited. The serum levels of interleukin-1 beta (IL-1β), IL-2, IL-6, IL-8, interferon-gamma (IFN-γ), transforming growth factor-beta (TGF-β), and IL-10 were measured using a quantitative chemiluminescence assay. High-resolution T1 weighted images were acquired from all subjects and processed using FreeSurfer software to obtain the cortical thickness, surface area, and cortical and subcortical gray matter volumes. The cytokines and cerebral structures were compared between patients and controls using analysis of covariance (ANCOVA). The association between the cytokines and whole cerebral structures was performed using stepwise linear regression. Patients had higher levels of IL-2, IL-6, IL-8, and IL-10 than controls. In patients, the IL-6 level was significantly associated with the cortical thickness in the left pars opercularis, right pars triangularis, left superior temporal gyrus, and right middle temporal gyrus, which showed structural differences between the two groups. Altered cytokine levels may be associated with particular but not all cortical abnormalities in schizophrenia, especially IL-6, which was significantly associated with the abnormal cortical thickness of the bilateral Broca’s area and temporal gyrus, which provided neuroimaging evidence to support the relationship between peripheral cytokines and the cerebral cortex in schizophrenia.

Association of T Cell–Derived Inflammatory Cytokines With Acute Kidney Injury and Mortality After Cardiac Surgery

IntroductionAnimal models of renal ischemia-reperfusion injury (IRI) demonstrate that interferon (IFN)-γ producing T-helper (Th)-1 cells worsen acute kidney injury (AKI), whereas interleukin (IL)-4– and IL-13–producing Th2 cells lead to repair. We tested the association of these cytokines with AKI and mortality in patients who underwent cardiac surgery.MethodsIn 1444 participants of a multicenter, prospective, observational cohort, we measured 10 plasma biomarkers before and after cardiac surgery (IFN-γ, IL-4, IL-13, tumor necrosis factor [TNF]-α, IL-1β, IL-2, IL-6, IL-8, IL-10, and IL-12p70) and combined these biomarkers using principal component analysis (PCA). We also tested independent associations of Th1 (IFN-γ) and Th2 (IL-4 and IL-13) biomarkers with clinical outcomes of postoperative AKI and 1-year mortality.ResultsAKI occurred in 492 participants (34%), and 1-year mortality occurred in 81 participants (6%). Within 6 hours after surgery, IFN-γ, IL-4, and IL-13 increased 2.1-, 6.0-, and 4.6-fold, respectively, from their preoperative levels. Patients with higher levels of IFN-γ had higher odds of AKI (adjusted odds ratio per log change, 1.35 [1.13, 1.6]) and mortality (1.51 [1.17, 1.94]). Patients with higher levels of IL-4 and IL-13 also had higher odds of AKI (1.26 [1.09, 1.46] and 1.4 [1.16, 1.69], respectively) and mortality (1.46 [1.18, 1.82] and 1.71 [1.27, 2.31], respectively). Adding biomarkers to the clinical variables through use of PCA improved the area under the curve by 0.01 for AKI and 0.04 for mortality, resulting in final areas under the curve of 0.85 (0.83–0.87) and 0.76 (0.70–0.81), respectively.ConclusionBoth Th1 and Th2 cytokines increased immediately after cardiac surgery and were associated with AKI and 1-year mortality. Our findings indicate activation of both Th1 and Th2 pathways after cardiac surgery rather than predominance of either pathway.

232 Identifying the mechanisms underlying the negative feedback loop between tight junction proteins and cutaneous microbial dysbiosis in atopic dermatitis

Reduced skin barrier function and dysbiosis of the skin microbiota are hallmarks of atopic dermatitis (AD). Both, reduced cutaneous barrier and dominance of Staphylococcus aureus (S. aureus) are believed to represent entry points of inflammation in AD, but how they regulate each other is not known. Tight junctions (TJs) in the skin are close contacts between plasma membranes of neighboring cells and important for barrier functioning. TJs have also been indicated to be essential for skin barrier function in AD, yet little is known about their interplay with S. aureus. Here, we investigated how skin barrier impairment contributes to the development of AD, focusing on the microbial shift towards S. aureus dominance in a mouse model. We found that barrier impairment induced by repeated tape stripping of the back skin leads to a significant downregulation of several TJ proteins (e.g. claudin-1, occludin) detected by RT-PCR ex vivo. Furthermore, an upregulation of S. aureus-associated pro inflammatory cytokines (e.g. IL-1ß, TNF-α, IL-6) was detected. Analyses of the skin microbiome affirm this finding by pointing to a shift within the microbial homeostasis characterized by a predominance of staphylococci after tape stripping. We applied the FITC dermatitis model to induce AD-like inflammation. As a result, sensitization of the mice at tape stripped skin resulted in a significant increase in ear swelling upon challenge one week later compared to non-tape stripped mice, as well as a more pronounced Th2-dominated immune response associated by an increased CD3+CD4+ cell population and higher levels of IL-4 and IL-13. These findings establish a model of skin barrier disruption which can be applied to study molecular mechanisms of AD pathophysiology. They also indicate an important role of the downregulation of TJs contributing to a staphylococci-dominated dysbiosis of the skin and AD development. To conclude, this model and findings should be used to develop treatment strategies for patients suffer from this disease.

Chimeric hepatitis B virus core particles displaying Neisserial surface protein A confer protection against virulent Neisseria meningitidis serogroup B in BALB/c mice.

PurposeThe primary goal of the present study was to explore and evaluate the highly conserved Neisserial surface protein A (NspA) molecule, fused with truncated HBV virus-like particles (VLPs), as a candidate vaccine against the virulent Neisseria meningitidis serogroup B (NMB).MethodsNspA was inserted into the major immunodominant region of the truncated hepatitis B virus core protein (HBc; amino acids 1–144). The chimeric protein, HBc-N144-NspA, was expressed from a prokaryotic vector and generated HBc-like particles, as determined by transmission electron microscopy. Further, the chimeric protein and control proteins were used to immunize mice and the resulting immune responses evaluated by flow cytometry, enzyme-linked immunosorbent assay, and analysis of serum bactericidal activity (SBA) titer.ResultsEvaluation of the immunogenicity of the recombinant HBc-N144-NspA protein showed that it elicited the production of high levels of NspA-specific total IgG. The SBA titer of HBc-N144-NspA/F reached 1:16 2 weeks after the last immunization in BALB/c mice, when human serum complement was included in the vaccine. Immunization of HBc-N144-NspA, even without adjuvant, induced high levels of IL-4 and a high IgG1 to IgG2a ratio, confirming induction of an intense Th2 immune response. Levels of IL-17A increased rapidly in mice after the first immunization with HBc-N144-NspA, indicating the potential for this vaccine to induce a mucosal immune response. Meanwhile, the immunization of HBc-N144-NspA without adjuvant induced only mild inflammatory infiltration into the mouse muscle tissue.ConclusionThis study demonstrates that modification using HBc renders NspA a candidate vaccine, which can trigger protective immunity against NMB.

Combination of oxymatrine and diammonium glycyrrhizinate significantly mitigates mice allergic contact dermatitis induced by dinitrofluorobenzene.

This study investigated the safety and effect of oxymatrine (OMT) and/or diammonium glycyrrhizinate (DG) on allergic contact dermatitis (ACD) induced by 1-fluoro-2,4-dinitrofluorobenzene (DNFB) in ICR mice. Mice were topically smeared with vehicle (control) or DNFB on their ear and skin to induce ACD. The mice were randomized and injected with saline as the model, treated intraperitoneally with dexamethasone (DEX), 45 or 90 mg·kg−1 OMT and/or DG daily beginning one day post the first smearing for two weeks. The body weights, the severity of ear and skin inflammation, the levels of serum IgE, IL-4, and IFNγ, creatinine and urea as well as plasma sodium and potassium in individual mice were measured. In comparison with the control group, the model group did not change the body weights, but developed severe skin and ear inflammation with increased ear thickness, accompanied by many inflammatory infiltrates in the lesions and high levels of serum IgE, IL-4, and IFNγ. Combination of OMT and DG prevented the OMT- or DG-altered body weights in mice. While treatment with either OMT or DG moderately reduced the skin and ear inflammation, their thickness and inflammatory infiltrates, combination of OMT and DG further significantly increased their anti-inflammatory effects in mice. A similar pattern of inhibitory effect on the levels of serum IgE, IL-4, and IFNγ was observed in the different groups of mice. Combination of OMT and DG also prevented the OMT-, DG-, or DEX-altered plasma sodium or potassium levels in mice. Therefore, combination of OMT and DG significantly increased anti-inflammatory effects on ACD induced by DNFB in mice and attenuated DG- or OMT-related adverse effects. Impact statement Diammonium glycyrrhizinate (DG) and oxymatrine (OMT) have similar anti-inflammatory, anti-allergic, anti-tumor, immunomodulatory, and other pharmacological properties. Our previous study has shown that when DG and OMT are combined, DG can attenuate both high-dose (347.44 mg·kg−1) and regular-dose (90 mg·kg−1) OMT-induced mortality and adverse effects (such as body weight loss and hyponatremia). Furthermore, OMT can similarly attenuate the adverse effects (such as body weight gain, hypernatremia, and hypokalemia) induced by regular dose (90 mg·kg−1) of DG. Accordingly, we tested whether combination of OMT and DG would increase anti-inflammatory activities and reduce their adverse effect in a mouse model of allergic contact dermatitis (ACD) induced by 1-fluoro-2,4-dinitrofluorobenzene (DNFB). Our findings indicated that combination of OMT and DG significantly increased anti-inflammatory effects on ACD induced by DNFB in ICR mice and attenuated adverse effects of DG or OMT alone.

Abstract 5036: Induction of highly efficacious anti-tumor activity and modulation of tumor microenvironment: Cell-free off the shelf therapeutic modality

Abstract Adoptive transfer of Bispecific antibody Armed activated T cells (BATs) show promising anti-tumor activity in clinical trials in solid tumors. The cytotoxic activity of BATs occur upon engagement with tumor cells via the bispecific antibody bridge which stimulates BATs to release not only the lytic and cytotoxic molecules (perforin/granzyme) but also cytokines, chemokines and other signaling molecules extracellularly. We hypothesized that the release of extracellular soluble factors by this complex interaction of T cells, bispecific antibody, and tumor cells may serve as a potent anti-tumor and immune activating conditioned media (CM). In a 3D tumorsphere model, tumor+BATs-CM (n=10) showed potent cytotoxicity (p<0.001) against multiple breast (MDA-MB-231, BT-20, SK-BR-3 and MCF-7) and other cancer cell lines (p<0.001) compared to control tumor-CM or BATs-CM. Tumor+BATs-CM (n=6) was able to reduce the proportion of CD44hi/CD24lo cancer stem like cells to 0.7% compared to 4.9% in control CM. The addition of tumor+BATs-CM decreased the proportions of T regulatory cells (5% to 1.1%; p<0.02) and myeloid derived suppressor cells (3.8% to 1.2%; p<0.03), but increased activation and proliferation of effector T cells in 3D cultures compared to control CM (n=3). Size based CM factionation showed that most activity is retained in the <50kDa but >10kDa fraction. Multiplex analysis showed high levels of IL-2, IL-15, IFN-γ, TNF-α, GM-CSF, granszyme B (GZB), IL-13, MIP-1β, IP-10, MIG and RANTES. These factors are likely responsible for the cytolytic and immune activating effects. Phospho-specific signaling protein arrays showed enhanced JAK1/STAT-1/STAT-5A, Rac/cdc42/STIM-1) pathways in tumor+BATs-CM (n=3). Exosomal microRNA (miR) in tumor+BAT-CM showed higher expression of several miRs that are associated with T cell function and activation compared to control CM (n=2). Simulations using cocktails of multiple cytokines were done to test anti-tumor activity, IFN-γ/TNF-α/GZB showed potent cytotoxicity directed at breast (58-78%) and pancreatic cancer (50-72%) cell lines compared to 45-65%, 20-27%, 18-25% with IFN-γ, TNF-α and GZB individually, respectively. In a xenograft breast cancer model, IV and intra-tumoral injections of 10x concentrated tumor+BAT-CM (3x/week for 4 weeks;150μl CM/injection) was able to inhibit tumor growth significantly (p<0.01) compared to the control CM treated mice (n=10 mice/group). Therapeutic advantages of CM include: 1) a ready off-the-shelf product; 2) a decrease in regulatory and manufacturing costs. In summary, BATs-Tumor complex derived CM provides a clinically controllable cell-free platform to target various tumor types with diverse anti-cancer immune activating mediators regardless of the heterogeneous nature of the tumor cells and mutational burden as a novel and potent off-the-shelf therapeutic modality. Citation Format: Archana Thakur, SriVidya Yarlagadda, Kyungmin Ji, Dana L. Schalk, Johnson Ung, Edwin T. Bliemeister, Amro Aboukameel, Eli Casarez, Bonnie F. Sloane, Lawrence G. Lum. Induction of highly efficacious anti-tumor activity and modulation of tumor microenvironment: Cell-free off the shelf therapeutic modality [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 5036.

Serum leptin levels correlate negatively with the capacity of vitamin D to modulate the in vitro cytokines production by CD4+ T cells in asthmatic patients

Both obesity and low vitamin D levels have been associated with allergic asthma (AA) severity. In the present study, severity of AA was associated with obesity but to the in vitro IgE production. In those patients, higher levels of IL-5, IL-6 and IL-17 were quantified in CD4+ T-cell cultures as compared with patients with mild and moderate AA. In addition, the lowest IL-10 levels were detected in the cell cultures from patients with a worse prognosis. Interestingly, the occurrence of AA elevates the plasma levels of leptin, and this adipokine was positively correlated with the release of IL-5, IL-6 and IL-17, but inversely correlated with IL-10 production, by CD4+ T-cells from patients. In AA-derived CD4+ T-cell cultures, 1,25(OH)2D3 was less efficient at inhibiting IL-5, IL-6 and IL-17 production, and up regulating IL-10 release, as those from healthy subjects. Interestingly, the in vitro immunomodulatory effects of vitamin D were inversely correlated with serum leptin levels. In summary, our findings suggested that obesity, probably due to the overproduction of leptin, negatively impacts AA as it favors imbalance between Th2/Th17 and regulatory phenotypes. The deleterious effects of leptin may also be due to its ability to counter-regulate the immunosuppressive effects of vitamin D.

The cross-talk between infiltrating macrophages and inflamed or malignant colonic epithelium promotes overexpression of ST6GALNAC1 and epithelial MUC1 tumor form MUC1-sTn

Abstract Patients with inflammatory bowel disease (IBD) have an increased risk of developing colitis-associated colon cancer (CACC). Despite the strong relationship between inflammation and cancer, the mechanistic events that contribute to the transition from IBD to CACC remain undefined. Changes in the glycosylation profile of the known oncoprotein MUC1 commonly occur in chronic inflammation and may facilitate progression to cancer. We used a transwell coculture model system to examine the effect of polarized macrophages, such as those contributing to inflammation, on the expression of glycosylation-related enzymes in colon cancer cells. We found that M2-like macrophages induce the expression of ST6GALNAC1 glycosyltransferase which, by adding sialic acid to O-linked GalNAc residues, promotes the formation of the tumor-associated MUC1-sTn glycoform. Cytokine antibody arrays and blocking antibody experiments revealed that high levels of IL-13 and CCL17, present in the conditioned medium of colon cells cocultured with M2 macrophages, activate ST6GALNAC1 expression in colon cancer cells. In silico and in vivo murine and human models of colitis and CACC showed that IL-13 induces the phosphorylation of STAT-6 that directly binds ST6GALNAC1 promoter resulting in its transcription activation and protein over-expression. On the other hand, CCL17 activates ST6GALNAC1 expression via NF-kB pathway signaling. Our findings revealed a novel cross-talk between M2-like macrophages and inflamed and malignant colon cells that contributes to the pathogenesis of colitis and progression to CACC.

Circulating cytokines predict severity of rheumatic heart disease

BackgroundRheumatic heart disease (RHD) is associated with inflammation that damages cardiac valves, often requiring surgical interventions. The underlying mechanisms involved in the disease progression are not completely understood. This study aimed to evaluate cytokine plasma levels in patients with RHD as possible markers of disease severity. Methods and resultsEighty-nine patients with RHD, age of 41 years ±11.5 years, were prospectively enrolled. RHD severity was defined as valve dysfunction that required invasive intervention, either valve repair or replacement. Peripheral blood samples were collected from all patients for cytokine measurements. The patients were followed up to look at adverse clinical events defined as either the need for valve intervention or death.At baseline, 64 (71.9%) patients had previously undergone valve intervention, whereas 25 patients had stable clinical presentation. Patients with severe RHD displayed higher levels of inflammatory cytokines than patients with stable disease. Cluster analysis showed segregation of severe and stable RHD based on IL-6/TNF-α and IL-6/IL-17A, respectively. IL-6 and TNF-α expression were positively correlated in severe but not in stable RHD patients.During a median follow-up of 23 months, 16 patients (18%) had an adverse outcome. IL-10 at baseline (HR 1.24, 95% CI 1.08–1.43, p = 0.003), and IL-4 (HR 1.12, 95% CI 1.01–1.24, p = 0.041) were predictors of events during the follow-up. ConclusionsHigh levels of cytokines are associated with severity of RHD. The co-regulated expression of IL-6 and TNF-α is associated with severe valve dysfunction, whereas high IL-10 and IL-4 levels predicted subsequently adverse outcome.

Expression of TLR2, TLR3, TLR4, and TLR7 on pulmonary lymphocytes of Schistosoma japonicum-infected C57BL/6 mice

Despite the paramount role of TLRs in the induction of innate immune and inflammatory responses, there is a paucity of studies on the role of TLRs in Schistosoma japonicum infection. Here, we observed obvious infiltration of inflammatory cells in S. japonicum-infected C57BL/6 mouse lungs. Expression and release of IFN-γ, IL-4, and IL-17 were significantly higher in pulmonary lymphocytes from infected mice compared with control mice in response to anti-CD3 plus anti-CD28 mAbs. Higher percentages of TLR2, TLR3, TLR4, and TLR7 were expressed on such lymphocytes, and the TLR agonists PGN, Poly I:C, LPS, and R848 induced a higher level of IFN-γ. However, a higher level of IL-4 was found in the supernatant of pulmonary lymphocytes from infected mice stimulated by these TLR agonists plus CD3 Ab. Only R848 plus anti-CD3 mAb could induce a higher level of IFN-γ in such lymphocytes. TLR expressions were then compared on different pulmonary lymphocytes after infection, including T cells, B cells, NK cells, NKT cells, and γδT cells. The expression levels of TLR3 on T cells, B cells, NK cells, and γδT cells were increased in the lungs after infection. NK cells also expressed higher levels of TLR4 after infection of control mice. Collectively, these findings highlight the potential role of TLR expression in the context of S. japonicum infection.

480 The characteristics of patients with persistent HHV-6 infection after drug-induced hypersensitivity syndrome/drug reaction with eosinophilia and systemic symptoms (DIHS/DRESS)

Background: Drug-induced hypersensitivity syndrome/drug reaction with eosinophilia and systemic symptoms (DIHS/DRESS) is characterized by fever, cutaneous eruptions, hematological abnormalities, organ disorder, and reactivation of human herpesvirus 6 (HHV-6). In addition, autoimmune diseases such as type 1 diabetes and thyroiditis may develop long-term after resolution of DIHS. Reactivation of HHV-6 has been reported to be involved in clinical condition of DIHS. We found that quite a few patients harbor high levels (> 1000 copies / 106 PBMC) of HHV-6 DNA in PBMC over 4 months after onset of DIHS. However, it is unclear what effects persistent HHV-6 infection has on DIHS patients in chronic phase. Methods: We analyzed clinical symptoms, blood test findings, reactivation of herpes virus, expression of serum cytokines (IL-4, IL-5, IL-10, IFN-γ) and soluble IL-2 receptor for 8 DIHS patients with persistent HHV-6 infection and 10 patients with transient HHV-6 infection. Results: Compared to the transient HHV-6 infection group, persistent HHV-6 infection group showed 1) more severe acute phase cutaneous and mucosal eruptions, 2) higher levels of HHV-6 DNA in acute phase, 3) higher rate of cytomegalovirus reactivation, 4) higher levels of IL-4 in acute phase and higher levels of soluble IL-2 receptor in late phase, 5) higher rate of long-term complications (ie, interstitial nephritis, arthritis, thyroiditis). Conclusion: Persistent HHV-6 infection may have some influence on the pathological and immunological conditions of DIHS and may be involved in the clinical course and prognosis.

Group 2 Innate Lymphoid Cells Are Redundant in Experimental Renal Ischemia-Reperfusion Injury.

Acute kidney injury (AKI) can be fatal and is a well-defined risk factor for the development of chronic kidney disease. Group 2 innate lymphoid cells (ILC2s) are innate producers of type-2 cytokines and are critical regulators of homeostasis in peripheral organs. However, our knowledge of their function in the kidney is relatively limited. Recent evidence suggests that increasing ILC2 numbers by systemic administration of recombinant interleukin (IL)-25 or IL-33 protects against renal injury. Whilst ILC2s can be induced to protect against ischemic- or chemical-induced AKI, the impact of ILC2 deficiency or depletion on the severity of renal injury is unknown. Firstly, the phenotype and location of ILC2s in the kidney was assessed under homeostatic conditions. Kidney ILC2s constitutively expressed high levels of IL-5 and were located in close proximity to the renal vasculature. To test the functional role of ILC2s in the kidney, an experimental model of renal ischemia-reperfusion injury (IRI) was used and the severity of injury was assessed in wild-type, ILC2-reduced, ILC2-deficient, and ILC2-depleted mice. Surprisingly, there were no differences in histopathology, collagen deposition or mRNA expression of injury-associated (Lcn2), inflammatory (Cxcl1, Cxcl2, and Tnf) or extracellular matrix (Col1a1, Fn1) factors following IRI in the absence of ILC2s. These data suggest the absence of ILC2s does not alter the severity of renal injury, suggesting possible redundancy. Therefore, other mechanisms of type 2-mediated immune cell activation likely compensate in the absence of ILC2s. Hence, a loss of ILC2s is unlikely to increase susceptibility to, or severity of AKI.

Chlamydia muridarum infection induces CD4+ T cells apoptosis via PI3K/AKT signal pathway.

Apoptosis is essential for the homeostatic control of the lymphocytes number during the development of an immune response to an invasive microorganism. CD4+ T cells play a major role in homeostasis of the immune system and are sufficient to confer protection against Chlamydia muridarum (Cm) infection in mice. The present study demonstrated that phosphatidylinositol 3-kinase (PI3K) p110δ mRNA and phosphorylation of protein kinase B (p-AKT) level were significantly increased in lung cells and spleen cells at day 3 and day 7 post-infection, p-AKT level was inhibited when adding PI3K inhibitor LY294002. Moreover, Cm infection induced high levels of IL-2/IL-2Rα in CD4+ T cells, which may relate to PI3K/AKT signal pathway activation. We observed that Cm infection significantly induced apoptosis of CD4+ T cells. The related apoptosis proteins Bcl-2 and Mcl-1 uneven expression levels were induced in CD4+ T cells by Cm infection. These findings provided in vivo and in vitro evidence that Cm infection induces CD4+ T cells apoptosis possibly via PI3K/AKT signal pathway.