Elevated level of some chemokines in plasma of gastric cancer patients

Introduction : Gastric cancer is one of the most common cancer-related causes of death. This is mainly due to the lack of good noninvasive method/biomarkers suitable for early-tumour diagnosis and planning of further therapy modalities. Chemokines play an important role in cancer progression and metastasis formation. In gastric cancer patients, clinical relevance of CXCL12 and CCL5 level has been postulated. Aim of the study : Efforts were undertaken to examine whether expanded chemokine range may be relevant for evaluation of preoperative staging of gastric cancer patients. Material and methods : Plasma from 66 gastric cancer patients and 11 healthy controls was obtained, and CCL2, CCL3, CCL4, CCL5, CXCL8, CXCL9, and CXCL10 levels were determined by flow cytometry FlexSet system. Results : In gastric cancer patients’ plasma an increased level of CCL2, CCL4, CCL5, CXCL8, CXCL9, and CXCL10 was observed. In the case of CCL2, CXCL9, and CXCL10, the chemokine levels correlated with advanced (III and IV in TNM classification) disease stage. In the case of CCL4, CCL5, and CXCL8, elevated levels were observed in all cancer patients in comparison to healthy donors. Conclusions : The accuracy of preoperative diagnosis in gastric cancer may include the monitoring of a wide range of chemokines in patients’ plasma. Increased levels of chemokines may warn that the disease is more advanced than conventional diagnostic procedures suggest

Transition metal containing particulate matter promotes Th1 and Th17 inflammatory response by monocyte activation in organic and inorganic compounds dependent manner

In recent years, a significant increase in the frequency of disorders caused by air pollutants has been observed. Here we asked whether transition metal-containing particulate matter (TMCPM), a component of air pollution, has an effect on the activity of human CD4+ T cell subsets (Th1, Th2, Th17, and Treg). Peripheral blood mononuclear cells (PBMC) from healthy donors were cultured with or without NIST (SRM 1648a—standard urban particulate matter purchased from the National Institute for Standards and Technology) and LAP (SRM 1648a particulate matter treated within 120 min with cold oxygen plasma) preparations of TMCPM, differing in organic compounds content. Data show that TMCPM treatment increased the level of CD4+ cells positive for IFN-γ and IL-17A, specific for Th1 and Th17 cells, respectively. Moreover, a substantial decrease in frequency of Foxp3 positive CD4+ cells was observed in parallel. This effect was more pronounced for NIST particles, containing more organic components, including endotoxin (LPS - lipopolysaccharide) and required the presence of monocytes. Inactivation of LPS by treatment of TMCPM with polymyxin B reduced the inflammatory response of monocytes and Th subsets but did not abolish this activity, suggesting a role of their inorganic components. In conclusion, treatment of human PBMC with TMCPM skews the balance of Th1/Th2 and Treg/Th17 cells, promoting polarization of CD4+ T cells into Th1 and Th17 subsets. This phenomenon requires activation of monocytes and depends on the organic and inorganic fractions, including endotoxin content in TMCPM, as significantly higher inflammatory response was observed for the NIST comparing to LAP. This observation may shed a new light on the role of TMCPM in development and exacerbation of allergies, inflammatory, and autoimmune disorders

The role of CD44H molecule in the interactions between human monocytes and pancreatic adenocarcinoma-derived microvesicles

Introduction. CD44H is a transmembrane molecule important for cell-cell and cell-extracellular matrix interactions. In monocytes, CD44H is implicated in phagocytosis of particles coated by hyaluronan (HA). HA fragments were shown to induce chemokine secretion by monocytes. Tumour derived microvesicles (TMVs), which are small membrane fragments derived from tumour cells can carry fragments of HA. The aim of the study was to examine whether monocyte’s CD44H is involved in the engulfment of pancreatic adenocarcinoma-derived microvesicles and in the production of chemokines induced by TMVs. Materials and methods. TMVs engulfment and chemokines’ secretion stimulated with TMVs were determined in control human monocytes and cells incubated with anti-CD44H monoclonal antibody (mAb) by flow cytometry and ELISA, respectively. Phosphorylation of STAT3, transcription factor essential for chemokines’ production and CD44 signal transduction, was determined by Western blotting. Results. Blocking of CD44H by anti-CD44H mAb on monocytes decreased the engulfment of TMVs and thesecretion of CCL4 and CCL5, but had no effect on CCL2, CCL3 and CXCL8. STAT-3 phosphorylation inmonocytes incubated with TMVs after CD44 blocking was also reduced. Conclusion. The results suggest that tumour-derived microvesicles (TMVs) may carry bioactive cargo(s) which induces STAT3 dependent signalling pathway in human monocytes via CD44 molecules

T Lymphocyte-Derived Exosomes Transport MEK1/2 and ERK1/2 and Induce NOX4-Dependent Oxidative Stress in Cardiac Microvascular Endothelial Cells

Background: Activation of endothelial cells by inflammatory mediators secreted by CD4+ T lymphocytes plays a key role in the inflammatory response. Exosomes represent a specific class of signaling cues transporting a mixture of proteins, nucleic acids, and other biomolecules. So far, the impact of exosomes shed by T lymphocytes on cardiac endothelial cells remained unknown. Methods and results: Supernatants of CD4+ T cells activated with anti-CD3/CD28 beads were used to isolate exosomes by differential centrifugation. Activation of CD4+ T cells enhanced exosome production, and these exosomes (CD4-exosomes) induced oxidative stress in cardiac microvascular endothelial cells (cMVECs) without affecting their adhesive properties. Furthermore, CD4-exosome treatment aggravated the generation of mitochondrial reactive oxygen species (ROS), reduced nitric oxide (NO) levels, and enhanced the proliferation of cMVECs. These effects were reversed by adding the antioxidant apocynin. On the molecular level, CD4-exosomes increased NOX2, NOX4, ERK1/2, and MEK1/2 in cMVECs, and ERK1/2 and MEK1/2 proteins were found in CD4-exosomes. Inhibition of either MEK/ERK with U0126 or ERK with FR180204 successfully protected cMVECs from increased ROS levels and reduced NO bioavailability. Treatment with NOX1/4 inhibitor GKT136901 effectively blocked excessive ROS and superoxide production, reversed impaired NO levels, and reversed enhanced cMVEC proliferation triggered by CD4-exosomes. The siRNA-mediated silencing of Nox4 in cMVECs confirmed the key role of NOX4 in CD4-exosome-induced oxidative stress. To address the properties of exosomes under inflammatory conditions, we used the mouse model of CD4+ T cell-dependent experimental autoimmune myocarditis. In contrast to exosomes obtained from control hearts, exosomes obtained from inflamed hearts upregulated NOX2, NOX4, ERK1/2, MEK1/2, increased ROS and superoxide levels, and reduced NO bioavailability in treated cMVECs, and these changes were reversed by apocynin. Conclusion: Our results point to exosomes as a novel class of bioactive factors secreted by CD4+ T cells in immune response and represent potential important triggers of NOX4-dependent endothelial dysfunction. Neutralization of the prooxidative aspect of CD4-exosomes could open perspectives for the development of new therapeutic strategies in inflammatory cardiovascular diseases

Chemokine receptors and chemokine production by CD34+ stem cell-derived monocytes in response to cancer cells

Background: The chemokine-chemokine receptor (CR) network is involved in the regulation of cellular infiltration of tumours. Cancer cells and infiltrating macrophages produce a whole range of chemokines. This study explored the expression of some CR and chemokine production by cord blood stem cell-derived CD34+ monocytes and their novel CD14++CD16+ and CD14+CD16– subsets in response to tumour cells. Material and Methods: CR expression was determined by flow cytometry and their functional activity by migration to chemoattractants. Monocytes were cultured with tumour cells and the chemokine content was assessed in culture supernatants. Results: CD14++CD16+ monocytes exhibited increased expression of chemokine (C-C) receptor (CCR) 1, while CD14+CD16– of CCR2, chemokine (C-X-C) receptor (CXCR) 1, 2 and 4. The increased expression of CCR2 on CD14+CD16– monocytes was associated with their enhanced migration to monocyte chemoattractant protein–1 (CCL2), MCP-3 (CCL7), MCP-2 (CCL8) and MCP-4 (CCL13), while that of CXCR1 and 2 to interleukin 8 (CXCL8), and CXCR4 to stromal cell-derived factor-1 (CXCL12). Tumour cells induced production of macrophage inflammatory protein-1α (CCL3) MIP-1β and regulated on activation normal T-cells expressed and secreted (CCL5) but not CCL2 or CXCL8, monokine induced by gamma interferon (CXCL9), interferon gamma-induced protein 10 (CXCL10). Conclusion: The studied monocyte subsets, in comparison to those from blood, exhibit different expression of CRs and response to the stimuli that occur from tumour cells

Open study to evaluate etidronate influence on bone markers, inflammation and RANKL/OPG system in ankylosing spondylitis (AS) patients with active inflammation : effect of etidronate on number and metabolic activity of proinflammatory subpopulation of blood monocytes CD14+CD16+ in AS patients - a pilot study

Cel pracy: Ocena wpływu 3-miesięcznego cyklicznego leczenia etydronianem (bisfosfonianem) na markery kostne, wskaźniki zapalne i układ RANKL/osteoprotegeryna (RANKL/OPG) u pacjentów z aktywnym zesztywniającym zapaleniem stawów kręgosłupa (ZZSK). Ocena wpływu etydronianu na liczbę prozapalnych monocytów CD14+CD16+ i ich aktywność metaboliczną u pacjentów z aktywnym ZZSK. Materiał i metody: Etydronian podawano (w dwóch 14-dniowych cyklach) 25 mężczyznom spełniającym nowojorskie kryteria rozpoznania ZZSK, u których stwierdzono zwiększone stężenia wskaźników zapalnych (OB i/lub CRP). Przed podaniem leku i po kolejnych cyklach oznaczano osteokalcynę (OC, marker kościotworzenia), telopeptyd β-CTx (CTX, marker resorpcji), rozpuszczalny ligand receptora aktywującego czynnik jądrowy κB (receptor activator of nuclear factor κB ligand – sRANKL) i osteoprotegerynę (OPG). Wyniki: Po leczeniu etydronianem stwierdzono istotne statystycznie zmniejszenie się stężeń CTX, OC i CRP oraz nieistotne statystycznie zmniejszenie stężenia sRANKL, natomiast zwiększenie stężenia OPG. Nie stwierdzono korelacji pomiędzy markerami kostnymi a CRP, sRANKL i OPG, a także pomiędzy CRP a sRANKL i OPG. Wykazano dodatnią korelację pomiędzy wskaźnikiem sRANKL/OPG wyjściowym i po 1. cyklu oraz wyjściowym i po 2. cyklu. Nie stwierdzono różnic w bezwzględnej liczbie subpopulacji monocytów, tj. prozapalnej (CD14+CD16+) i klasycznej (CD14++CD16–) przed 1. cyklem i po 1. cyklu etydronianu. Po stymulacji PHA uzyskano średnio ok. 80-krotny wzrost wydzielania immunosupresyjnej cytokiny IL-10 i ok. 40-krotny wzrost wydzielania TNF-α przez izolowane komórki jednojądrowe krwi obwodowej po 1. cyklu etydronianu. Wnioski: 1. Etydronian wykazuje działanie antyresorpcyjne i przeciwzapalne u mężczyzn chorych na ZZSK. 2. Nie stwierdzono korelacji pomiędzy CRP a markerami kostnymi, sRANKL, OPG i sRANKL/OPG, co sugeruje brak sprzężenia procesu zapalenia z remodelacją tkanki kostnej w ZZSK. 3. Etydronian nie wpływa na liczbę prozapalnych monocytów CD14+CD16+ we krwi obwodowej oraz zwiększa stymulowaną PHA syntezę cytokin przez jednojądrowe komórki krwi obwodowej, z przewagą wydzielania immunosupresyjnej IL-10.Objective: The aim of the study was to assess the effect of etidronate (bisphosphonate) on bone markers, inflammatory markers and RANKL/OPG in male subjects with active ankylosing spondylitis (AS). To explore the influence of etidronate on number and metabolic activity of proinflammatory subpopulation of blood monocytes CD14+CD16+ in AS patients. Material and methods: Etidronate (two 14-days cycles) was administered in a group of 25 men fulfiling New York AS criteria and having elevated ESR and/or CRP values. Osteocalcin (OC, formation marker), telopeptid β-CTx (CTX, resorption marker), soluble receptor activator of nuclear factor κB ligand (sRANKL) and osteoprotegerin (OPG) were assayed before and after each cycle of treatment. Results: The statistically significant decrease of CTX, OC and CRP, a non statistically significant elevation of OPG and reduction of sRANKL after etidronate treatment were observed. There were no correlation between bone markers and CRP, sRANKL and OPG as well as between CRP and sRANKL and OPG. Statistically significant correlation between sRANKL/OPG ratio before treatment and after 1st and 2nd cycle were found. No difference between number of proinflammatory monocytes (CD14+ CD16+) and classic monocytes (CD14++CD16–) were found before and after etidronate treatment with regard to the true number of cells in blood. Following PHA stimulation the avarage 80-fold increase in IL-10 production and only 40-fold rise in TNF-α were observed after 1. cycle of etidronate in PHA-stimulated peripheral blood mononuclear cells of patients, in comparison to cytokine production straight before the first run of etidronate. Conclusions: 1. Etidronate has anti-resorptive and anti-inflammatory properties in men with AS. 2. There was no correlation between CRP and bone markers, sRANKL, OPG and sRANKL/OPG ratio which may suggest a lack of relationship between inflammation and bone remodeling in AS. 3. Etidronate treatment does not influence the number of proinflammatory monocytes CD14+CD16+ in peripheral blood and increases the PHA-stimulated cytokine production by peripheral blood mononuclear cells, with prevalent immunosuppressive IL-10 synthesis