Peripheral Blood Monocytes Of Patients Research Articles

HomeoboxC6 affects the apoptosis of human vascular endothelial cells and is involved in atherosclerosis.

Apoptosis of vascular endothelial cells (VECs) is highly important in the occurrence and development of atherosclerosis (AS). HomeboxC6 (HOXC6) is expressed in higher levels in multiple malignant tissues, and it influences the malignant biological behavior of the cancer cells. However, the effects of HOXC6 on AS and the apoptosis of VECs have not been fully elucidated. In this study, we demonstrated that HOXC6 expression was increased in aortic wall of AS rats and peripheral blood monocytes of patients with coronary heart disease. Furthermore, it was uncovered that BAX expression was upregulated, while BCL-2 expression was downregulated in the aortic wall of AS rats. The apoptosis of human VECs (HVECs) cultured normally or treated with oxidized low-density lipoprotein in vitro was decreased after transfection with HOXC6-siRNA. Moreover, the results of Western blot analysis unveiled that the expressions of proapoptotic proteins, such as BAX, caspase-3, cleaved-caspase-3, and caspase-9 were reduced, while the expression of antiapoptotic protein, BCL-2, was elevated. Meanwhile, mRNA and protein expressions of phospholipase C beta (PLCβ) were decreased, the phosphorylation levels of protein kinase C zeta (PKCζ) and nuclear transcription factor-κB-p65 (NF-κBp65) and the membrane translocation of PKCζ were reduced as well. Besides, the expression of interleukin-18 (IL-18) protein was downregulated. However, after overexpression of HOXC6, the opposite trends of the abovementioned indices were observed. Furthermore, the inhibition of apoptosis induced by HOXC6-siRNA was reversed by lysophosphatidylcholine, an activator of PKCζ. Taken together, our results indicated that HOXC6 can promote the apoptosis of HVECs and may be involved in the occurrence and development of AS, which may be partially associated with the activation of PLCβ/PKCζ/NF-κBp65/IL-18 signaling pathway.

Functional activity of peripheral blood monocytes in patients with opisthorchiasis coupled to liver fibrosis intensity

The aim is to examine activity of peripheral blood monocytes from patients with opisthorchiasis coupled to intensity of liver fibrosis. Materials and Methods. A total of 74 patients with chronic opisthorchiasis (39 males and 35 females, average age 42.3 years) and 32 apparently healthy subjects (17 males and 15 females, average age 41.5 years) aged 24 to 60 years were enrolled. Opisthorchiasis was mainly diagnosed by parasitological assays to detect adult parasite eggs or bodies in duodenal contents and/or feces in all 74 patients. Liver fibrosis was assessed by elastometry according to the METAVIR scale. Functional activity of peripheral blood monocytes was performed in all patients with opisthorchiasis and healthy individuals from the control group by chemiluminescent analysis to measure intensity of reactive oxygen species production in lucigenin- and luminol-dependent spontaneous and zymosan-induced reactions. Results. Liver fibrosis F2 and F3—F4 stage according to METAVIR scale was found in 20.3% and 17.6% of patients with opisthorchiasis, respectively. While analyzing total pool of reactive oxygen species in the luminol-dependent process in patients with opisthorchiasis, a significantly decreased monocyte functional activity was observed as compared to healthy subjects that was evidenced by significantly decreased maximum intensity of produced reactive oxygen species as well as area under the chemiluminescence curve both in spontaneous and zymosan-induced reaction. Such parameters in liver fibrosis F3—F4 compared to F0—F1 in zymosan-induced response were lowered. Monocyte functional activity in spontaneous luminol-dependent reaction did not differ significantly depending on liver fibrosis intensity in patients with opisthorchiasis. The phagocytosis activation index in patients with opisthorchiasis with liver fibrosis F3—F4 compared to F0—F1 and F2 stage was lower. Similar changes were observed in the lucigenin-dependent reaction. Conclusion. The data obtained undoubtedly provide promising evidence to interpret the mechanisms behind liver fibrosis in patients with opisthorchiasis and create new opportunities for development of diagnostic and therapeutic technologies.

Effect of Yangxinshi tablets on the phenotype and function of monocytes in patients with unstable angina pectoris of coronary heart disease

Objective: The objective of this study was to observe the effect of Yangxinshi tablets on the phenotype and function of peripheral blood monocytes in patients with unstable angina of coronary heart disease (CHD) and to explore the possible mechanism of Yangxinshi tablets in the treatment of CHD. Methods: A total of 100 patients with unstable angina of CHD were randomly divided into two groups: Group A – treatment group and Group B – control group. The phenotypic and functional changes in blood monocytes and the changes in serum inflammatory factors before and after treatment were compared in the two groups. Results: The expression of CD14+CD163+ interleukin (IL)-10+ was significantly higher in the control group than in the patients with unstable angina of CHD, whereas the expression of CD14+CD163−IL-12+ was lower. The concentration of IL-10 was higher in the control group than in the patients with unstable angina, whereas the concentration of tumor necrosis factor-α (TNF-α) and IL-12 was significantly lower. In Group A, the expression of CD14+CD163+, CD14+CD206+, and CD14+CD163+CD206+ in peripheral blood monocytes increased after treatment than before treatment, whereas the expression of CD14+CD163−CD206− decreased. The expression of CD14+CD163−IL-12+ decreased after treatment than before, whereas CD14+CD163+IL-10+ expression increased. The serum concentration of IL-10 in Group A was higher after treatment than before, whereas that of IL-12 and TNF-α was lower. In Group B, the phenotype and function of the peripheral blood monocytes remained unchanged. Conclusion: Yangxinshi tablet therapy can change the phenotype of the peripheral blood monocytes in patients with unstable angina of CHD. Yangxinshi tablet therapy changes the inflammatory state in patients with an increase in the expression of anti-inflammatory factors and a decrease in the expression of inflammatory factors.

TLR expression on peripheral blood monocytes in patients with psoriasis

Toll-like receptors (TLRs) are signalling pattern-recognition receptors, which play an important part in initiating the immune response in psoriasis. The available literature has little information about study of these receptors in blood. The purpose of the present work was to study the level of expression of TLR2 and TLR4 types on blood monocytes in a psoriasis patient. Within 2016-2018, TLR2 and TLR4 were examined thrice in the blood of a patient with psoriasis during its exacerbation before the beginning of his treatment. The expression of surface receptors CD282 (TLR2) and CD284 (TLR4) on peripheral blood monocytes was studied by the method of flow cytometry with use of monoclonal antibodies. The obtained data show that the expression of TLR2 on peripheral blood monocytes was high, while that of TLR4 was mostly within its reference values of 3.3±0.1 mfi. Hence, the conducted studies have shown that the expression of TLR2 on peripheral blood monocytes in all three studies was high, thereby demonstrating involvement of this factor into the pathogenesis of the above disease. During all three blood examinations, monocytes revealed an extremely high intensity of TLR2 fluorescence on studied cells that exceeded the same intensity in healthy subjects by 1.5-2 times. The expression of TLR4 on peripheral blood monocytes during all three studies was within its reference values, this fact demonstrating absence of any information value of the above factor in the pathogenesis of psoriasis.

AP-1 (Activated Protein-1) Transcription Factor JunD Regulates Ischemia/Reperfusion Brain Damage via IL-1β (Interleukin-1β).

Background and Purpose- Inflammation is a major pathogenic component of ischemia/reperfusion brain injury, and as such, interventions aimed at inhibiting inflammatory mediators promise to be effective strategies in stroke therapy. JunD-a member of the AP-1 (activated protein-1) family of transcription factors-was recently shown to regulate inflammation by targeting IL (interleukin)-1β synthesis and macrophage activation. The purpose of the present study was to assess the role of JunD in ischemia/reperfusion-induced brain injury. Methods- WT (wild type) mice randomly treated with either JunD or scramble (control) siRNA were subjected to 45 minutes of transient middle cerebral artery occlusion followed by 24 hours of reperfusion. Stroke size, neurological deficit, plasma/brain cytokines, and oxidative stress determined by 4-hydroxynonenal immunofluorescence staining were evaluated 24 hours after reperfusion. Additionally, the role of IL-1β was investigated by treating JunD siRNA mice with an anti-IL-1β monoclonal antibody on reperfusion. Finally, JunD expression was assessed in peripheral blood monocytes isolated from patients with acute ischemic stroke. Results- In vivo JunD knockdown resulted in increased stroke size, reduced neurological function, and increased systemic inflammation, as confirmed by higher neutrophil count and lymphopenia. Brain tissue IL-1β levels were augmented in JunD siRNA mice as compared with scramble siRNA, whereas no difference was detected in IL-6, TNF-α (tumor necrosis factor-α), and 4-hydroxynonenal levels. The deleterious effects of silencing of JunD were rescued by treating mice with an anti-IL-1β antibody. In addition, JunD expression was decreased in peripheral blood monocytes of patients with acute ischemic stroke at 6 and 24 hours after onset of stroke symptoms compared with sex- and age-matched healthy controls. Conclusions- JunD blunts ischemia/reperfusion-induced brain injury via suppression of IL-1β.

Sodium Chloride Aggravates Arthritis via Th17 Polarization.

PurposeSodium chloride (NaCl) has been proposed as a driving factor in autoimmune diseases through the induction of pathogenic CD4+ T helper cells that produce interleukin-17 (Th17 cells). This study investigated the effects of NaCl on inflammatory arthritis in mice and humans.Materials and MethodsCollagen-induced arthritis (CIA) mice were fed a normal or high-salt diet ad libitum, and clinical and histologic features of arthritis were evaluated. The proportion of Th17 cells in the spleens of CIA mice fed a normal or high-salt diet was evaluated by flow cytometry, and the expression of IL-17 in joints and intestines was determined by immunohistochemical staining. We also analyzed the effect of NaCl on Th17 differentiation from peripheral blood monocytes of patients with rheumatoid arthritis (RA) and osteoarthritis (OA) and evaluated the contents of sodium and IL-17 in the synovial fluid of RA and OA patients.ResultsNaCl increased murine and human Th17 cell differentiation in a dose-dependent manner. Clinical and histological arthritis was more severe in the high-salt-fed CIA mice, compared to control CIA mice. The proportion of Th17 cells among splenocytes was higher in CIA mice fed a high-salt diet. Expression of synovial and intestinal IL-17 was also higher in high-salt-fed CIA mice. Comparison of synovial fluid between RA patients and OA patients revealed that Na+ and IL-17 were more abundant in RA synovial fluid.ConclusionThis study suggests that NaCl can aggravate arthritis by affecting Th17 differentiation. Accordingly, limiting salt intake may be helpful for treating inflammatory arthritis, such as RA.

Signature miRNAs in peripheral blood monocytes of patients with gastric or breast cancers.

The dysregulation of microRNAs (miRNAs), key posttranscriptional regulators of gene expression, is closely associated with cancer development. However, the miRNAs of monocytes, important cells of tumour immunity, have not been extensively explored. In the present study, the differentially expressed miRNAs of blood monocytes derived from gastric and breast cancer patients and healthy donors were characterized. The results indicated that 74 miRNAs were upregulated and 46 miRNAs were downregulated in monocytes of patients with breast or gastric cancers compared with the healthy donors, suggesting that these 120 miRNAs from transformed monocytes were associated with cancers. The differentially expressed miRNAs, 38 of which were novel, were further validated using quantitative real-time PCR. As an example, the results showed that miR-150-5p downregulated the CCR2 expression in monocytes by targeting Notch 3, thus leading to the suppression of tumorigenesis. The target gene analysis showed that 36 of the 120 miRNAs targeted cancer-related genes. KEGG pathway analysis indicated that the cancer-associated miRNAs were involved in pathways related to cancers, such as the HIF-1 signalling and the mTOR signalling pathways. Thus, our study provided new clues to comprehensively understand the relationship between miRNAs and cancers.

Sirtuin 5 as a novel target to blunt blood–brain barrier damage induced by cerebral ischemia/reperfusion injury

BackgroundIn acute ischemic stroke (AIS) patients, impaired blood–brain barrier (BBB) integrity is associated with hemorrhagic transformation and worsened outcome. Yet, the mechanisms underlying these relationships are poorly understood and consequently therapeutic strategies are lacking. This study sought to determine whether SIRT5 contributes to BBB damage following I/R brain injury. Methods and resultsSIRT5 knockout (SIRT5−/−) and wild type (WT) mice underwent transient middle cerebral artery (MCA) occlusion (tMCAO) followed by 48h of reperfusion. Genetic deletion of SIRT5 decreased infarct size, improved neurological function and blunted systemic inflammation following stroke. Similar effects were also achieved by in vivo SIRT5 silencing. Immunohistochemical analysis revealed decreased BBB leakage and degradation of the tight junction protein occludin in SIRT5−/− mice exposed to tMCAO as compared to WT. In primary human brain microvascular endothelial cells (HBMVECs) exposed to hypoxia/reoxygenation (H/R), SIRT5 silencing decreased endothelial permeability and upregulated occludin and claudin-5; this effect was prevented by the PI3K inhibitor wortmannin. Lastly, SIRT5 gene expression was increased in peripheral blood monocytes (PBMCs) of AIS patients at 6h after onset of stroke compared to sex- and age-matched healthy controls. ConclusionSIRT5 is upregulated in PBMCs of AIS patients and in the MCA of WT mice exposed to tMCAO; SIRT5 mediates I/R-induced brain damage by increasing BBB permeability through degradation of occludin. This effect was reproduced in HBMVECs exposed to H/R, mediated by the PI3K/Akt pathway. Our findings shed new light on the mechanisms of I/R-dependent brain damage and suggest SIRT5 as a novel therapeutic target.

Иммунофенотипическая характеристика моноцитов периферической крови пациентов с болезнью Альцгеймера и синдромом мягкого когнитивного снижения

Aim : to study the features of subpopulation composition of peripheral blood monocytes in patients with Alzheimer's disease (AD) and with amnesic mild cognitive impairment syndrome (aMCI). Material and methods : 20 AD patients aged 73.8 ± 6.8 years, and 10 patients with aMCI aged 61.8 ± 9.6 years enrolled in the study were examined. 16 mentally and somatically healthy age- and gender-matched persons were examined as controls. The peripheral venous blood of patients and controls was the material for the study. The relative number of monocytes was determined with the help of FC-500 Flow Суtofluorometer [Beckman Coulter, USA] using the same settings for all samples. Results : the study revealed a significant decrease of the relative number of classical subpopulations (p < 0.01) and an increase of the proinflammatory monocytes level (p < 0.01) in the group of AD patients compared with the control group. The severity of changes of the monocytes subpopulation composition depended on the dementia severity. The sharpest change of the monocytes composition compared to controls was typical for patients with moderate dementia (p < 0.001). An increase of the subpopulation level of proinflammatory monocytes was also revealed in patients with aMCI (p < 0.05). Conclusion : the study results have revealed the redistribution of monocyte subpopulations in AD and aMCI patients: the decrease in the classical monocytes subpopulation on the background of the increase of the monocytes subpopulation with proinflammatory activity.

Impairment of migration related molecule expression on peripheral blood monocytes in patients with hereditary diffuse leukoencephalopathy with spheroids

Impairment of migration related molecule expression on peripheral blood monocytes in patients with hereditary diffuse leukoencephalopathy with spheroids

Ultrasound-guided percutaneous portal transplantation of peripheral blood monocytes in patients with liver cirrhosis.

Background/AimsLiver transplantation offers the only definite cure for cirrhosis but lacking donors is problem. Stem cell therapy is attractive in this setting. In this study, we aimed to explore the safety and efficacy of ultrasound-guided percutaneous portal transplantation of peripheral blood monocyte cell (PBMC) in cirrhotic patients.MethodsA total of nine decompensated cirrhotic patients were randomized into three groups: group 1 (n = 3) was control group, group 2 (n = 3) received granulocyte-colony stimulating factor (G-CSF) mobilization for 3 days, and group 3 (n = 3) received G-CSF mobilized PBMCs by leukapheresis and PBMC transplantation through ultrasound-guided percutaneous portal vein puncture. Liver function and clinical features were evaluated.ResultsAt baseline, the Child-Turcotte-Pugh and the model for end-stage liver disease scores were comparable in study groups. Compared with group 1, there was a tendency to improve liver function in group 3 at 6 months after treatment. Treatment was tolerable and no complications were encountered related to the G-CSF mobilization or percutaneous portal administration of PBMCs. Imaging studies showed patent portal veins at the end of the study period.ConclusionsAutologous PBMC transplantation through ultrasound-guided percutaneous portal vein puncture could be considered as a safe alternative treatment for decompensated cirrhotic patients.

The cancer vaccine therapy using DCs derived from iPS cells (iPSDCs) expressing TAA gene.

768 Background: Dendritic cells (DCs) are potent antigen-presenting cells (APCs) that play a critical role in the initiation of anti-tumor immune responses. Many cancer patients have previously been treated by the cancer vaccine therapy using DCs worldwide. We have employed a study of a cancer vaccine therapy using genetically modified DCs expressing tumor-associated antigen (TAA) gene. Clinically DCs are generated from the peripheral blood monocytes of patients. Thus the number of monocytes and potential of them are limited, so they are serious obstacle. Recent studies have revealed that induced pluripotent stem (iPS) cells can be generated from murine fibroblasts. Furthermore, it has been reported that DCs can be successfully derived from murine iPS cells (iPSDCs). If the therapeutic efficacy of iPSDCs is equivalent to that of naive DCs, then the above-mentioned problems may be solved. Methods: We have induced iPSDCs from murine iPS cells by 4 steps and examined the efficacy as APCs of iPSDCs compared with naive DCs. We also examined whether a vaccine therapy using genetically modified iPSDCs can induce strong therapeutic antitumor immunity compared with naive DCs. Next, we examined the therapeutic antitumor immunity of iPSDCs expressing CEA gene compared with that of naive DCs in pre-clinical study with CEA transgenic mice. Results: We have clarified that genetically modified iPSDCs have an equal efficacy as APCs and TAA-specific therapeutic antitumor immunity, equivalent to naive DCs. And we also have clarified that genetically modified iPSDCs expressing CEA gene have a TAA-specific therapeutic antitumor immunity. Conclusions: This vaccine strategy using genetically modified iPSDCs has an equal capacity with naive DCs in terms of a therapeutic efficacy. Now, we are engaging another pre-clinical study with human, in an effort to apply in a clinical setting.

Decreased Histone Deacetylase 2 (HDAC2) in Peripheral Blood Monocytes (PBMCs) of COPD Patients.

BackgroundHistone deacetylase 2 (HDAC2) is a class I histone deacetylase family member that plays a critical role in suppressing inflammatory gene expression in the airways, lung parenchyma, and alveolar macrophages in patients with chronic obstructive pulmonary disease (COPD). However, the expression of HDAC2 in peripheral blood monocytes (PBMCs), nuclear factor kappa B (NF-κB) p65, and serum inflammatory cytokine levels in COPD patients, smokers, and non-smokers remains unclear.MethodsPBMCs were obtained from COPD patients, healthy smokers, and healthy nonsmokers. The HDAC2 and NF-κB p65 expression were quantified by Western Blot. HDAC activity was assessed by an HDAC fluorometric immunoprecipitation activity assay kit. Serum tumor necrosis factor-alpha (TNF-α) and interleukin-8 (IL-8) levels were measured by ELISA.ResultsHDAC2 expression and HDAC activity were decreased in PBMCs in COPD patients compared with smokers and non-smokers. Increased NF-κB p65 expression, serum TNF-α and IL-8 levels were observed in COPD patients compared with nonsmokers. The FEV1%pred was positively correlated with HDAC2 expression and HDAC activity in COPD patients. Smokers had decreased HDAC activity, increased NF-κB p65 expression and serum TNF-α compared with nonsmokers.ConclusionsHDAC2 expression was decreased in PBMCs of COPD patients and was correlated with disease severity. The reduction of HDAC2 expression not only directly enhances the expression of inflammatory genes, but may account for the activation of NF-κB mediated inflammation. Decreased HDAC2 may serve as a potential biomarker of COPD and predict the decline of lung function.

Expression of TLR2 and TLR4 on peripheral blood monocytes during exacerbation of atopic dermatitis

In atopic dermatitis (AD), monocytes, which accumulate in the inflamed skin, are characterized by a significantly impaired Toll-like receptors (TLR) expression and TLR2-mediated cytokine secretion. However, data on expression of TLR on monocytes of peripheral blood (PB) in AD are not available. To investigate TLR2 and TLR4 expression on PB monocytes during AD exacerbation and to assess the relationships between TLR expressions with AD clinical severity and with serum interleukin (IL) 4, IL-10, and IL-17a levels. The objective Scoring Atopic Dermatitis index, TLR2 and TLR4 expression on CD14(+) human leukocyte antigen-DR (HLA-DR(+)) PB monocytes by flow cytometry, serum IL-4, IL-10, IL-17a (enzyme-linked immunosorbent assay) and total immunoglobulin E levels were measured at study entry and after 4 months in patients with AD and healthy controls. Eighty-two patients with AD, 35 women (45.1%) and 47 men (54.9%), mean (standard deviation [SD]) age, 42.2 ± 11.5 years, were included. Thirty healthy volunteers served as controls. We observed a significant difference in the levels of TLR2 expression in the CD14(+) HLA-DR(+) PB monocytes of patients with AD (mean [SD], 51.6 ± 23.1% and 264 ± 118 cells/mm(3)) at exacerbation (but not at the end of the 4-month postexacerbation period) compared with the healthy control subjects (mean [SD], 22.3 ± 10.6% and 105 ± 50 cells/mm(3); p < 0.001). TLR4 expression in PB monocytes was significantly greater in AD (mean [SD], 50.1 ± 20.9% and 275 ± 114 cells/mm(3)) than in the healthy subjects (mean [SD], 31.2 ± 8.7% and 147 ± 41 cells/mm(3); p < 0.001) both at exacerbation and at the 4-month postexacerbation period. Significant correlations between TLR2(+) (but not TLR4(+)) PB monocytes and the objective Scoring Atopic Dermatitis index (r = 0.604, p < 0.001), serum levels of IL-17a and TLR2(+) PB monocytes (r = 0.416, p = 0.027), and IL-4 and TLR2(+) PB monocytes (r = -0.307, p = 0.014) were observed during AD exacerbation. PB CD14(+) HLA-DR(+) TLR2(+) monocytes might have a role in the skewing of a T-helper 2/T-helper 17-mediated immune response during AD flare.

Heparin-treated dendritic cells promote Th0 to Th1 differentiation via the Toll-like receptor 3 in peripheral blood monocytes of patients with chronic hepatitis B

To investigate the mechanisms underlying the ability ofheparin-treated dendritic cells (DCs) to promote Th0 to Th1 differentiation in chronic hepatitis B (CHB). Peripheral blood mononuclear cells (PBMCs) were isolated from CHB patients and cultured in RPMI-1640 with recombinant GM-CSF and IL-4 with or without heparin to obtain DCs for study. The levels of Toll-like receptors (TLRs) on the DCs were measured using FACS and qPCR techniques.DC subsets with high expression of TLRs were selected for analysis of functional changes by treatment with the corresponding TLR-siRNA. The CD4+ T cell subpopulation was purified from peripheral blood by Dynal immunomagnetic beads, and then the production of IL-12 by DCs in the presence of poly(I:C) or R848 and ofIFN and IL-4 by Th cells co-cultured with DCs was evaluated by ELISA. The t-test was used for statistical analysis. TLR3 expression, and not expression of TLR 7 or TLR8,was significantly increased in heparin-treated DCs as compared to levels detected in the DCs without heparin treatment (t =2.849,P less than 0.05;t =3.027,P less than 0.05). The level of IL-12 produced by heparin-treated DCs stimulated with poly(I:C) was obviously higher than that produced by DCs without heparin treatment and stimulated with poly(I: C) (t =8.68,P less than 0.01) or with R848 (t =19.01,P less than 0.01). However, the IL-12 production by TLR3-siRNA transfected-DCs was significantly reduced (t =31.49, P less than 0.01).When Th cells from allogenic patients with CHB were co-cultured with the TLR3-siRNA transfectedDCs, the frequency ofCD4+ IFN+ cells was significantly reduced (1.64+/-0.57% vs.6.31+/-0.88%,P less than 0.01),as was the capability of Thl to generate IFNg (t =20.83,Pless than 0.01). Heparin may have up-regulated the TLR3 expression level of DCs, and sequentially promoted Th0 to Th1 differentiation.

Monocytes from patients with rheumatoid arthritis and type 2 diabetes mellitus display an increased production of interleukin (IL)-1β via the nucleotide-binding domain and leucine-rich repeat containing family pyrin 3(NLRP3)-inflammasome activation: a possible implication for therapeutic decision in these patients.

A better understanding about the mechanisms involved in the pathogenesis of type 2 diabetes mellitus (T2D) showed that inflammatory cytokines such as tumour necrosis factor (TNF) and interleukin (IL)-1β play a pivotal role, mirroring data largely reported in rheumatoid arthritis (RA). IL-1β is produced mainly by monocytes (MO), and hyperglycaemia may be able to modulate, in the cytoplasm of these cells, the assembly of a nucleotide-binding domain and leucine-rich repeat containing family pyrin (NLRP3)-inflammosome, a cytosolic multi-protein platform where the inactive pro-IL-1β is cleaved into active form, via caspase-1 activity. In this paper, we evaluated the production of IL-1 β and TNF, in peripheral blood MO of patients affected by RA or T2D or both diseases, in order to understand if an alteration of the glucose metabolism may influence their proinflammatory status. Our data showed, after 24 h of incubation with different glucose concentrations, a significantly increased production of IL-1β and TNF in all evaluated groups when compared with healthy controls. However, a significant increase of IL-1β secretion by T2D/RA was observed when compared with other groups. The analysis of relative mRNA expression confirmed these data. After 24 h of incubation with different concentrations of glucose, our results showed a significant increase in NLRP3 expression. In this work, an increased production of IL-1β by MO obtained from patients affected by both RA and T2D via NLRP3-inflammasome activation may suggest a potential IL-1β targeted therapy in these patients.

Bifidobacteria Enhance Antigen Sampling and Processing by Dendritic Cells in Pediatric Inflammatory Bowel Disease.

Bifidobacteria have been reported to reduce inflammation and contribute to intestinal homeostasis. However, the interaction between these bacteria and the gut immune system remains largely unknown. Because of the central role played by dendritic cells (DCs) in immune responses, we examined in vitro the effects of a Bifidobacteria mixture (probiotic) on DC functionality from children with inflammatory bowel disease. DCs obtained from peripheral blood monocytes of patients with Crohn's disease (CD), ulcerative colitis, and noninflammatory bowel disease controls (HC) were incubated with fluorochrome-conjugated particles of Escherichia coli or DQ-Ovalbumin (DQ-OVA) after a pretreatment with the probiotic, to evaluate DC phenotype, antigen sampling and processing. Moreover, cell supernatants were collected to measure tumor necrosis factor alpha, interferon gamma, interleukin 17, and interleukin 10 production by enzyme-linked immunosorbent assay. DCs from CD children showed a higher bacteria particles uptake and DQ-OVA processing after incubation with the probiotic; in contrast, DC from both ulcerative colitis and HC showed no significant changes. Moreover, a marked tumor necrosis factor alpha release was observed in DC from CD after exposure to E. coli particles, whereas the probiotic did not affect the production of this proinflammatory cytokine. In conclusion, the Bifidobacteria significantly improved the antigen uptake and processing by DCs from patients with CD, which are known to present an impaired autophagic functionality, whereas, in DCs from ulcerative colitis and HC, no prominent effect of probiotic mixture was observed. This improvement of antigen sampling and processing could partially solve the impairment of intestinal innate immunity and reduce uncontrolled microorganism growth in the intestine of children with inflammatory bowel disease.

Suppression of tumor growth by a M2b macrophage reverter in humanized murine chimeras created with peripheral blood monocytes of patients with advanced stage hepatocellular carcinoma (TUM6P.967)

Abstract Previously, we have reported that humanized murine chimeras created with peripheral blood monocytes from patients with early stage hepatocellular carcinoma (E-hccM) are resistant against tumor inoculation, while tumors grow in the same chimeras created with advanced stage patient monocytes (A-hccM). A majority of E-hccM are shown to be M1 monocytes, and a predominance of M2b monocytes is shown in A-hccM. In this study, we tried to suppress tumor growth in humanized chimeras created with A-hccM by CCL1 antisense ODN treatment, which reverts Mf from M2b to resident. Chimeras, created in X-irradiated NSG mice after inoculation of A-hccM (i.v., 1 x 106 cells/mouse, every 2 weeks), were inoculated with HepG2 cells (2 x 106 cells/mouse, s.c.). CCL1 antisense ODN (6 mg/mouse) were administered 4 times within 36 hrs of each A-hccM inoculation. In the results, 843 ± 227 mm3 of tumors were detected in the chimeras inoculated with A-hccM 8 weeks after the tumor inoculation, while tumors were not demonstrated in A-hccM chimeras treated with CCL1 antisense ODN, and 753 ± 165 mm3 of the tumors were detected in A-hccM chimeras treated with scrambled ODN. A-hccM were characterized as IL-12-IL-10+CCL1+iNOS-CD14+ cells, while IL-12+IL-10-CCL1-iNOS+CD14+ cells with tumoricidal activities were detected in A-hccM treated with the antisense ODN. These results indicate that M2b monocytes present in A-hccM play a key role in the tumor growth seen in advanced stage hepatocellular carcinoma patients.

Involvement of TLR2-MyD88 in abnormal expression of miR-146a in peripheral blood monocytes of patients with chronic hepatitis C.

miR-146a is an immunoregulatory microRNA closely associated with viral infection. This study investigated the expression changes of miR-146a in peripheral blood monocytes of HCV-infected patients and the mechanism by which the THP-1 cells were stimulated with HCV core protein in vitro. It was found that in the peripheral blood monocytes of HCV-infected patients, miR-146a expression was upregulated. After treated by interferon/ribavirin, miR-146a expression was decreased when HCV RNA became undetectable. HCV core could directly stimulate THP-1 cells to produce miR-146a. Silencing TLR2 and MyD88 could significantly inhibit the expression of miR-146a. It was concluded that the expression of miR-146a in peripheral blood monocytes of HCV-infected patients was abnormally increased. The TLR2-MyD88 signaling pathway may take part in the overexpression of miR-146a in monocytes stimulated with HCV core protein.

QUANTIFICATION AND PHENOTYPIC CHARACTERIZATION OF MONOCYTES SUBSETS IN HUMAN ATHEROSCLEROSIS

QUANTIFICATION AND PHENOTYPIC CHARACTERIZATION OF MONOCYTES SUBSETS IN HUMAN ATHEROSCLEROSIS