Serum Levels Of CCL2 Research Articles

Atheroprotective Roles of Adiponectin via CCL2 Inhibition.

Aim: Adiponectin (APN) exhibits different atheroprotective effects, and we have previously reported that APN function is modulated by its binding proteins, E-selectin ligand 1, Mac-2 binding protein, and cystatin C. In the present study, we aimed to identify a novel atheroprotective mechanism of APN via C–C motif chemokine 2 (CCL2). Methods: We conducted iMAP ® -intravascular ultrasound (IVUS) in 111 Japanese male patients with stable angina. The plaque characteristics were determined where “plaque burden” [(EEM CSA − lumen CSA)/(EEM CSA)×100 (%)] ＞50%, and their correlation with serum CCL2 and APN levels was analyzed. Using western blot analysis, the effects of APN on the biological effects of CCL2 were examined in their mutual binding by co-immunoprecipitation assay, the monocyte migration, and the phosphorylation of MAP kinases. Results: In a clinical study, we found that the percentage of plaque in the culprit lesion was correlated positively with serum CCL2 and negatively with serum APN levels, with significance. We identified CCL2 as a novel APN-binding serum protein using immunoprecipitation and western blot analysis. CCL2-induced phosphorylation of MAP kinases and monocyte migration was significantly attenuated by APN in vitro . Conclusion: The opposite association of APN and CCL2 on the percentage of coronary plaque might be caused by their direct interaction and competitive functions on monocyte migration.

Anti-Inflammatory Effect of KW-2449 on Autoimmune Encephalomyelitis: An Experimental Study on Mice.

The KW-2449 is a novel multikinase inhibitor that inhibits FLT3, ABL, ABL-T315I, and Aurora A. FLT3 and Aurora A kinases play an important role in the pathogenesis of multiple sclerosis (MS). KW-2449 could modulate immune cells, but the immunomodulatory effects of KW-2449 on experimental autoimmune encephalomyelitis (EAE) have not been investigated yet. The aim of the present study is to investigate the effects of KW-2449 on EAE mouse model. In this study, C57BL/6 EAE mice were orally treated with (10 mg/kg/day) KW-2449 solution and compared with EAE and control mice. Following the treatment, histological analyses were performed on the brain and cerebellum to evaluate the pathological score. The gene expression levels of tumor necrosis factor-alpha (TNF-α), interleukin 6 (IL-6), and chemokine (C-C motif) ligand 2 (CCL2) were measured using qRT-PCR. The serum levels of TNF-α, IL-6, CCL-2 and MMP-2 were determined by using quantitative enzyme-linked immunosorbent assay (ELISA). The results indicated that the clinical score, the infiltration of inflammatory cells and the demyelination in EAE mice treated with KW-2449 decreased significantly compared to control groups. KW-2449 also decreased TNF-α, IL-6, CCL-2 inflammatory cytokines, and MMP-2 in both brain mRNA expressions and serum levels of EAE mice. The KW-2449, aging as a multi-kinase inhibitor, modulates the inflammatory responses of cytokine cascades either in the brain or in plasma and reduces EAE pathogenesis manifestation.

Serum levels of CC chemokine ligands in cutaneous leishmaniasis patients

The crucial functions of chemokine/receptors in numerous parasitic infections, including leishmaniasis, are well documented. This study aimed to assess the serum levels of CC ligand (CCL) 2, CCL5, and CCL11 in cutaneous leishmaniasis (CL) patients. 64 patients, suffering from CL and 100 healthy people were selected, and their blood serum concentrations of CCL2, CCL5, and CCL11 were measured using enzyme-linked immunosorbent assay. The results demonstrated that while the mean serum levels of CCL5 and CCL11 increased significantly in CL patients, the mean serum levels of CCL2 decreased, compared to the control group. Despite the sufficient production of CCL5 and CCL11 in CL patients, they suffered from CCL2 deficiency, as the defense mechanism against parasitic infection. These findings suggest a mechanism that might partially explain the patients' susceptibility to persistent infection and their inability to clear the parasites.

Cytokines CCL2 and CXCL1 may be potential novel predictors of early bone loss.

Osteoporosis is a common disorder characterized by decreased bone mineral density (BMD) and increased fracture risk. The current techniques detect real-time BMD precisely but do not provide adequate information to predict early bone loss. If bone loss could be diagnosed and predicted early, severe osteoporosis and unexpected fractures could be prevented, allowing for an improved quality of life for individuals. In the present study, an ovariectomized rat model of bone loss was established and the serum levels of 78 potential cytokines were determined using a protein array. The BMD of ovariectomized rats was dynamically measured by micro-CT and the early stage of bone loss was defined at the fourth week after surgery. The expression of several serum protein cytokines was indicated to be altered in the ovariectomized rats during an 8-week time-course of bone loss. Linear regression analysis revealed that the serum levels of C-C motif chemokine ligand 2 (CCL2, also known as monocyte chemoattractant protein 1) and C-X-C motif chemokine ligand 1 (CXCL1) were significantly associated with a reduction in BMD. The significance of these two factors in indicating bone mass reduction was further verified by analyzing serum samples from 24 patients with BMD using ELISA and performing a linear regression analysis. The serum levels of CCL2 and CXCL1 were inversely correlated with the bone mass. Therefore, the cytokines CCL2 and CXCL1 may be potential novel predictors of early bone loss and may be clinically relevant for the early diagnosis and prevention of osteoporosis.

CCL11 levels in drug-naive bipolar patients: The role of sex and smoking status

• Changes in CCL11 levels may not be involved in the early stages of bipolar disorder. • Demographic and lifestyle factors may drive changes in CCL11 levels in young adults. • Tobacco use and male sex may be associated with higher CCL11 levels in young adults. It has been reported that patients with bipolar disorder (BD) present changes in peripheral levels of the inflammatory cytokine CCL11, particularly at latestages. The aim of this study is to evaluate CCL11 levels in a population-based sample of drug-naive young adults. This is a cross-sectional study nested to a large population-based study. 29 drug-naive young adults with BD and 29 controls selected from this cohort were matched for sex, age, and years of education. The diagnosis of psychiatric disorders was performed using the Mini International Neuropsychiatric Interview PLUS. Serum levels of CCL11 were measured using sandwich-ELISA. Independent samples T-test was used to assess differences between groups regarding CCL11 levels. Multivariate linear regression analyses were used to evaluate the effect of independent factors on CCL11 levels. There were no demographic differences between individuals with BD and controls. No significant differences were found regarding CCL11 levels between groups. The final multivariate model showed that the variables that remained independently associated with higher CCL11 levels were male sex (B:41.41 [CI95%: 15.66 - 67.15], p = 0.002) and tobacco use (B:22.71 [CI95%: 1.28 - 44.15], p = 0.038). The present study suggests that there is a possible influence of sex and tobacco use on CCL11 levels, as male sex and smoking status were associated with higher CCL11 levels in our sample. It also suggests that peripheral levels of CCL11 may not be involved in the pathophysiology of BD at early stages of the disorder.

PNPLA3 rs738409 associates with alcoholic liver cirrhosis but not with serum levels of IL6, IL10, IL8 or CCL2 in the Russian population

Introduction and aimPolymorphic variant rs738409 within the PNPLA3 gene associates with alcoholic liver cirrhosis (ALC) in heavy drinkers of various ancestry but has not yet been established in the Russian population characterized by high incidence of ALC. PNPLA3 rs738409 involvement in the inflammatory process has been proposed as one of the mechanisms of liver dysfunction. Relationship between the PNPLA3 polymorphism and the biochemical markers of inflammation in patients with ALC remains unclear. The current study revealed the association between the rs738409 polymorphism, liver cirrhosis and serum cytokines in heavy drinkers in the Russian population. Materials and methodsThe serum levels of IL6, IL10, IL8, and CCL2 along with PNPLA3 rs738409 polymorphism were determined in heavy drinkers (AA, n=71) and heavy drinkers with diagnosed liver cirrhosis (ALC, n=110). All of the recruited individuals were Caucasians and belonged to the Russian population. ResultsHeavy drinkers carrying PNPLA3 rs738409 CG or CG+GG genotypes as compared with CC genotype carriers or G allele as compared with C allele carriers had significant risk of ALC. In ALC levels of interleukins and CCL2 increased as compared with AA. PNPLA3 rs738409 CC carriers had lower cirrhosis stage as compared with CG+GG carriers, however there were no differences of IL6, IL10, IL8 or CCL2 levels between G allele carriers and non-carriers in heavy drinkers. ConclusionThus, in the Russian population heavy drinkers carrying PNPLA3 rs738409 G allele are at higher risk of ALC, however the presence of rs738409 allele does not influence the serum cytokine levels.

Serum CCL22 levels decreased in parallel with disease activity in CCR4-positive mycosis fungoides treated with mogamulizumab.

Mogamulizumab is a humanized anti-C-C chemokine receptor type (CCR)4 antibody that shows cytotoxicity against CCR4+ lymphoma cells via antibody-dependent cell-mediated cytotoxicity in advanced cutaneous T cell lymphoma (CTCL) patients. The production levels of ligands for CCR4, that is, Chemokine (C-C motif) ligand (CCL)17 and CCL22, are important for the assessment of the disease activity in CTCL patients. We evaluated the serum levels of CCL17, CCL19, CCL22, C-X-C motif chemokine ligand (CXCL)10, and CXCL13, which are ligands for CCR4, CCR7, CCR4, C-X-C Motif Chemokine Receptor (CXCR)3, and CXCR5, respectively, at baseline and 4 weeks after the administration of mogamulizumab in five patients with mycosis fungoides. The serum levels of CCL22 were significantly decreased in patients who responded to mogamulizumab, but no differences were identified in the serum levels of CCL17, CCL19, CXCL10, or CXCL13. Immunofluorescence staining revealed that the majority of CCL22-producing cells were cluster of differentiation (CD)163+ tumor-associated macrophages, and they were surrounded by CCR4+ CTCL cells. Our present data suggested that the serum CCL22 level may be a predictive marker of the efficacy of mogamulizumab for the treatment of CCR4+ CTCL.

Immunomodulatory drugs suppress Th1-inducing ability of dendritic cells but enhance Th2-mediated allergic responses

AbstractImmunomodulatory drugs (IMiDs), lenalidomide and pomalidomide, are widely used treatments for multiple myeloma; however, they occasionally lead to episodes of itchy skin and rashes. Here, we analyzed the effects of IMiDs on human myeloid dendritic cells (mDCs) as major regulators of Th1 or Th2 responses and the role they play in allergy. We found that lenalidomide and pomalidomide used at clinical concentrations did not affect the survival or CD86 and OX40-ligand expression of blood mDCs in response to lipopolysaccharide (LPS) and thymic stromal lymphopoietin (TSLP) stimulation. Both lenalidomide and pomalidomide dose-dependently inhibited interleukin-12 (IL-12) and TNF production and STAT4 expression, and enhanced IL-10 production in response to LPS. When stimulated with TSLP, both IMiDs significantly enhanced CCL17 production and STAT6 and IRF4 expression and promoted memory Th2-cell responses. In 46 myeloma patients, serum CCL17 levels at the onset of lenalidomide-associated rash were significantly higher than those without rashes during lenalidomide treatment and those before treatment. Furthermore, serum CCL17 levels in patients who achieved a very good partial response (VGPR) were significantly higher compared with a less than VGPR during lenalidomide treatment. The median time to next treatment was significantly longer in lenalidomide-treated patients with rashes than those without. Collectively, IMiDs suppressed the Th1-inducing capacity of DCs, instead promoting a Th2 response. Thus, the lenalidomide-associated rashes might be a result of an allergic response driven by Th2-axis activation. Our findings suggest clinical efficacy and rashes as a side effect of IMiDs are inextricably linked through immunostimulation.

Autocrine CCL5 Effect Mediates Trastuzumab Resistance by ERK Pathway Activation in HER2-Positive Breast Cancer.

HER2-positive breast cancer is currently managed with chemotherapy in combination with specific anti-HER2 therapies, including trastuzumab. However, a high percentage of patients with HER2-positive tumors do not respond to trastuzumab (primary resistance) or either recur (acquired resistance), mostly due to molecular alterations in the tumor that are either unknown or undetermined in clinical practice. Those alterations may cause the tumor to be refractory to treatment with trastuzumab, promoting tumor proliferation and metastasis. Using continued exposure of a HER2-positive cell line to trastuzumab, we generated a model of acquired resistance characterized by increased expression of several cytokines. Differential gene expression analysis indicated an overexpression of 15 genes, including five different chemokines, and highlighting CCL5/RANTES as the most overexpressed one. Functional studies, either by in vitro gene silencing or by in vitro and in vivo pharmacologic inhibition of the CCL5/CCR5 interaction with maraviroc, confirmed that CCL5 overexpression was implicated in acquired resistance to trastuzumab, which was mediated by ERK activation. In patient samples, increased CCL5 expression significantly correlated with lower rates of complete response after neoadjuvant therapy, confirmed by detection of high serum CCL5 levels by ELISA. Overexpression of CCL5 correlated with ERK phosphorylation in tumor cells and was statistically associated with worse disease-free survival and overall cancer survival in patients with early HER2-positive breast cancer.

Differential role of lipoteichoic acids isolated from Staphylococcus aureus and Lactobacillus plantarum on the aggravation and alleviation of atopic dermatitis

Lipoteichoic acid (LTA), a cell wall component of gram-positive bacteria, up-regulates inflammatory cytokine production through the toll-like receptor 2 (TLR2) signaling pathway, and also contributes to anti-inflammatory responses against immune cells stimulated by lipopolysaccharides. In the current study, we examined the effects of LTAs isolated from Staphylococcus aureus (aLTA) and Lactobacillus plantarum (pLTA) on the aggravation and alleviation of atopic dermatitis (AD). aLTA strongly induced CCL2 production in THP-1 cells. CCL2 was regulated by the TLR2 pathway including the activation of IRAK2, NF-κB and JNK. CCL2 induced Th2 polarization of CD4+T cells through induction of interleukin (IL)-2, −4, and −5 and inhibition of interferon-gamma (IFN-γ). CCL2 levels and immunoglobulin E (IgE) production were increased in aLTA-injected mice. On the other hand, pLTA moderately affected CCL2 production and it inhibited aLTA-mediated CCL2 production. The serum levels of CCL2 and IgE were inhibited by pLTA pre-injection followed by aLTA reinjection, which resulted in the alleviation of irritant contact dermatitis (ICD) symptoms. Our results suggest that S. aureus infection causes an increase in CCL2 production, and may exacerbate atopic dermatitis (AD)-like symptoms through the excessive IgE production. Alternatively, pLTA alleviated AD-like symptoms by inhibiting aLTA-induced CCL2 and IgE production.

Increased expression of in situ IL-31RA and circulating CXCL8 and CCL2 in pemphigus herpetiformis suggests participation of the IL-31 family in the pathogenesis of the disease.

Pemphigus herpetiformis (PH) is a rare clinical subtype of pemphigus with the presence of urticarial plaques, severe pruritus, rare acantholysis and eosinophilic spongiosis. The aim of this study was to investigate the influence of IL-31 and pro-inflammatory cytokines/chemokines in the pathogenesis of PH. Twenty-five patients with PH and three groups: pemphigus foliaceus (PF=14), pemphigus vulgaris (PV=15) and healthy controls (HC=20) were selected for this study. The groups were analysed by immunohistochemistry utilizing IL-31, IL-31RA, IL-4, IL-17 and TNF-α antibodies. Serum levels of IL-4, IL-13, TNF, CXCL8, CCL5 and CCL2 were evaluated by cytometric bead array. Analysis of IL-31 family of PH patients revealed the following findings: (i) Enhanced in situ expression of IL-31 in PH samples, compared to PF and to PV (epidermis); (ii) Cutaneous IL-31RA expression in PH samples was higher than in PF, PV and HC groups (epidermis and dermis); (iii) PF patients that evolved to PH showed significant increased IL-31RA epidermal expression during the PH phase. Profile of pro-inflammatory cytokines (IL-4, IL-17 and TNF-α) in PH patients' skin exhibited: (i) Enhanced IL-4 expression, when compared to patients with PF (epidermis and dermis) and with PV (epidermis); (ii) Augmented IL-17 expression than PF and PV patients (epidermis); (iii) Augmented expression of TNF-α when compared to PF at the epidermal level. Evaluation of circulating cytokines and chemokines showed higher levels of CXCL8 and CCL2 in PH sera compared to HC group. IL-31 and IL-31RA, cytokines related to pruritus, and pro-inflammatory chemokines (CXCL8 and CCL2) seem to exert a role in the pathogenesis of PH. These findings support future studies to clarify the role of IL-31 pathway as a potential therapeutic target for patients with PH.

BAL and serum multiplex lipid profiling in idiopathic pulmonary fibrosis and fibrotic hypersensitivity pneumonitis

BackgroundDifferential diagnosis between IPF and fibrotic HP (fHP) can be challenging: these two ILDs share many common features but call for different therapeutic approaches. In the present study, differential lipid mediator profiles were analysed by a new method in BAL and serum from HP and IPF patients. Materials and methods76 patients were enrolled retrospectively in the study. Median age (IQR) was 67 years (51–74); 63% were males, 30 had fHP and 46 had IPF. Serum and BAL samples were collected at initial diagnosis. For quantification of serum and BAL lipid mediators was used bead-based multiplex LEGENDPlex™ analysis (Biolegend). ResultsSerum Apo A1 levels were significantly higher in IPF than fHP patients (p = 0.314); indeed, serum levels of CCL2 and Apo C3 were lower in HP than in IPF patients (p = 0.013 and p = 0.041, respectively). BAL concentrations of Apo A1, adipsin, Apo C3 and APN were significantly lower in IPF than in fHP patients (p < 0.0001, p < 0.0001, p = 0.007 and p = 0.023, respectively). In the logistic regression, IPF was tested as dependent variable. Serum levels of Apo A1, CCL2 and Apo C3 were tested as independent variables and ROC curve analysis of model performance showed AUC 93% (p < 0.0001); on the other hand, BAL concentrations of Apo A1, adipsin, Apo C3 and APN showed AUC 81% (p < 0.0001). DiscussionLipid biomarkers evaluated in BAL in our study confirm the hypothesis that fHP and IPF have different lung fibrosis phenotypes. The former is a post-inflammatory cell-regulated ILD and the second is more related to tissue remodeling and repair.

Intrathyroid injection of dexamethasone inhibits Th2 cells in Graves' disease.

Intrathyroid injection of dexamethasone (IID) was used for decrease the relapse rate of hyperthyroidism in the treatment of Graves' disease (GD), but the mechanism is still unclear. We aimed to explore the effect of IID on T help (Th)1/Th2 cells and their chemokine in patients with GD. A total of 42 patients with GD who were euthyroidism by methimazole were randomly divided into IID group (n = 20) and control group (n = 22). Thyroid function and associated antibody, Th1/Th2 cells proportion, serum CXCL10 and CCL2 levels, and CXCR3/CCR2 mRNA expression in peripheral blood mononuclear cells before and after 3-month IID treatment were tested by chemiluminescence assay, Flow cytometry, ELISA, and real-time PCR, respectively. Thyroid follicular cells were stimulated by IFN-γ and TNF-α and treated with dexamethasone in vitro. CXCL10 and CCL2 levels in supernatant were determined. After 3-month therapy, the proportion of Th2 cells and serum CCL2 levels, as well as TPOAb, TRAb levels and thyroid volume decreased in IID group (p < 0.05). However, the proportion of Th1 and CXCL10 levels had no change in IID group and control (p > 0.05). The CXCR3/CCR2 ratio had no change in both groups (p > 0.05). IID therapy could inhibit peripheral Th2 cells via decreasing CCL2 level in peripheral blood, and this result partly explain the effects of IID therapy on prevention of relapse of GD. Arch Endocrinol Metab. 2020;64(3):243-50.

P1709THE ROLE OF CXCL9 AND CXCL13 IN PREDICTION OF POSTTRANSPLANT INFECTION IN KIDNEY TRANSPLANT RECIPIENTS

Abstract Background and Aims Chemokines are deeply involved in the process of inflammatory and immune responses. Interferon-γ-inducible chemokines C-X-C motif chemokine 9 and 10 (CXCL9 and CXCL10) are significantly associated with Th1 cells and monocytes and rise rapidly during early episode of renal allograft rejection and various infectious diseases. CXCL13 is one of the most potent B cells and T follicular helper (Tfh) cells chemoattractants when acts through its cognate receptor CXCR5. Recent work of CXCL13 indicated a critical immune regulatory role in both multiple infectious diseases and kidney transplantation. Additionally, C-C motif chemokine 2 (CCL2) is shown to be is critical for chronic kidney diseases. The aim of this study was to identify the predictive role of serum CXCL9, CXCL10, CXCL13 or CCL2 on kidney posttransplant infection. Method 95 kidney transplant recipients (KTRs) were enrolled in this study. 31 recipients experienced at least once infection episodes within the first posttransplant 12 months and 64 KTRs did not experience any infection episode during the follow-up period. Serum CXCL9, CXCL10, CXCL13 and CCL2 at the time points of pre-transplantation and post-transplant 30 days (POD 30) were analyzed with Bio-Plex® suspension array system. Results It was found that serum level of POD 30 CXCL9 and POD 30 CXCL13 was associated with infection within one year after transplantation (P=0.021, P=0.002, respectively, shown in Figure 1). The serum level of CXCL9 and CXCL13 before surgery, the serum level of CCL2 and CXCL10 before and after surgery were not associated with infection within posttransplant one year (P&amp;gt;0.05, shown in Figure 1). The combination of POD 30 CXCL9 plus POD 30 CXCL13 provided the best results with AUC of 0.721 (95%CI, 0.591-0.852), sensitivity of 71.4% and specificity of 68.5% at the optimal cut-off value of 52.72 pg/ml (shown in Figure 2). Conclusion Chemokines CXCL9 and CXCL13 as important chemokines could be used to predict the occurrence of infection within posttransplant one year in kidney transplant recipients.

CXCL9 and CXCL10 display an age-dependent profile in Chagas patients: a cohort study of aging in Bambui, Brazil

BackgroundChagas disease is endemic in Latin America and still represents an important public health problem in the region. Chronic cardiomyopathy is the most significant chronic form due to its association with morbidity and mortality. The last decade has seen increasing evidence that inflammatory cytokines and chemokines are responsible for the generation of inflammatory infiltrate and tissue damage, with chronic chagasic cardiomyopathy patients presenting a pro-inflammatory immune response. Although studies have evaluated the role of chemokines in experimental T. cruzi infection, few have addressed their systemic profile, especially for human infection and in aging populations. The present work aimed to use the data from a large population based study of older adults, conducted in an endemic area for Chagas disease, to examine the association between serum levels of cytokines and chemokines, T. cruzi infection and electrocardiogram (ECG) abnormality.MethodsThe present work evaluated serum levels of CCL2, CXCL9, CXCL10, CCL5, CXCL8, IL-1β, IL-6, TNF, IL-12 and IL-10 by Flow Cytometric Bead Array assay (CBA) and the results expressed in pg/ml. The baseline survey started in January 1st 1997, with 1284 participants of an aged population-based cohort. Participants signed an informed consent at baseline and at each subsequent visit and authorized death certificate and medical records verification.ResultsOur results demonstrated that Chagas disease patients had higher serum levels of CXCL9, CXCL10 and IL-1β and lower serum levels of CCL5 than non-infected subjects. Moreover, our data demonstrated that CXCL9 and CXCL10 increased in an age-dependent profile in Chagas disease patients.ConclusionTogether, this study provided evidences that serum biomarkers increase along the age continuum and may have potential implications for establishing clinical management protocols and therapeutic intervention in Chagas disease patients.

Evaluation of the Human Pulmonary Activation-Regulated Chemokine (CCL18/PARC) and Alkaline Phosphatase (ALP) Levels in Iraqi Patients with Rheumatoid Arthritis

Rheumatoid arthritis (RA) is one of the chronic inflammatory autoimmune diseases which occurs as a result of unknown reasons. This study was conducted at Baghdad Teaching Hospital/City of Medicine, where blood samples were taken from 60 Iraqi patients with RA (49 females and 11 males) and these patients were matched by age and sex with 20 healthy controls (16 females and 4 males). Patients with RA were diagnosed by a consultant rheumatologist according to ACR / EULAR criteria in 2010. In this study the patients were divided into four groups as follows; the first group consisted of 12 patients treated with methotrexate (MTX), the second group consisted of 10 patients treated with etanercept, the third group consisted of 18 patients treated with a combination of MTX, etanercept and prednisolone, the fourth group consisted of 20 patients treated with MTX and etanercept. Enzyme linked immunosorbent assay (ELISA) was used to detect CCL18/PARC antibodies, while a spectrophotometer (Humalyzer2000) was used for the measurement of alkaline phosphatase (ALP). Serum levels of CCL18 / PARC showed a significant increase in RA patients compared with healthy controls (p ≤ 0.001). The levels of CCL18/PARC showed a significant correlation with disease activity (CDAI), except in RA patients treated with etanercept. There was also no significant correlation between CCL18/PARC and erythrocyte sedimentation rate (ESR). The results showed a significant increase in serum levels of alkaline phosphatase (ALP) was recorded in RA patients (with treatment) as compared to healthy controls (p ≤ 0.001).

Major Depression in Children with Transfusion-Dependent Thalassemia Is Strongly Associated with the Combined Effects of Blood Transfusion Rate, Iron Overload, and Increased Pro-inflammatory Cytokines.

Beta-thalassemia major patients are treated with repeated blood transfusions, which may cause iron overload, which in turn may induce immune aberrations, and show an increased risk of depression. The aim of the present study is to examine whether repeated blood transfusions, iron overload, and immune-inflammatory responses are associated with depression in children (6-12years) with transfusion-dependent thalassemia (TDT). The Children's Depression Inventory (CDI), iron status (serum iron, ferritin, transferrin, TS%), and serum levels of CCL11, IL-1β, IL-10, and TNF-α were measured in TDT with (n = 54) and without (n = 57) a major depression-like episode (MDLE) and in healthy children (n = 55). The results show that MDLE due to TDT is associated with a greater number of blood transfusions and increased iron overload and IL-1β levels. Partial least squares path analysis shows that 68.8% of the variance in the CDI score is explained by the number of blood transfusions, iron overload, and increased levels of IL-1β and TNF-α. The latter two cytokines partly mediate the effects of iron overload on the CDI score, while the effects of blood transfusions on the CDI score are partly mediated by iron overload and the path from iron overload to immune activation. Iron overload is also associated with increased IL-10 and lower CCL11 levels, but these alterations are not significantly associated with depression. In conclusion, blood transfusions may be causally related to MDLE in TDT children and their effects are in part mediated by increased iron overload and the consequent immune-inflammatory response. The results suggest that effects of iron overload and its consequences including inflammation and oxidative stress toxicity may cause MDLE. Current treatment modalities with folic acid and vitamin C are insufficient to attenuate iron overload and immune-inflammatory responses and to prevent MDLE in children with TDT.

In-vivo inducible tissue-specific deletion of chemokine receptor-2 in mouse cartilage decreases joint damage during injury-induced osteoarthritis

Purpose: Increased expression of chemokines in cartilage, synovial membrane and subchondral bone are believed to be linked to osteoarthritis (OA). CCR2 has been recognized as an important potential target in OA: CCR2, its ligands CCL12 (a.k.a. MCP-5) and CCL2 (a.k.a. MCP-1, the major human circulating ligand), are significantly increased in both rodent models of OA and in humans with OA. In addition, they have been shown to mediate OA pain. In previous clinical studies, we determined that CCL2 serum levels at baseline were significantly associated with radiographic knee OA progression and joint space narrowing at 5-year follow up. Using the destabilization of medial meniscus (DMM) murine model of injury-induced OA, we demonstrated that targeting the CCR2 signaling at early OA stages had beneficial effects on OA progression and that there was a disconnect between structure and pain that has been noted in human OA. We therefore hypothesize that alterations in joint mechanics upregulate the CCR2 pathway in cartilage and bone in a time-dependent manner altering the structural organization of both tissues, ultimately leading to OA and pain. In the present study, by using a temporal and tissue specific inactivation of CCR2 in cartilage, we determine the role of CCR2 signaling in-vivo at sequential times during injury-induced OA in order to establish the contribution of CCR2 to the development of whole joint degeneration. Methods: We obtained an inducible tissue-specific CCR2 deletion in chondrocytes by combining chondrocyte-specific expression of CreER with Tamoxifen (Tam) injections in CCR2flx/flxeGFP mice crossed with Aggrecan-CreERT2 (CCR2AggERKO). The Cre-mediated recombination deletes CCR2 exon3, generating a functional knockout, and activates the expression of eGFP under the CCR2 locus. Therefore, recombination can be validated by GFP expression. CCR2AggERKO and CCR2flx/flx mice were injected with Tamoxifen (40μg/g) for 5 days and subjected to DMM 1 week after the last injection. Mice were euthanized at 2, 4, 8 and 12 weeks after DMM and knees were prepared for histological evaluation (Safranin-O/Fast Green, H&E) and immunohistochemical (IHC) analyses with anti-GFP, anti-Collagen10 and anti-osterix antibodies). Results: We successfully inactivated the CCR2 receptor in the cartilage tissue, as shown by the intense GFP expression in the cartilage tissues of CCR2AggERKO mice compared to controls. The expression of CCR2 in other tissues is not altered by the recombination. Histological evaluation shows that deletion of CCR2 in cartilage tissue decreases articular cartilage structure score and Safranin-O staining score at 4, 8 and 12 weeks post-surgery compared to controls. In addition, at both 8 and 12 weeks post surgery, protein levels of Collagen 10 are significantly higher in DMM-CCR2AggERKO mice while almost absent in DMM-controls, were the whole cartilage is replaced by bone. Conclusions: Our results show that we were able to selectively inhibit the CCR2 expression in chondrocytes during OA progression, while its expression was unchanged in other joint tissues. We demonstrated that the CCR2 deletion in cartilage slows cartilage damage at the mild, moderate and severe OA stage compared to controls. Other studies are currently in progress in our lab to determine whether CCR2 deletion in cartilage indirectly affect the bone tissue as well. Behavioral pain analyses are also in progress to determine whether CCR2 cartilage deletion alleviate pain responses.

Nuclear Receptor Small Heterodimer Partner Inhibits Proinflammatory Macrophage Polarization in Liver Disease

ObjectiveMacrophages are the first responding cells to hepatocyte injuries and play a central role in innate immune response during the pathogenesis of liver diseases. Macrophage polarization to pro‐inflammatory M1 or anti‐inflammatory M2 sub‐populations is a critical event of inflammatory response which balances tissue injury and repair. Various types of signaling pathways have been identified that direct macrophage polarization; however, the regulatory mechanisms that coordinate the pathways are still not well understood. In this study we investigated the role of small heterodimer partner (Shp), an orphan nuclear receptor, in regulating the signaling pathways involved in macrophage polarization during the development of liver diseases.MethodsPrimary macrophages isolated from mouse liver were differentiated into pro‐inflammatory M1 or anti‐inflammatory M2 macrophages and Shp expression was determined during the differentiation. Mouse macrophage cell line RAW 264.7 overexpressed with Shp was treated with lipopolysaccharide (LPS) to evaluate inflammatory response. For the in vivo studies, we generated macrophage‐specific Shp deficient mice (Shp‐MKO represents Shp flox/flox; LysMCre+/−) and littermate controls (Shp‐WT represents Shp flox/flox; LysMCre−/−). Two different animal models were used to assess inflammatory responses in the liver, including high fat, cholesterol, and fructose (HFCF) diet‐induced nonalcoholic steatohepatitis (NASH) model and LPS‐induced acute inflammatory model.ResultsShp expression was down‐regulated during pro‐inflammatory M1 macrophage differentiation and up‐regulated in anti‐inflammatory M2 macrophage differentiation. Overexpression of Shp in Raw 264.7 cells resulted in inhibition of LPS‐induced activation of NF‐κB pathway compared to control cells. In addition, Shp overexpression downregulated MAP kinase pathway activation by inhibiting the phosphorylation of MKK4, JNK and c‐jun. Consistently, the in vivo study showed that both NF‐κB and MAP kinase signaling pathways were highly activated in the Shp‐MKO with the increased serum levels of TNF‐α and CCL2 after LPS treatment. Strikingly, macrophage polarization toward pro‐inflammatory M1 stage was enhanced in the Shp‐MKO liver in comparison to the Shp‐WT, which is evidenced by increased iNOS, IL‐6 and TNF‐α production upon LPS injection. On the molecular level we found that SHP interacted with proteins involved in NF‐κB and MAP kinase pathways and thereby coordinated macrophage polarization. Finally, without affecting hepatocyte steatosis Shp‐MKO mice were more susceptible to HFCF‐induced NASH progression with monocyte infiltration and liver fibrosis.ConclusionOur study uncovered a unique role of SHP in regulating macrophage polarization that couples both NF‐κB and MAP kinase pathways, which impacts the pathogenesis of acute and chronic inflammatory liver diseases.Support or Funding InformationThis project was supported by grants NCI K22CA184146, R01DK119131 and NIH P20 GM103549.

Elevated serum chemokine CCL22 levels in first-episode psychosis: associations with symptoms, peripheral immune state and in vivo brain glial cell function

Several lines of research support immune system dysregulation in psychotic disorders. However, it remains unclear whether the immunological marker alterations are stable and how they associate with brain glial cell function. This longitudinal study aimed at investigating whether peripheral immune functions are altered in the early phases of psychotic disorders, whether the changes are associated with core symptoms, remission, brain glial cell function, and whether they persist in a one-year follow-up. Two independent cohorts comprising in total of 129 first-episode psychosis (FEP) patients and 130 controls were assessed at baseline and at the one-year follow-up. Serum cyto-/chemokines were measured using a 38-plex Luminex assay. The FEP patients showed a marked increase in chemokine CCL22 levels both at baseline (p < 0.0001; Cohen’s d = 0.70) and at the 12-month follow-up (p = 0.0007) compared to controls. The group difference remained significant (p = 0.0019) after accounting for relevant covariates including BMI, smoking, and antipsychotic medication. Elevated serum CCL22 levels were significantly associated with hallucinations (ρ = 0.20) and disorganization (ρ = 0.23), and with worse verbal performance (ρ = −0.23). Brain glial cell activity was indexed with positron emission tomography and the translocator protein radiotracer [11C]PBR28 in subgroups of 15 healthy controls and 14 FEP patients with serum CCL22/CCL17 measurements. The distribution volume (VT) of [11C]PBR28 was lower in patients compared to controls (p = 0.026; Cohen’s d = 0.94) without regionally specific effects, and was inversely associated with serum CCL22 and CCL17 levels (p = 0.036). Our results do not support the over-active microglia hypothesis of psychosis, but indicate altered CCR4 immune signaling in early psychosis with behavioral correlates possibly mediated through cross-talk between chemokine networks and dysfunctional or a decreased number of glial cells.