Peripheral Monocytes In Patients Research Articles

Natalizumab Promotes Activation of Peripheral Monocytes in Patients With Multiple Sclerosis.

Natalizumab (NTZ), a monoclonal antibody against very late antigen-4 (VLA-4), is one of the most efficient therapies to prevent acute relapses in multiple sclerosis (MS). VLA-4 is the key adhesion molecule for peripheral immune cells, especially lymphocytes to enter the CNS. While its blockade thus virtually abrogates CNS infiltration of these cells, long-term exposure to natalizumab may also affect immune cell function. In this study, we report that in patients with MS, NTZ treatment is associated with an enhanced activation status of peripheral monocytes. Expression of 2 independent activation markers, CD69 and CD150, was significantly higher on blood monocytes from NTZ-treated patients when compared with those from matched untreated patients with MS, while other properties such as cytokine production remained unchanged. These findings consolidate the concept that peripheral immune cells remain fully competent on NTZ treatment, an excellent asset rare among MS treatments. However, they also suggest that NTZ may exert nondesirable effects on the progressive aspect of MS, where myeloid cells and their chronic activation are attributed a prominent pathophysiologic role.

Inactivation of TMEM106A promotes lipopolysaccharide-induced inflammation via the MAPK and NF-κB signaling pathways in macrophages.

Pattern recognition receptors, such as Toll-like receptors (TLRs), play an important role in the host defense against invading microbial pathogens. Their activation must be precisely regulated, as inappropriate activation or overactivation of TLR signaling pathways may result in inflammatory disorders, such as septic shock or autoimmune diseases. TMEM106A is a type II transmembrane protein constitutively expressed in macrophages. Our current study demonstrated that TMEM106A levels were increased in macrophages upon lipopolysaccharide (LPS) stimulation, as well as in the peripheral monocytes of patients with sepsis. Tmem106a knockout mice were more sensitive to lipopolysaccharide (LPS)-induced septic shock than wild-type mice. Further experiments indicated that Tmem106a ablation enhanced the expression of CD80, CD86 and major histocompatibility complex (MHC)-II in mouse macrophages upon LPS stimulation, accompanied with up-regulation of tumor necrosis factor (TNF)-α, interleukin (IL)-6, interferon (IFN)-β and inducible nitric oxide synthase (iNOS), indicating the activation of macrophages and polarization towards the M1 inflammatory phenotype. Moreover, elevated mitogen-activated protein kinase (MAPK) and nuclear factor kappa B (NF-κB) signaling were found to be involved in the LPS-induced inflammatory response in Tmem106a-/- macrophages. However, this effect was largely abrogated by macrophage deletion in Tmem106a-/- mice. Therefore, deficiency of Tmem106a in macrophages may enhance the M1 polarization in mice, resulting in inflammation. This suggests that TMEM106A plays an important regulatory role in maintaining macrophage homeostasis.

Trimetazidine Improves the Outcome of EECP Therapy in Patients with Refractory Angina Pectoris

Introduction:Cardiovascular disease (CAD) associated with death and disability remains a serious medical problem. In some patients the initial clinical coronary artery disease presentation is stable angina pectoris.Aim:The aim of the study was to evaluate the effect of EECP therapy with or without trimetazidine (TMZ) in patients with refractory angina via modulating peripheral monocyte expression of Toll like receptor2 (TLR2) and its downstream signaling.Methods:This is a double-blind randomized prospective study in which 88 stable refractory angina patients allocated into two groups, Enhanced External Counter Pulsation (EECP) group: included 44 patients with stable refractory angina, and were treated with EECP-Therapy. TMZ-EECP group: included 44 patients with stable refractory angina, we gave TMZ 35 mg twice daily in addition to EECP-Therapy.Results:TLR2 expression in peripheral monocyte investigated by flow cytometry and 8-iso-prostaglandin F2β (8-iso-PGF2 β), interleukin1β (IL-1β), heat shock protein 60 (HSP60) and monocytes chemoattractant protein-1(MCP-1) were also measured before the EECP-therapy and before giving TMZ to patients, and after 35 hours of EECP treatment (7 consecutive weeks). Inhibition in TLR2 expression in peripheral monocyte was observed among the EECP group (P<0.05). Inflammatory cytokine MCP-1 was remarkably decreased in both study groups but (heat shock protein 60 (HSP60), MCP-1 and interleukin-1β (IL-1β)) significantly decreased levels were observed among the TMZ-EECP group (P<0.05). Also, the oxidative stress biomarker 8-iso-prostaglandin F2β (8-iso-PGF2β) was decreased in both study groups but significantly decreased levels were observed among the TMZ-EECP group (P<0.05). TMZ and EECP therapy in patients with stable refractory angina remarkably decreased the inflammatory markers HSP60, MCP-1 and IL-1β in serum levels also the decreased levels were found in serum levels of oxidative stress marker 8-iso-PGF2β serum level.Conclusion:EECP-therapy decreased the expression of TLR2 on peripheral monocytes in patients with chronic stable refractory angina which yield improvement in the quality of patients’ life by decreasing the frequency of angina episodes, decreasing the Short-acting nitrate use and change the exercise tolerance and distance.

P2861Elevated cc chemokine receptor 2 expression and higher migratory activity of monocytes in atrial fibrillation patients with progressive structural remodeling

Abstract Background Inflammation in atrial tissue underlies structural remodeling of left atrium, which is a hallmark of atrial fibrillation (AF). Activated monocytes mediate inflammation; however, the role of monocytes in AF pathogenesis has not been extensively examined. In this study, we thus investigated the association between structural remodeling of left atrium, represented by left atrial dimension (LAD), and characteristics of peripheral monocytes in patients with AF. Methods Blood samples were collected from patients undergone catheter ablation between July 2017 and October 2018, including AF patients (n=152) and paroxysmal supraventricular tachycardia (PSVT) patients, which serves as a control non-AF group (n=22). AF patients were further divided into two groups by the median of LAD (normal LAD group: LAD &lt;40 mm, n=77, large LAD group: LAD ≥40 mm, n=75). Peripheral blood mononuclear cells (PBMC) were isolated to analyze monocyte subsets by flow cytometry. In a subset of patients, we further isolated monocytes from PBMC by using magnetic bead-based negative selection method then gene products associated with inflammation or monocyte functions were evaluated. We also examined migratory activity of monocytes toward monocyte chemotactic protein-1, a ligand for CC chemokine receptor 2 (CCR2), using a modified Boyden chamber method. Finally, we performed immunofluorescence staining of monocytes and macrophages in left atrial appendages resected from patients underwent coronary bypass graft surgery (CABG) complicated by AF. Results There were no differences in age, body mass index and high-sensitivity C-reactive protein levels among three groups, including non-AF, normal LAD and large LAD groups, except that more female subjects were included in non-AF group. We found that proportions of classical CD14++CD16- and nonclassical CD14+CD16++ monocytes were higher (non-AF: 71.2±7.3% vs. AF: 75.5±8.3%, p&lt;0.05) and lower (non-AF: 16.4±5.9% vs. AF: 13.2±5.5%, p&lt;0.05), respectively, in all AF patients compared with those in non-AF group, while no significant difference was observed between normal and large LAD groups. In monocytes from large LAD group, mRNA levels of CCR2, a receptor to mediate monocyte chemotaxis, were significantly higher compared to those in normal LAD group (Figure A, p&lt;0.05). Furthermore, monocytes isolated from large LAD group exhibited higher migratory capacity compared to normal LAD group (Figure B, p&lt;0.01). Finally, higher monocyte/macrophage infiltrations to left atrial appendages were implicated in patients with large LAD, shown by immunofluorescence staining. Conclusions Monocytes in AF patients with enlarged left atrium expressed higher CCR2 mRNA and were more active in chemotaxis to MCP-1, suggesting the proactive roles of activated monocytes in the pathogenesis of arterial remolding in AF.

Elevated expression levels of the voltage-gated sodium channel 1.7 and BAFF receptor in peripheral monocytes contribute to activation of monocytes of patients with primary Sjögren’s syndrome

Abstract Background &amp; Purpose We reported that elevated expression of BAFF-receptor (BR3) in peripheral monocytes of patients with primary Sjögren’s syndrome (pSS) resulted in robust increased in IL-6 production by the cells. It has been reported that the pathways via voltage-gated sodium channels, such as Nav1.7, in monocytic lineage are involved in production of inflammatory cytokines, such as IL-6, in inflammatory pain diseases. In this study, we investigated the expression levels of Nav1.7 and BR3 in peripheral monocytes in patients with pSS in an attempt to discriminate the disease from other autoimmune diseases, such as active RA and active SLE. Methods The expression levels of Nav1.7 and BR3 in peripheral monocytes were analyzed by FACS using whole blood samples from patients with pSS (n = 28), active RA (n = 15), active SLE (n = 37) and healthy controls (HC; n = 15). Peripheral monocytes were stimulated with BAFF in the presence or absence of an inhibitor against Nav1.7. The amount of IL-6 in the culture supernatants was measured by ELISA. Results FACS analysis revealed that the expression levels of Nav1.7 and BR3 in pSS monocytes was significantly higher than those of active RA (p = 0.005, 0.003), active SLE (p &amp;lt;0.001, 0.004) and HC (p &amp;lt; 0.001, 0.002). Interestingly, the expression level of Nav 1.7 in pSS monocytes was significantly and positively correlated with that of BR3 in the cells (p = 0.01). Moreover, a specific inhibitor against Nav 1.7 suppressed IL-6 production by sBAFF-stimulated peripheral monocytes in a dose dependent manner. Conclusion Our results strongly suggest that the crosstalk between BAFF signaling and sodium channel is involved in activation of pSS monocytes and can be a therapeutic target for pSS.

Prognostic significance and population dynamics of peripheral monocytes in patients with oropharyngeal squamous cell carcinoma.

Several inflammatory biomarkers are considered potential prognostic factors in various cancers. This study aimed to investigate the prognostic significance and population dynamics of pretreatment inflammatory biomarker levels in patients with oropharyngeal squamous cell carcinoma (OPSCC). The influence of neutrophil counts, lymphocyte counts, monocyte counts, platelet counts, lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio, and platelet-to-lymphocyte ratio on progression-free survival (PFS), and overall survival (OS) was analyzed. We also analyzed the peripheral blood mononuclear cells collected from patients and healthy donors (HDs). Elevated monocyte count was an independent prognostic factor for PFS. Low LMR was an independent prognostic factor for OS. The proportion of intermediate monocytes was lower, and that of classical monocytes was higher in patients than in HDs. Furthermore, PD-L1 expression on monocytes was higher in patients than in HDs. We showed the prognostic significance and population dynamics of peripheral monocytes in patients with OPSCC.

Abstract 4627: Prognostic significance and population shift of peripheral monocytes in patients with oropharyngeal squamous cell carcinoma

Abstract Evidence has accumulated that chronic inflammation plays a critical role in carcinogenesis, tumor growth, and metastasis. To date, several inflammatory biomarkers are considered as potential prognostic factors in various cancers. The aim of the present study was to investigate the prognostic significance of the pretreatment levels of inflammatory biomarkers in patients with oropharyngeal squamous cell carcinoma (OPSCC). A total of 67 patients newly diagnosed as OPSCC was finally included. The neutrophil, lymphocyte, monocyte and platelet counts recorded before treatment was initiated, then lymphocyte-to-monocyte ratio (LMR), neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) were calculated. These inflammatory biomarkers and clinical parameters including age, differentiation, T-stage, N-stage, TNM-stage, smoking, and drinking were compared with progression-free survival (PFS) and overall survival (OS) using cox regression. Elevated monocyte count and advanced T-stage were significantly associated with poor PFS; in addition, these were independent prognostic factors for PFS. Elevated monocyte count and low LMR were significantly associated with poor OS; moreover, low LMR was an independent prognostic factor for OS. Of note, monocyte count and LMR were associated with prognosis more intensively in patients with p16-negative OPSCC. Following these results, we evaluated the peripheral blood mononuclear cells of 14 patients with OPSCC collected before treatments. CD14+ CD86+ HLR-DR+ cells were gated as monocytes, then divided into three subsets according to the expression of CD14 and CD16. The proportion of intermediate monocytes was lower and that of classical monocytes was higher in OPSCC patients than in healthy donors, especially in patients with elevated monocyte count. Furthermore, the expression level of PD-L1 was higher in patients than in healthy donors on all subsets. In conclusion, our data suggests that elevated monocyte count and low LMR seem to be useful biomarkers for predicting prognosis of OPSCC; furthermore, the population shift of monocytes may relate to poor prognosis of OPSCC. Citation Format: Hideyuki Takahashi, Koichi Sakakura, Kazuaki Chikamatsu. Prognostic significance and population shift of peripheral monocytes in patients with oropharyngeal squamous cell carcinoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 4627. doi:10.1158/1538-7445.AM2017-4627

Characterization of Gene Expression Phenotype in Amyotrophic Lateral Sclerosis Monocytes

Amyotrophic lateral sclerosis (ALS) is a common adult-onset neurodegenerative disease characterized by selective loss of upper and lower motor neurons. Patients with ALS have persistent peripheral and central inflammatory responses including abnormally functioning T cells and activated microglia. However, much less is known about the inflammatory gene profile of circulating innate immune monocytes in these patients. To characterize the transcriptomics of peripheral monocytes in patients with ALS. Monocytes were isolated from peripheral blood of 43 patients with ALS and 22 healthy control individuals. Total RNA was extracted from the monocytes and subjected to deep RNA sequencing, and these results were validated by quantitative reverse transcription polymerase chain reaction. The differential expressed gene signatures of these monocytes were identified using unbiased RNA sequencing strategy for gene expression profiling. The demographics between the patients with ALS (mean [SD] age, 58.8 [1.57] years; 55.8% were men and 44.2% were women; 90.7% were white, 4.65% were Hispanic, 2.33% were black, and 2.33% were Asian) and control individuals were similar (mean [SD] age, 57.6 [2.15] years; 50.0% were men and 50.0% were women; 90.9% were white, none were Hispanic, none were black, and 9.09% were Asian). RNA sequencing data from negative selected monocytes revealed 233 differential expressed genes in ALS monocytes compared with healthy control monocytes. Notably, ALS monocytes demonstrated a unique inflammation-related gene expression profile, the most prominent of which, including IL1B, IL8, FOSB, CXCL1, and CXCL2, were confirmed by quantitative reverse transcription polymerase chain reaction (IL8, mean [SE], 1.00 [0.18]; P = .002; FOSB, 1.00 [0.21]; P = .009; CXCL1, 1.00 [0.14]; P = .002; and CXCL2, 1.00 [0.11]; P = .01). Amyotrophic lateral sclerosis monocytes from rapidly progressing patients had more proinflammatory DEGs than monocytes from slowly progressing patients. Our data indicate that ALS monocytes are skewed toward a proinflammatory state in the peripheral circulation and may play a role in ALS disease progression, especially in rapidly progressing patients. This increased inflammatory response of peripheral immune cells may provide a potential target for disease-modifying therapy in patients with ALS.

Abstract 124: Association of AdipoR2 mRNA Expression on Peripheral Monocytes With Carotid Plaque Instability

Adiponectin, an adipose tissue secreted protein, possesses anti-atherosclerotic properties. Its effects are partly mediated through its action on monocytes, via receptors, AdipoR1 and AdipoR2. Circulating monocytes play a crucial role both in plaque initiation and in the progression of atherosclerosis. The aim of this ongoing study is to investigate for the first time whether there is differential expression of AdipoR1/AdipoR2 on human peripheral monocytes in patients with unstable vs. stable carotid atherosclerotic plaques. Patients (n=20) with carotid stenosis of ≥50% scheduled for a carotid endarterectomy (CEA) were recruited from the Royal Victoria Hospital, Montreal, Canada. Blood samples were collected the morning of the CEA. A radioimmunoassay was used for plasma total adiponectin measurement. Circulating monocytes were isolated from mononuclear cells using a Magnetic Cell-Sorting technique with CD14+ Human Microbeads. Expression of AdipoR1/AdipoR2 mRNA on peripheral monocytes was determined using quantitative RT-PCR. Carotid plaque specimens were processed for histological analyses. Plaque instability was categorized according to the gold standard American Heart Association classification. All specimens were graded as either a Type V (n=11) or a Type VI plaque (n=9). Type VI represents a more unstable lesion than Type V. AdipoR2 gene expression on monocytes was inversely associated with greater plaque instability; compared to patients with a Type V plaque, those with a Type VI plaque had 1.88-fold decreased AdipoR2 mRNA levels (p=0.046). AdipoR2 gene expression was found to be a trending predictor of a Type VI plaque (OR=2.93; 95% CI=0.87-9.81; p=0.082). AdipoR1 gene expression on monocytes did not differ between the two groups, while plasma adiponectin levels were lower in Type VI plaques (Type VI: 2.73 [2.19-3.96] μg/ml vs. Type V: 6.97 [4.69-10.07] μg/ml; p= 0.067). Plasma adiponectin levels did not correlate with the expression levels of AdipoR1/AdipoR2 on monocytes. These results indicate that carotid plaque instability is associated with decreased AdipoR2 expression on peripheral monocytes. Our ongoing study may determine the underlying mechanisms linking adiponectin and its receptors with plaque instability.

Differential gene expression of bradykinin receptors 1 and 2 in peripheral monocytes from patients with essential hypertension

Bradykinin participates in various hypertensive processes, exerted via its type 1 and type 2 receptors (BKR1 and BKR2). The aim of the study was to investigate BKR1 and BK2R gene expression in peripheral monocytes in patients with essential hypertension compared with healthy individuals. Seventeen hypertensive patients (9 males, age 56 ± 7 years) and 12 healthy individuals (7 males, age 55 ± 6) participated. Mononuclear cells isolated using anti-CD14+ antibodies and mRNAs of BKR1 and BKR2 were estimated by real-time quantitative reverse transcription-PCR. Both BKR1 and BKR2 showed significantly upregulated gene expression in the group of hypertensive patients. Specifically, BKR1 gene expression was 142.1 ± 42.2 in hypertensives versus 20.2 ± 8 in controls (P = 0.024) and BKR2 was 1222.2 ± 361.6 in hypertensives versus 259.5 ± 99.1 in controls (P = 0.038). Antihypertensive treatment resulted in a decrease in BKR1 (from 142.1 ± 42.2 to 55.2 ± 17.1, P = 0.065) and in BKR2 (from 1222.2 ± 361.6 to 256.8 ± 81.8, P = 0.014) gene expression. BKR1 and BKR2 gene expression on peripheral monocytes is upregulated in essential hypertension. This may lead to functional changes in monocytes and contribute to the development of target organ damage in hypertensive patients.

AB0045 Baff signaling pathway may be involved in the production of matrix metalloproteinase-9 by peripheral monocytes of patients with primary sjögren’s syndrome.

BackgroundB cell activating factor of the TNF family (BAFF) is a type II membrane-bound protein and the extracellular C-terminal fragment is released from the cells as soluble BAFF (sBAFF). BAFF...

FRI0042 Baff signaling is abnormally regulated through JAK pathways in peripheral monocytes in patients with primary sjÖgren’s syndrome

BackgroundB cell activating factor belonging to the TNF superfamily (BAFF) regulates proliferation, differentiation and survival of B cells and plays a pivotal role in the pathogenesis of autoimmune diseases such...

NF-kB is involved in abnormal BAFF signaling in peripheral monocytes of patients with primary Sjögren’s syndrome. (P6274)

Abstract Background and Purpose: We revealed that BAFF induced a robust increase in the production of IL-6 by monocytes from patients with primary Sjögren’s syndrome(pSS), and that the expression levels of a BAFF receptor (BR3) , NF-IL-6 and NF-kB2 were enhanced in pSS monocytes compared to the control. The purpose of this study is to elucidate the possible abnormalities of a BAFF signaling pathway in pSS monocytes. Materials and Methods: The expression level of BR3 on monocytes in whole blood from pSS patients or normal control was analyzed by FACS. Peripheral monocytes were stimulated with soluble BAFF (sBAFF), and the production of IL-6 by the cells was measured by ELISA. The expression levels of IKK-alpha and IKK-beta were analyzed by western blotting. Results and Discussion: FACS analysis showed that the expression level of BR3 was significantly elevated in pSS monocytes compared to the control. Interestingly, the expression level of BR3 on pSS monocytes was positively correlated with the amount of IL-6 produced by sBAFF-treated monocytes. Moreover, NF-kB activation inhibitor suppressed the IL-6 production by sBAFF-treated pSS monocytes in a dose-dependent manner. In addition, phosphorylation levels of IKK-alpha and IKK-beta in pSS monocytes were enhanced upon stimulation with sBAFF. These data suggest that sBAFF acts through BR3 to induce the expression of IL-6 in monocytes, and that IKK-alpha and IKK-beta are involved in the signal transduction pathway triggered by sBAFF.

Surface expression and genotypes of Toll-like receptors 2 and 4 in patients with juvenile idiopathic arthritis and systemic lupus erythematosus

BackgroundChronic arthritis is a common feature of juvenile idiopathic arthritis (JIA) and systemic lupus erythematosus (SLE). It was subsequently discovered that Toll-like receptors (TLRs) are able to upregulate cytokine production in response to endogenous ligands released after tissue damage, suggesting that TLRs can maintain an inflammatory response even in absence of pathogen. Thus, TLRs may contribute to increased inflammation in JIA and SLE patients. The aim of this study was to investigate the role of TLRs in JIA and SLE. We examined the in vivo expression and polymorphisms of TLR2 and TLR4 in peripheral monocytes of patients with JIA and SLE during active and inactive disease phases.MethodsThis single center cohort study consisted of JIA and SLE affected children and control subjects. TLR2 and TLR4 protein expression on CD14+ monocytes was examined by flow cytometry. TLR2 and TLR4 genotypes were determined using the polymerase chain reaction-restriction fragment length polymorphism method (RFLP-PCR).ResultsA significant reduction in the level of TLR4 expression (p ≤ 0.001) was observed on monocytes of patients with JIA and SLE compared with that of healthy control subjects. There was no correlation between the TLR2 or TLR4 genotypes and the observed differential TLR protein expression on monocytes.ConclusionsTo conclude, our observations suggest involvement of investigated TLRs in the pathogenesis of JIA and SLE. It still remains to be elucidated whether reduced TLR4 expression is cause of chronic arthritis or a result of some feedback loop.

Up-Regulation of Toll-Like Receptors 2 and 4 on Peripheral Monocytes After Major Abdominal Surgical Operation

This study is to investigate the time-course expression of TLR2 and TLR4 on peripheral monocytes in patients receiving major abdominal surgical operation. Blood samples were obtained from peripheral vein before and after an operation in 30 patients with gastrointestinal or pancreatic surgery. The mRNA expression of TLR2, TLR4, TNF-α and IL-6 on peripheral blood mononuclear cells (PBMC) was analyzed by real-time PCR. The expressions of TLR2, TLR4, HLA-DR, CD80, and CD86 on monocytes were analyzed by flow cytometry. The expressions of TLR2 and TLR4 on monocytes responding to each agonist (zymosan and lipopolysaccharide) were also measured by flow cytometry. TLR2 and TLR4 mRNA showed significant up-regulation after the completion of the operation when compared with those before the operation. TLR2 expression reached its peak level on day 1 and TLR4 on days 1 and 3. There was no significant difference between pre- and post-operation in the expressions of HLA-DR, CD80 and CD86. Stimulation with zymosan, increased the expression of TLR2 significantly after the operation and reached its highest value on day 3. Similarly, after stimulation with lipopolysaccharide, the expression of TLR4 was also increased and the highest level was observed on day 1. The expression of TNF-α and IL-6 mRNA decreased after completion of the operation and gradually returned to basal level. The expressions of TLR2 and TLR4 on monocytes were up-regulated during the early period after a major abdominal surgical operation in patients, which might be related to the activation of innate immunity.

P081 JAK3 is involved in BAFF-mediated abnormal production of IL-6 by peripheral monocytes of patients with primary Sjögren’s syndrome

Introduction B cell activating factor belonging to the TNF superfamily (BAFF) regulates proliferation, differentiation and survival of B cells. Several lines of evidence have been reported that elevated production of BAFF seemed to be involved in the development of autoimmune diseases such as primary Sjogren’s syndrome (pSS). In our previous study, we revealed that pSS monocytes produced significantly higher amount of IL-6 than normal monocytes upon stimulation with soluble BAFF (sBAFF). These data suggest that the production of IL-6 by monocytes in induced partly through a signal transduction pathway triggered by sBAFF. Since IL-6 may be involved in the production of autoantibodies by B cells, elucidation of the regulatory mechanism of the aberrant production of IL-6 by pSS monocytes is important not only to understand the mechanism of pathogenesis of pSS, but also to explore therapeutic possibilities to treat the disease. In the present study, we investigated possible abnormalities of BAFF signaling pathways in pSS monocytes. Methods The expression levels of BAFF receptor (BAFF-R) on peripheral monocytes of pSS patients (n = 10) and age-matched normal individuals (n = 8) were analyzed by FACS and quantitative PCR. The monocytes were stimulated in vitro with sBAFF in the presence or absence of inhibitors against JAK2 (AG490) and JAK3 (WHI-P131), and the production of IL-6 by the cells was measured by ELISA. The expression levels of the kinases were analyzed by quantitative PCR. Results Quantitative PCR showed that the expression level of BAFF-R was significantly elevated in pSS monocytes (2.1-fold) compared to normal monocytes. In accordance with these data, FACS analysis indicated that approximately 60% of pSS monocytes were BAFF-R-positive while only about 25% of normal monocytes were positive. These data suggest that the increased expression of BAFF-R in pSS monocytes was attributed to an increase in the population of BAFF-R-positive cells among monocytes of pSS patients. Notably, the expression of level of BAFF-R varied among pSS patients and was positively correlated with the IL-6 production by monocytes. We found that a JAK3 inhibitor showed strong suppressive effect on the production of IL-6 by the monocytes in a dose dependent manner. These data strongly suggest that sBAFF activates monocytes to induce IL-6 production through a JAK pathway. Interestingly, stimulation of pSS monocytes with sBAFF induced the expression of JAK3 while normal monocytes did not significantly respond to the stimulation. Conclusion Though the contribution of this induction to the overproduction of IL-6 remains obscure, we believe that these abnormalities as well as the up-regulation of BAFF-R underlie the abnormal production of IL-6 by pSS monocytes.

Increased microRNA-155 expression in the serum and peripheral monocytes in chronic HCV infection

BackgroundHepatitis C Virus (HCV), a single stranded RNA virus, affects millions of people worldwide and leads to chronic infection characterized by chronic inflammation in the liver and in peripheral immune cells. Chronic liver inflammation leads to progressive liver damage. MicroRNAs (miRNA) regulate inflammation (miR-155, -146a and -125b) as well as hepatocyte function (miR-122).MethodsHere we hypothesized that microRNAs are dysregulated in chronic HCV infection. We examined miRNAs in the circulation and in peripheral monocytes of patients with chronic HCV infection to evaluate if specific miRNA expression correlated with HCV infection.ResultsWe found that monocytes from chronic HCV infected treatment-naïve (cHCV) but not treatment responder patients showed increased expression of miR-155, a positive regulator of TNFα, and had increased TNFα production compared to monocytes of normal controls. After LPS stimulation, miR-155 levels were higher in monocytes from cHCV patients compared to controls. MiR-125b, which has negative regulatory effects on inflammation, was decreased in cHCV monocytes compared to controls. Stimulation of normal monocytes with TLR4 and TLR8 ligands or HCV core, NS3 and NS5 recombinant proteins induced a robust increase in both miR-155 expression and TNFα production identifying potential mechanisms for in vivo induction of miR-155. Furthermore, we found increased serum miR-155 levels in HCV patients compared to controls. Serum miR-125b and miR-146a levels were also increased in HCV patients. Serum levels of miR-122 were elevated in cHCV patients and correlated with increased ALT and AST levels and serum miR-155 levels.ConclusionIn conclusion, our novel data demonstrate that miR-155, a positive regulator of inflammation, is upregulated both in monocytes and in the serum of patients with chronic HCV infection. Our study suggests that HCV core, NS3, and NS5 proteins or TLR4 and TLR8 ligands can mediate increased miR-155 and TNFα production in chronic HCV infection. The positive correlation between serum miR-155 and miR-122 increase in cHCV may be an indicator of inflammation-induced hepatocyte damage.

Differential Effect of Telmisartan and Amlodipine on Monocyte Chemoattractant Protein-1 and Peroxisome Proliferator-Activated Receptor-Gamma Gene Expression in Peripheral Monocytes in Patients With Essential Hypertension

Monocyte chemoattractant protein-1 (MCP-1) and peroxisome proliferator-activated receptor-γ (PPAR-γ) play a significant role in monocyte activation, vascular inflammation, and atherogenesis. Angiotensin receptor blockers and calcium channel blockers are antihypertensive drugs with established efficacy and a favorable safety profile. We investigated the effect of telmisartan--an angiotensin receptor blocker with PPAR-γ agonist activity--and amlodipine on the activation state of peripheral blood monocytes with respect to MCP-1 and PPAR-γ gene expression in hypertensives. We recruited 31 previously untreated patients with essential hypertension who were randomly assigned to receive treatment with telmisartan (n = 16) or amlodipine (n = 15). Blood samples were taken before and 3 months after therapy initiation. Mononuclear cells were isolated and mRNAs of MCP-1 and PPAR-γ were estimated by real-time quantitative reverse transcription-polymerase chain reaction each time. The 2 treatments decreased all blood pressure components significantly (p <0.001). In contrast, in the amlodipine group, MCP-1 gene expression was significantly downregulated after treatment with telmisartan (from 21.4 ± 20.5 to 8.1 ± 6.5, p = 0.009), whereas the amlodipine group did not show any significant change (12.5 ± 8.5 vs 17.6 ± 16.4, p = NS). In addition, PPAR-γ mRNA levels showed a significant increase in telmisartan-treated patients (from 20 ± 18.5 to 42.6 ± 36, p = 0.006) and no significant alterations in the amlodipine group (from 29.6 ± 42.5 to 24.2 ± 27.7, p = NS). In conclusion, treatment with telmisartan results in a significant attenuation of MCP-1 gene expression and an increase of PPAR-γ gene expression in peripheral monocytes in patients with essential hypertension. Our findings may provide new insights into the cardiovascular protection of telmisartan in hypertensives.

Positive correlation between pregnancy-associated plasma protein-A level and OX40 ligand expression in patients with acute coronary syndromes

Objective Increasing evidence show that serum pregnancy-associated plasma protein-A (PAPP-A) and OX40 ligand (OX40L) expression have been implicated in acute coronary syndromes (ACS). We investigated the relationship between PAPP-A level and OX40L in serum and membrane-bound OX40L in patients with ACS. Methods The present study included normal controls ( n = 30), patients with stable angina (SA) ( n = 60) and patients with ACS, including unstable angina (UA) ( n = 50) and acute myocardial infarction (AMI) ( n = 30). Serum concentrations of PAPP-A and soluble OX40L (sOX40L) were determined with Elisa, whereas the expression of OX40L on monocytes were analyzed with flow cytometry. Results The expression of OX40L in peripheral monocytes in patients with UA [25.6 ± 5.5 mean fluorescence intensity (MFI)] and AMI (29.4 ± 6.3 MFI) were significantly higher than those in patients with SA (10.6 ± 2.8 MFI) and controls (11.1 ± 3.5 MFI). Both sOX40L and PAPP-A in patients with UA (15.7 ± 4.9 ng/mL, 25.4 ± 6.8 μg/mL, respectively) and AMI (17.1 ± 5.3 ng/mL, 26.3 ± 5.6 μg/mL, respectively) were significantly higher than those in patients with SA (3.4 ± 1.4 ng/mL, 9.6 ± 2.1 μg/mL, respectively) and controls (3.9 ± 1.3 ng/mL, 8.5 ± 2.8 μg/mL, respectively) ( P < 0.001). Interestingly, a positive correlation was found between sOX40L, membrane-bound OX40L and serum PAPP-A levels (r 1 = 0.54, r 2 = 0.51; P < 0.0001). Both serum and membrane-bound OX40L and PAPP-A levels significantly correlated with complex coronary stenosis (r 1 = 0.56, r 2 = 0.55, r 3 = 0.40; P < 0.001). Conclusion PAPP-A level was significantly related to soluble and membrane-bound OX40L in patients with ACS. Enhanced level of serum PAPP-A and sOX40L might represent a prognostic marker for coronary disease activity.

DIFFERENTIAL EFFECT OF TELMISARTAN AND AMLODIPINE ON MONOCYTE CHEMOATRACTANT PROTEIN-1 AND PEROXISOME PROLIFERATOR ACTIVATED RECEPTORS GAMMA GENE EXPRESSION IN HYPERTENSIVES: PP.13.482

Objective: Monocyte chemoattractant protein-1 (MCP-1), and peroxisome proliferator-activated receptors gamma (PPAR-gamma) play a significant role in monocytes activation, vascular inflammation and atherogenesis. Angiotensin receptor blockers and calcium blockers are both antihypertensive drugs with established efficacy and favorable safety profile.We investigated the effect of telmisartan – angiotensin II type 1 receptor blocker with PPAR-gamma agonist activity – and amlodipine on activation state of peripheral blood monocytes with respect to MCP-1, and PPAR-gamma mRNA expression in patients with essential hypertension. Design and Method: We recruited 28 previously untreated patients with mild or moderate essential hypertension (16 male, aged 61 ± 10 years) who were randomly assigned to receive treatment with either telmisartan (n = 14) or amlodipine (n = 14), achieving a mean blood pressure below 140/80 mmHg. Blood samples were taken before and 3 months after therapy initiation. Mononuclear cells were isolated using anti-CD14 + antibodies and mRNAs were estimated by real-time quantitative reverse transcription-PCR and expressed as fold induction. Results: There were no significant changes in blood pressure (BP), glucose, and lipid parameters between baseline and 12 weeks after randomization in either group. Both treatments reduced all blood pressure components significantly (p < 0.001). In contrast with amlodipine group, MCP-1 gene expression was significantly downregulated after treatment with telmisartan (fold induction in telmisartan group 0.8 ± 0.35 versus 1.87 ± 0.9 in the amlodipine group, p = 0.03). Additionally, PPAR-gamma gene expression showed a significant increase in telmisartan treated patients (fold induction in telmisartan group 2.8 ± 4.4 versus 5.3 ± 2.9 in the amlodipine group, p = 0.04). Conclusions: Treatment with telmisartan results in a significant induction of PPAR-gamma gene expression and attenuation of MCP-1 gene expression in peripheral monocytes in patients with essential hypertension. Our findings may provide new insights into the benefits and cardiovascular protection of telmisartan in hypertensive patients.