Th2 Chemokines Research Articles

The Th1 chemokine MIG in Graves' ophthalmopathy.

Chemokine (C-X-C motif) ligand (CXCL)9/monokine induced by interferon (IFN)-γ (MIG) and its receptor, chemokine (C-X-C motif) receptor (CXCR)3, are involved in the pathogenesis of Graves' ophthalmopathy (GO). In tissues, recruited Type 1 helper (Th1) lymphocytes could cause an increased production of IFN-γ and tumor necrosis factor (TNF)-α, which in turn stimulate MIG secretion from these cells; all this creates an amplification feedback loop, and perpetuates the autoimmune process. In particular, MIG is secreted by fibroblasts and preadipocytes under the influence of IFN-γ. The IFN-γ induced MIG secretion in vitro, in GO fibroblasts and preadipocytes can be modulate by peroxisome proliferator-activated receptors (PPAR)-γ and PPAR-α activators. High levels of MIG in peripheral liquids could be considered as a marker of an especially Th1 orientated immune response. The presence of circulating MIG is associated with the active phase of GO. An increased concentrations of MIG in patients with GO reflect, at least in part, the orbital inflammation activity. Since MIG concentrations are significantly reduced during corticosteroids and/or radiotherapy treatments, with respect to control group and basal values of patients with GO, MIG has been proposed as a marker in the therapy choice for patients with GO. More studies are needed to investigate interactions between chemokines and cytokines in the GO pathogenesis and to evaluate whether MIG is a novel therapeutic target in these autoimmune disorders.

The Th1 chemokine MIG in Graves' Disease: a narrative review of the literature.

Type-1 helper (Th1) dependent chemokines, such as monokine induced by interferon (IFN)-γ (MIG) seem to contribute to the Graves' disease (GD) pathogenesis. The thyrocytes secrete the chemokine (C-X-C motif) ligand (CXCL)10 under the IFN-γ influence. Therefore, high levels of MIG in peripheral liquids indicate a Th1 orientated immune response and are associated with the active phase of GD in hyperthyroid patients (newly diagnosed and relapsing). Methimazole (MMI), used for the hyperthyroidism treatment, causes a reduction of the MIG secretion by isolated thyrocytes, a decrease of serum MIG levels and leads to a shift from a Th1 to Th2 response in patients with GD in the active phase. The Th1 lymphocytes recruited in the tissues enhance the IFN-γ and tumor necrosis factor (TNF)-α production, that in turn stimulate MIG secretion from these cells; this mechanism originates an amplification feedback loop, causing a perpetuation of the autoimmune process. It has been seen that peroxisome proliferator-activated receptors (PPAR)-γ and PPAR-α activators can modulate the IFN-γ induced MIG secretion in vitro, in GD thyrocytes. More studies are needed to examine the interactions between cytokines and chemokines in the GD pathogenesis and to evaluate the role of MIG as a new therapeutic target.

Runx/Cbf\u03b2 complexes protect group 2 innate lymphoid cells from exhausted-like hyporesponsiveness during allergic airway inflammation

Group 2 innate lymphoid cells (ILC2s) have tissue-resident competence and contribute to the pathogenesis of allergic diseases. However, the mechanisms regulating prolonged ILC2-mediated TH2 cytokine production under chronic inflammatory conditions are unclear. Here we show that, at homeostasis, Runx deficiency induces excessive ILC2 activation due to overly active GATA-3 functions. By contrast, during allergic inflammation, the absence of Runx impairs the ability of ILC2s to proliferate and produce effector TH2 cytokines and chemokines. Instead, functional deletion of Runx induces the expression of exhaustion markers, such as IL-10 and TIGIT, on ILC2s. Finally, these ‘exhausted-like’ ILC2s are unable to induce type 2 immune responses to repeated allergen exposures. Thus, Runx confers competence for sustained ILC2 activity at the mucosa, and contributes to allergic pathogenesis.

Epigenetic modulation enhances immunotherapy for hepatocellular carcinoma

BackgroundAnti-PDL-1 immunotherapy for Hepatocellular Carcinoma (HCC) demonstrated a mixed response. Polycomb Repressor Complex 2(PRC2) contributes to the initiation and progression of HCC by suppressing tumor antigens and inhibiting an immune response. Two components of epigenetic modulation are Enhancer of Zeste Homolog 2 (EZH2, the catalytic component of PRC2) and DNA Methyltransferase 1 (DNMT1). We aim to investigate the potential role of epigenetic therapy targeting EZH2 and DNMT1 as a novel strategy to modulate immunotherapy response in HCC. MethodsHepG2, Hep3B, and Hepa1-6 HCC cell lines were treated with EZH2 inhibitor (DZNep) and DNMT1 inhibitor (5-Azacytidine) with and without anti-PDL-1. Quantitative RT-PCR and immunohistochemistry were performed to evaluate the expression of tumor suppressors, tumor antigens, and Th1 chemokines. In-vivo C57/LJ immunocompetent mice model with subcutaneous tumor inoculation was performed with intraperitoneal drug injections. ResultsThere was a significant upregulation of Th1 chemokines in HepG2 (CXCL9 5.5 ± 0.2 relative fold change; CXCL10 1.44 × 103 ± 37 relative fold change) and Hep3B (CXCL 9 6.85 × 103 ± 1.3 × 103 relative fold change; CXCL 10 2.15 × 103 ± 3.1 × 102 relative fold change). Additionally, there was a significant induction of cancer testis antigens NY-ESO-1 (3.6–3.7 ± 0.3 relative fold change) and LAGE (8.3–11.7 ± 1.9 relative fold change). In vivo model demonstrated statistically significant tumor regression in the combination treatment group (0.02 g ± 0.02) compared to epigenetic therapy (0.63 g ± 0.61) or immunotherapy alone (0.15 g ± 0.21) with untreated control (2.4 g ± 0.71). There was significantly increased trafficking of cytotoxic T- lymphocytes and associated apoptosis for the combination treatment group compared to epigenetic or immunotherapy alone. ConclusionsThis study demonstrates that epigenetic modulation could be a novel potential strategy to augment immunotherapy for HCC by stimulating T cell trafficking into tumor microenvironment via activation of transcriptionally repressed chemokine genes responsible for T-cell trafficking, inducing previously silent neoantigens for immune targets, and allowing tumor regression as a result. A clinical trial of this feasible combination therapy of these clinically available agents is warranted.

Role of Th 1- and Th 2- Chemokine Receptor in the Diagnosis and Prognosis of Primary Immune Thrombocytopenia.

The diagnostic and prognostic role of Th-1 chemokine receptor and Th-2 chemokine receptor in patients with primary immune thrombocytopenia has not been investigated extensively so far. In this study, our goal is to explore the diagnostic and prognostic role of C-C chemokine receptor 3 (CCR3) and C-C chemokine receptor 5 (CCR5) in patients with primary immune thrombocytopenia. The expression levels of CCR3 and CCR5 were measured in peripheral blood mononuclear cells of pa-tients with primary immune thrombocytopenia and healthy subjects. The relationship between the expression levels of CCR3 and CCR5 and clinicopathological characteristics was analyzed. The diagnostic accuracy of CCR3 and CCR5 as biomarkers to discriminate primary immune thrombocytopenia patients from healthy subjects was determined. Univariate and multivariate Cox regression analysis were performed to determine the prognosis value of CCR3 and CCR5 in primary immune thrombocytopenia. The outcome of primary immune thrombocytopenia patients was also evaluated. Compared to healthy subjects, the expression level of CCR3 was significantly downregulated and CCR5 was significantly upregulated (p < 0.05). The expression levels of CCR3 and CCR5 were significantly correlated with bleeding times and platelet counts at diagnosis (p < 0.05). CCR3 and CCR5 could act as a suitable biomarker for differentiating the primary immune thrombocytopenia patients from healthy subjects. CCR3 and CCR5 were independent prognostic factors. Overexpression of CCR5 and low expression of CCR3 lead to poor clinical benefits and indicated poor prognosis of primary immune thrombocytopenia. To summarize, our results suggested that CCR3 and CCR5 could act as suitable biomarkers and indicated poor prognosis of primary immune thrombocytopenia.

Characterization of a Mouse Model of Hypereosinophilic Syndrome

Introduction: Hypereosinophilic syndrome is characterized by sustained blood and/or tissue hypereosinophilia leading to tissue damage and organ dysfunction. Nearly half the cases are classified as idiopathic, since no known cause is identified and there is no evidence of a clonal disorder. In such cases, nonspecific treatment approaches, including corticosteroids, hydroxyurea and interferon-alpha, are used aimed at mitigating eosinophil-mediated organ damage. However, mortality occurs primarily due to cardiac and thromboembolic complications. Understanding the cause and mechanism of disease would aid in the development of targeted therapies with greater efficacy and fewer side effects. For instance, while the traditional view is that eosinophils are detrimental via toxicity of their granule proteins, recent studies suggest other mechanisms including non-canonical cell death subroutines, such as regulated necrosis, release of pro-thrombotic DNA “traps” and others. Lack of a mouse model hinders testing such hypotheses in vivo. We have developed a mouse model of eosinophil-associated hypereosinophilic syndrome characterized by multi-organ involvement including lethal prothrombotic myocardial damage.Results: We discovered a spontaneous mouse mutant in our colony with a hypereosinophilic phenotype. Mice develop peripheral blood eosinophilia, infiltration of lungs, spleen and heart by eosinophils, and extensive myocardial damage and remodeling. This ultimately leads to heart failure and premature death, often by 14 weeks of age. Histopathologic assessment of the hearts from affected mice (Fig. 1) revealed a robust inflammatory infiltrate of the ventricular myocardium. Immunohistochemistry demonstrated that this infiltrate is composed primarily of eosinophils (MBP) and B lymphocytes (B220), leading to myocardial damage and replacement fibrosis. In some cases, hearts showed dilatation and thinning of the right ventricular wall, suggestive of an inflammatory dilated cardiomyopathy. An exam of most mice revealed primarily atrial thrombi, which often filled the entire/majority of the chamber. Expression analysis by protein multiplex assay in heart homogenates revealed overexpression of Th1 and Th2 cytokines and chemokines, including IL-4, IL-6, LIF, IL-16, IP-10/Cxcl10, MIG/Cxcl9, Eotaxin-1/Ccl11, RANTES/Ccl5, MIP-1β/Ccl4, MIP-3β/Ccl19, MCP-1/Ccl2, MCP-3/Ccl7, MDC/Ccl22 and TARC/Ccl17.Conclusion: The pathologies observed in the mutant line are reminiscent of those seen in patients with hypereosinophilia, where cardiac-related morbidities, like congestive heart failure and thrombi, are the most common causes of death. As such, our model provides an opportunity to test mechanistic hypotheses and develop targeted therapies. [Display omitted] DisclosuresNo relevant conflicts of interest to declare.

The Mantle Cell Lymphoma Response to VCR Protocol Associated with Its Sensitivity to Hypomethylation Regulation-a Collateral Study on VCR Phase I Trial

Introduction:Mantle cell lymphoma (MCL) is a malignancy of monomorphic small to medium-sized B lymphocytes and is considered incurable with standard chemotherapy. Median survival estimates are three to seven years, with even shorter survival times in patients with the blastoid variant and with higher proliferation rates. A number of chemotherapy regimens, both conservative and aggressive, have been employed to treat MCL. Many of these treatments, however, are highly toxic especially in elderly patients or patients with comorbidities. The lack of curative ability and high toxicity of current treatments gives rise to a need for a new regimen. Following a phase I trial utilizing bortezomib, cladribine and rituximab (VCR) (NCT01439750, Blood 2016 128:1792), a series of exploratory assays was utilized to help understand the effectiveness of this combination. It is well documented that cladribine has epigenetic effects and recently, it has been shown that epigenetic modulators play an important role in a variety of cancerous phenotypes. Considering this, we have hypothesized that a positive response to the VCR regimens can be linked to successful hypomethylation regulation of specific genes.Methods:Patients with relapsed or refractory MCL who met specific criteria were eligible to participate in this study. It was a single-arm open-label, investigator initiated phase 1 clinical trial to assess the safety and efficacy of combination treatment with bortezomib, cladribine, and rituximab. Following the completion of the clinical trial, a DNA methylation assay was performed to assess the biological factors involved in treatment response. Additionally, a cytokine analysis using a magnetic luminex assay was performed to further investigate possible biological dissimilarities that could account for differences in treatment response. The specific genes targeted in the cytokine analysis were PD-L1, CXCL12, IFN-gamma, Il-2, CXCL10, CXCL9, IL-15 and TNF.Results:A principal components analysis of the methylation data revealed that there were significant overall differences in the methylation patterns between patients who responded well and those who responded poorly to treatment. When evaluating genomic regions that differed between groups, it appeared that there was no difference in percentage of hypomethylated regions vs. hypermethylated regions. An analysis of specific genes that have known links to either cancer growth or B-cell production and growth, however, revealed possible targets to show epigenetic differences. Cytokine levels as well as methylation status at the TNF, PD-L1 and CXCL-12 genes were different between the two groups. Those who responded well exhibited significantly decreased methylation percentage at these genes whereas those who responded poorly indicated increased methylation or no change at all.Discussion:Using the methylation and cytokine assay results, we attempted to understand both the positive and negative outcomes of the drug regimen via the patient's genomic response. A previous study done with an ovarian cancer model showed that the TH1 chemokines CXCL-9 and CXCL-10 play a large role in epigenetic tumor modulation [Peng, et al, Nature. 2015, 527 (7577):249-53]. Despite not seeing any significant differences in these particular genes, the methylation changes noted at the CXCL-12 locus indicate a possible mechanism of detecting treatment effectiveness. The CXCR4/CXCL-12 axis plays an important role in cancer cell growth and treatment evasion. Activity in this area shows that the tumor is adapting to treatment, indicating effectiveness as well as a need for further intervention. Changes in the promoter region of PD-L1 and TNF are also possible targets for biomarkers of treatment effectiveness. DisclosuresNo relevant conflicts of interest to declare.

Immune response to purified protein derivative in infants from helminth-sensitized mothers - A cases series.

Maternal exposure to antibodies, cytokines or parasitic antigens during gestation may alter the degree of immune competence of offspring. Here we describe the production of cytokines and chemokines, and the ability to activation of the immune response in infants from mothers sensitized to helminths. It were selected five infants born to helminth-seropositive mothers but who were negative for current helminth infection. Whole blood was cultured without stimulus, with phytohemagglutinin mitogen (5 μg/ml, 24 h) or with purified protein derivative (PPD) (1 μg/ml, 24 h), and the supernatant was assessed for presence of Th1/Th2 cytokines (IFN-γ, TNF-α, IL-10, IL-5, IL-4 and IL-2) and chemokines (CXCL10, CCL2, CXCL9, CCL5 and CXCL8) by cytometric bead array. All infants produced CCL5. Two infants demonstrated a mixed profile of Th1 (CXCL9) and Th2 (CCL2) chemokines in the presence of CXCL10, while one infant showed skewing towards Th2 without CXCL10 and two of them had been impaired immune response (children from sensitized to Schistosoma mansoni mothers). Infants with Th1 and Th2 profile chemokines demonstrated a good response to vaccination, indicated by CXCL10 levels, but not infants predominantly Th2-skewed profile. These results highlight that children from mothers sensitized to S. mansoni may lead to ineffective immune response to PPD, while mothers sensitized to Ascaris lumbricoides showed no such impairment.

Dietary ω-3 fatty acids alter the lipid mediator profile and alleviate allergic conjunctivitis without modulating Th2 immune responses.

Allergic conjunctivitis (AC) is one of the most common ocular surface diseases in the world. In AC, T helper type 2 (Th2) immune responses play central roles in orchestrating inflammatory responses. However, the roles of lipid mediators in the onset and progression of AC remain to be fully explored. Although previous reports have shown the beneficial effects of supplementation of ω-3 fatty acids in asthma or atopic dermatitis, the underlying molecular mechanisms are poorly understood. In this study, a diet rich in ω-3 fatty acids alleviated AC symptoms in both early and late phases without affecting Th2 immune responses, but rather by altering the lipid mediator profiles. The ω-3 fatty acids completely suppressed scratching behavior toward the eyes, an allergic reaction provoked by itch. Although total serum IgE levels and the expression levels of Th2 cytokines and chemokines in the conjunctiva were not altered by ω-3 fatty acids, eosinophil infiltration into the conjunctiva was dramatically suppressed. The levels of ω-6–derived proinflammatory lipid mediators, including those with chemoattractant properties for eosinophils, were markedly reduced in the conjunctivae of ω-3 diet–fed mice. Dietary ω-3 fatty acids can alleviate a variety of symptoms of AC by altering the lipid mediator profile.—Hirakata, T., Lee, H.-C., Ohba, M., Saeki, K., Okuno, T., Murakami, A., Matsuda, A., Yokomizo, T. Dietary ω-3 fatty acids alter the lipid mediator profile and alleviate allergic conjunctivitis without modulating Th2 immune responses.

Increased serum Th2 chemokine levels are associated with bronchopulmonary dysplasia in premature infants.

Bronchopulmonary dysplasia (BPD) is one of the most common chronic inflammatory lung disease of premature infants, with serious short- and long-term consequences. Early identification of premature infants at risk of BPD is critical to preventing the pathogenesis of disease. Thus, in the present study, we recruited 126 premature infants, collected peripheral blood samples at different time points during early life, and measured the concentration of Th1 (MCP-1, IP-10, and MIG) and Th2 (eotaxin-1, eotaxin-2, and MCP-4) chemokines in serum. We found serum eotaxin-2 levels were significantly higher in the BPD group than in the non-BPD group on day 1 [1662pg/ml vs. 1221pg/ml, P< 0.05], day 7 [1533pg/ml vs. 1089pg/ml, P< 0.05], and day 14 [1246pg/ml vs. 704pg/ml, P< 0.05] after birth, and serum MCP-4 levels were also significantly higher in the BPD group than in the non-BPD group on day 1 [186pg/ml vs. 128pg/ml, P< 0.05], day 7 [199pg/ml vs. 101pg/ml, P< 0.05], and day 14 [238pg/ml vs. 106pg/ml, P< 0.05] of life.Conclusions: Increased levels of Th2 chemokines, eotaxin-2, and MCP-4, are associated with BPD in premature infants. What is Known: • The pathogenesis of BPD is multifactorial and it is difficult to predict and prevent. • Previous studies have demonstrated that inflammation plays a major role in the pathogenesis of BPD. What is New: • Increased Th2 chemokines, eotaxin-2 and MCP-4, were associated with BPD in premature infants. • Abnormal Th1/Th2 response in early life maybe associated with the subsequent development of BPD, which provide a new insight to understand the pathogenesis of the disease.

Minocycline decreases Th2 chemokines from M2 macrophages: Possible mechanisms for the suppression of bullous pemphigoid by traditional bullous disease drugs.

Minocycline/tetracycline is clinically used for the treatment of bullous pemphigoid (BP), and its clinical benefits are superior to those of prednisolone when considering adverse events. Although the clinical benefits of minocycline/tetracycline are well known, its immunosuppressive mechanisms are still unclear. In this study, we investigated the immunomodulatory effects of traditional anti-BP drugs (minocycline, nicotinic acid amide, dexamethasone and cyclosporine) on CD163+ M2 macrophages in vitro, with special focus on the production of CCL18 and CCL22, both of which are produced by CD163+ M2 macrophages in the lesional skin of BP and are increased in the serum of BP patients. Minocycline decreased the production of CCL22, CCL24 and CCL26 as well as CCL2 from M2 macrophages. CCL18 from M2 macrophages was decreased by dexamethasone and cyclosporine, but not decreased by minocycline. These data suggest that the clinical benefit of minocycline is partially explained by its suppressive effects against the production of specific Th2 chemokines from M2 macrophages, which should contribute to the recruitment of Th2 cells and eosinophils in the lesional skin of BP patients.

LB1595 SENP1 regulates TH1 chemokines via IFN-γ−STAT1 signaling in keratinocytes through STAT3-deSUMOylation negative feedback loop

LB1595 SENP1 regulates TH1 chemokines via IFN-γ−STAT1 signaling in keratinocytes through STAT3-deSUMOylation negative feedback loop

Flightless I Alters the Inflammatory Response and Autoantibody Profile in an OVA-Induced Atopic Dermatitis Skin-Like Disease.

Atopic dermatitis (AD) is a chronic pruritic inflammatory skin disease characterized by excessive inflammation and disrupted skin barrier function. Although the etiology of AD is not completely understood, clinical and basic studies suggest increasing involvement of autoantibodies against intracellular proteins. An actin remodeling protein, Flightless I (Flii), has been shown to promote development of inflammatory mediated skin conditions and impairment of skin barrier development and function. Here, we sought to determine the effect of altering Flii expression on the development of AD and its contribution to autoimmune aspects of inflammatory skin conditions. Ovalbumin (OVA)-induced AD skin-like disease was induced in Flii heterozygous (Flii+/−), wild-type (Flii+/+), and Flii transgenic (FliiTg/Tg) mice by epicutaneous exposure to OVA for 3 weeks; each week was separated by 2-week resting period. Reduced Flii expression resulted in decreased disease severity and tissue inflammation as determined by histology, lymphocytic, and mast cell infiltrate and increased anti-inflammatory IL-10 cytokine levels and a marked IFN-γ Th1 response. In contrast, Flii over-expression lead to a Th2 skewed response characterized by increased pro-inflammatory TNF-α cytokine production, Th2 chemokine levels, and Th2 cell numbers. Sera from OVA-induced AD skin-like disease Flii+/− mice showed a decreased level of autoreactivity while sera from FliiTg/Tg mice counterparts showed an altered autoantibody profile with strong nuclear localization favoring development of a more severe disease. These findings demonstrate autoimmune responses in this model of OVA-induced AD-like skin disease and suggest that Flii is a novel target, whose manipulation could be a potential approach for the treatment of patients with AD.

NK-4 exerts selective regulatory effects on the activation and function of allergy-related Th2 cells.

NK-4 is the main component of the antiallergic drug Lumin, which has been in popular usage since the early 1950s. In this study, we examined whether NK-4 exerts a regulatory effect on the activation and effector function of Th2 cells. NK-4 inhibited IL-4 production by anti-CD3ε mAb-stimulated BALB/c mouse spleen cells, whereas NK-4 had little effect on IFN-γ production. IL-4 and IL-5 secretion by anti-CD3ε mAb- or antigen-stimulated Th2 cells (D10.G4.1) was abrogated by NK-4 without affecting cell numbers, whereas IFN-γ secretion by activated Th1 cells was unchanged. Mechanistic analysis revealed that NK-4 inhibited mRNA expression of the Th2-associated transcription factors GATA-3 and NFATc1 in anti-CD3ε mAb-stimulated D10.G4.1 cells. Regarding the regulation of Th2 cell effector functions, NK-4 inhibited the secretion of eotaxin and thymus and activation-regulated chemokine (TARC) by normal human dermal fibroblasts in response to IL-4 and/or TNF-α. NK-4 achieved TARC attenuation comparable to what is observed with suplatast tosilate, an antiallergic drug that selectively inhibits Th2 cytokine production, at 14-fold lower concentrations of suplatast tosilate. Dexamethasone increased TARC production by 2.2- to 2.6-fold of control cultures. NK-4 successfully inhibited the STAT6 signaling pathway, suggesting a potential mechanism for down-regulating chemokines expression. In addition, NK-4 abrogated IL-4-driven modulation of cytokine production profile in human monocytic THP-1 cells from proinflammatory to anti-inflammatory response, as seen in the inverted ratio of TNF-α to IL-10 produced in response to LPS. These results suggest that NK-4 could prevent IL-4-driven polarization to alternatively activated macrophages, which are proposed to have pathogenic roles in allergic asthma. The importance of Th2 cytokines and chemokines in the development and progression of type 2 inflammatory disorders has been highlighted by recent advance in our understanding the immunological mechanism underlying allergic disease. Our results support the use of NK-4 as a reasonable therapeutic option to alleviate Th2-mediated allergic inflammation.

SH2B3 (lnk) Regulates Type 1 Diabetes Immune Phenotypes

Background: The type 1 diabetes (T1D)-associated risk gene SH2B Adaptor Protein 3 (SH2B3) encodes a phosphatase with specificity for Janus Kinase 2 (JAK2). We hypothesized that variations in SH2B3 would affect immune signaling pathways relevant to T1D autoimmunity. Here we report: 1) Occurrence of the SH2B3 risk variant among T1D patients, unaffected first-degree relatives and controls; 2) Prevalence of SH2B3 risk genotype in African Americans in the Southeastern U.S.; and 3) Alterations to immune composition associated with the SH2B3 risk variant. Approach and Results: 1) The University of Florida Diabetes Institute (UFDI) cohort (n=1946) was analyzed for genetic risk of T1D using a Taqman single nucleotide polymorphism (SNP) genotyping array with coverage of 32 SNPs. Two SNPs (rs3184504 and rs653178) corresponding to SH2B3 were in perfect linkage disequilibrium and will be described here as minor (T1D risk) or common variant. Among Caucasian subjects, the odds ratio (OR) for the minor variant among T1D subjects was 1.30 (1.06-1.59, p=1.42x10-2), similar to previous reports. Among African American subjects, the minor variant OR was elevated (2.93, 1.22-7.03, p=1.30x10-2), representing the most highly-associated non-HLA gene. Complete blood count (CBC, n=954) and flow cytometric human immunophenotyping (HIP, n=252) were performed on a subset of subjects. Individuals carrying the risk variant of SH2B3 had significantly more blood monocytes and neutrophils but similar numbers of lymphocytes compared to non-carriers. HIP revealed a significantly higher frequency of type 1 (CXCR3+) CD8+ T cells in subjects with the risk variant of SH2B3. Conclusions: The risk variant of SH2B3 is associated with both increased risk for T1D and seroconversion in Caucasian and African American subjects. Phenotypically, this element of T1D genetic risk presents as increased frequency of some myeloid cell populations and either expansion or polarization of CD8+ T cells bearing the canonical Th1 chemokine receptor CXCR3. Disclosure A. Camargo: None. D.J. Perry: None. T.M. Brusko: Stock/Shareholder; Self; OneVax, LLC. Advisory Panel; Self; Caladrius Biosciences, Inc.. Consultant; Self; Merck &amp; Co., Inc., Sanofi-Aventis. M.A. Wallet: None.

Protective Effects of Casticin From Vitex trifolia Alleviate Eosinophilic Airway Inflammation and Oxidative Stress in a Murine Asthma Model.

Casticin has been isolated from Vitex trifolia and found to have anti-inflammatory and anti-tumor properties. We also previously discovered that casticin can reduce pro-inflammatory cytokines and ICAM-1 expression in inflammatory pulmonary epithelial cells. In the present study, we evaluated whether casticin reduced airway hyper-responsiveness (AHR), airway inflammation, and oxidative stress in the lungs of a murine asthma model and alleviated inflammatory and oxidative responses in tracheal epithelial cells. Female BALB/c mice were randomly divided into five groups: normal controls, ovalbumin (OVA)-induced asthma, and OVA-induced asthma treated with intraperitoneal injection of casticin (5 or 10 mg/kg) or prednisolone (5 mg/kg). Casticin reduced AHR, goblet cell hyperplasia, and oxidative responses in the lungs of mice with asthma. Mechanistic studies revealed that casticin attenuated the levels of Th2 cytokine in bronchoalveolar lavage fluids and regulated the expression of Th2 cytokine and chemokine genes in the lung. Casticin also significantly regulated oxidative stress and reduced inflammation in the lungs of mice with asthma. Consequently, inflammatory tracheal epithelial BEAS-2B cells treated with casticin had significantly suppressed levels of pro-inflammatory cytokines and eotaxin, and reduced THP-1 monocyte cell adherence to BEAS-2B cells via suppressed ICAM-1 expression. Thus, casticin is a powerful immunomodulator, ameliorating pathological changes by suppressing Th2 cytokine expression in mice with asthma.

CCL2 is Modulated by Cytokines and PPAR-γ in Anaplastic Thyroid Cancer.

Chemokine (C-C motif) ligand (CCL)2, the prototype Th2 chemokine, is secreted by tumor cells, and has growth promoting effects. Whether CCL2 protumorigenic activities will be validated, then CCL2 and its receptor CCR2 may be therapeutic targets in cancer. We tested in "primary human anaplastic thyroid carcinoma (ATC) cells" (ANA) versus "normal thyroid follicular cells" (TFC): a) CCL2 secretion basally, after IFN-γ and/or TNF-α stimulation; b) PPARγ activation by thiazolidinediones (TZDs), rosiglitazone or pioglitazone, on CCL2 secretion, and on proliferation and apoptosis in ANA. ANA produced basally CCL2, at a higher level versus TFC. IFN-γ or TNF-α dose-dependently induced the CCL2 release in 3/6 or 5/6 ANA, respectively, but in all TFC. IFN-γ+TNF-α induced a synergistic release of CCL2 in all TFC, but only in 1/6 ATC. TZDs exerted an inhibition of CCL2 release in 3/6 ANA, while had no effect in TFC. Pioglitazone inhibition of ANA proliferation was not associated with the effect on CCL2; NF-κB and ERK1/2 were basally activated in ANA, increased by IFN-γ+TNF-α, and pioglitazone inhibited IFN- γ+TNF-α activation. CCL2 serum levels were higher in 6 ATC patients than in 5 controls (813±345 versus 345±212, pg/mL; respectively; P<0.01, ANOVA). ANA produce CCL2 basally and after cytokines stimulation, with an extremely variable pattern of modulation, suggesting different types of deregulation in the chemokine modulation. Serum CCL2 is increased in ATC patients. Further studies will be necessary to evaluate if CCL2 might be used as a marker in the followup of ATC patients.

Exploring Immune Development in Infants With Moderate to Severe Atopic Dermatitis.

Atopic dermatitis (AD) is the most common chronic inflammatory skin disease in infancy with a complex pathology. In adults, the clinical severity of AD has been associated with increases in T helper cell type (Th) 2, Th22, and Th17 serum markers, including high levels of CC chemokine ligand (CCL) 17 and CCL22 chemokines. To explore the possible association between serum chemokine levels and AD severity in infants with moderate-to-severe AD and elevated immunoglobulin E (IgE). Serum samples (n = 41) obtained from a randomized, double-blind, and clinical dietary intervention study were used to study biomarkers in infants with AD. Baseline- and post-intervention samples (4 months) were used, six chemokines and nine ratios thereof were analyzed using Luminex and correlated to AD severity. In the initial study, the infants were randomized to receive extensively hydrolyzed whey-based formula without (control) or with short-chain galacto-oligosaccharides/long-chain fructo-oligosaccharides (9:1) and Bifidobacterium breve M-16V (active). 31 Infants up to 11 months of age, with an objective-SCORAD score (oSCORAD) ≥ 20 and elevated total-IgE and/or specific-IgE levels were included. In time, the median oSCORAD decreased in both groups by -8 (control, p < 0.05; active, p < 0.01). Irrespective of dietary intervention, several changes in Th2 chemokines (CCL17 and CCL22), inflammatory chemokine (CCL20), and the Th1 chemokine, CXC chemokine ligand (CXCL) 9, were detected over time. Overall CCL17 correlated to oSCORAD (r = 0.446, p < 0.01). After 4 months of dietary intervention, CXCL9 was higher (p < 0.01) in the active group compared with control [active, 2.33 (1.99-2.89); controls, 1.95 (1.77-2.43) log 10 median (range)]. In addition, a reduction in Th2/Th1 chemokine ratios for CCL17/CXCL9, CCL22/CXCL9, CCL20/CXCL10, and CCL20/CXCL11 was detected associated with the active intervention. While this study is small and exploratory in nature, these data contribute to immune biomarker profiling and understanding of AD in infants.

Early-life exposure to three size-fractionated ultrafine and fine atmospheric particulates in Beijing exacerbates asthma development in mature mice

BackgroundEpidemiological studies have suggested that elevated levels of air pollution contribute to an increased incidence or severity of asthma. Although late-onset adult asthma seems to be more attributable to environmental risk factors, limited data is available on the impact of early-life exposure to size-fractionated ambient particulate matter (PM) on asthma in adults. We aimed to determine the effect on the development and exacerbation of asthma in the adult after the mice were exposed as juveniles to three size-fractionated ambient particulates collected from Beijing.MethodsThe three size-fractionated ambient particulates were collected from urban Beijing in winter, heavily affected by traffic and coal-fired emissions. The typical morphological and major chemical components of the PM were characterized first. Oxidative stress and expression of DNA methyltransferases (DNMTs) were then examined in vitro and in the lungs of mouse pups 48 h after exposure to PM by oropharyngeal aspiration. When the exposed and control juvenile mice matured to adulthood, an antigen-induced asthma model was established and relevant bio-indices were assessed.ResultsPM with different granularities can induce oxidative stress; in particular, F1, with the smallest size (< 0.49 μm), decreased the mRNA expression of DNMTs in vitro and in vivo the most significantly. In an asthma model of adult mice, previous exposure as juveniles to size-fractionated PM caused increased peribronchiolar inflammation, increased airway mucus secretion, and increased production of Th2 cytokines and chemokines. In general, F1 and F2 (aerodynamic diameter < 0.95 μm) particulates affected murine adult asthma development more seriously than F3 (0.95–1.5 μm). Moreover, F1 led to airway inflammation in the form of both increased neutrophils and eosinophils in BALF. The activation of the TGF-β1/Smad2 and Smad3/Stat3 signaling pathways leading to airway fibrosis was more profoundly induced by F1.ConclusionThis study demonstrated that exposure to ambient PM in juvenile mice enhanced adult asthma development, as shown by increased Th2 responses, which might be associated with the persistent effects resulting from the oxidative stress and decreased gene expression of DNMTs induced by PM exposure. The observed differences between the effects of three size-fractionated particulates were attributed to particle sizes and chemical constituents, including heavy metals and also PAHs, since the amounts of PAH associated with more severe toxicity were enriched equivalently in the F1 and F2 fractions. Relative to the often mentioned PM2.5, PM with an aerodynamic diameter smaller than 0.95 μm had a more aggravating effect on asthma development.

Induction of Th1 chemokine secretion in dermal fibroblasts by vanadium pentoxide.

Vanadium is a soft, silvery‑grey metal with a number of different oxidation states. The most common commercial form of vanadium is vanadium pentoxide (V2O5). All vanadium compounds are considered toxic. An increase in skin rashes has been observed in certain vanadium workers, including the development of atopic dermatitis. However, to the best of our knowledge, no prior invivo or invitro studies have evaluated the effect of vanadium exposure in human dermal fibroblasts. The present study evaluated the effect of V2O5 on proliferation and chemokine secretion in dermal fibroblasts. The results revealed that V2O5 had no significant effect on the viability or proliferation of fibroblasts, however it was able to induce the secretion of T‑helper (Th)1 chemokines from dermal fibroblasts, synergistically increasing the effect of important Th1 cytokines, including interferon‑γ and tumor necrosis factor‑α. Through these processes, V2O5 may lead to the induction and perpetuation of an inflammatory reaction in dermal tissue. The induction and perpetuation of inflammation in the dermis and the variety of involved candidate genes may be at the base of V2O5‑induced effects following occupational and environmental exposures. Further studies are necessary to evaluate dermal integrity and manifestations in subjects who are occupationally exposed, or living in polluted areas.