Cytokine Levels After Consumption of a Medicinal Agaricus blazei Murill‐Based Mushroom Extract,AndoSan™, in Patients with Crohn's Disease and Ulcerative Colitis in a Randomized Single‐Blinded Placebo‐Controlled Study

Ingestion of the Agaricus blazei Murill‐based mushroom extract AndoSan^? has been shown in randomized placebo‐controlled studies to improve symptoms in Crohn's disease (CD) and ulcerative colitis (UC) and also fatigue and quality of life in the latter patients. The aim was to examine whether this clinical impact of AndoSan^? intake could be explained by influence on foremost pro‐inflammatory cytokines in the patients. Fifty patients with symptomatic UC and CD were randomized and blinded for oral daily intake of AndoSan^? or placebo. Blood samples taken before (visit 1) and after 21 days’ (visit 3) consumption were analysed for cytokines IL‐1ß, IL‐2, IL‐4‐8, IL‐10, IL‐12‐13, IL‐17, G‐CSF, GM‐CSF, IFN‐γ, MCP‐1, MIP‐1ß and TNF‐α. Baseline cytokine levels were similar in CD and UC. In CD, cytokine levels at visit 1 versus visit 3 were unaltered within the AndoSan^? and the placebo groups. Only IL‐2 was significantly reduced at visit 3 in the Andosan^? compared with the placebo group. However, when combining IL‐1ß, IL‐6 and G‐CSF in the patients with CD, the cytokine levels were significantly lower in the AndoSan^TM ‐ versus the placebo group, visit 3. In UC, levels of IL‐2, IL‐5 and MIP‐1ß were reduced within the AndoSan^? group. IL‐5 was also reduced at visit 3 compared with placebo. Generally, the effect on reduction in systemic cytokine levels by consumption of AndoSan^? was limited and supported only marginally anti‐inflammatory effects in these patients. Therefore, other explanations behind the clinical anti‐inflammatory effects than the contribution of cytokines seem more pertinent, including anti‐allergic and antioxidant activities.     

Serum concentrations of cytokines in infants with retinopathy of prematurity

Retinopathy of prematurity (ROP) is the leading cause of blindness in preterm infants. In this study, we investigated the cytokine levels in cord blood of normal preterm neonates and preterm infants developed ROP. Serum levels of 10 cytokines in umbilical cord blood were measured by multiplex protein arrays from 62 healthy preterm neonates and 30 preterm neonate cases who developed ROP at later stage. Results showed that serum levels of cytokines including interleukin 7 (IL‐7), monocyte chemotactic protein‐1 (MCP‐1), macrophage inflammatory protein 1 alpha (MIP‐1α), and macrophage inflammatory protein 1 beta (MIP‐1β) were significantly increased in cases who developed ROP than in healthy preterm neonates (3.5‐fold, 3.2‐fold, 3.4‐fold, and 2.1‐fold, respectively), whereas levels of these four cytokines did not reveal any significant differences between healthy preterm infants and normal infants. When comparing the expression of cytokines in ROP patients with different clinical parameters, ROP cases whose gestational age at delivery earlier than 29.0 weeks demonstrated increased levels of MCP‐1 and MIP‐1β than those later than 29.0 weeks (p < 0.05). Also, ROP cases with birth weight less than 1.28 kg revealed significantly higher level of MIP‐1β than those who were heavier than 1.28 kg (p < 0.05). These data indicated that levels of IL‐7, MCP‐1, MIP‐1α, and MIP‐1β were associated with increased risk of ROP, in which MIP‐1β may be further correlated with the severity of ROP.     

Distinct expression of interleukin (IL)‐36α, β and γ, their antagonist IL‐36Ra and IL‐38 in psoriasis,rheumatoid arthritis and Crohn's disease

Interleukin (IL)‐36α, IL‐36β and IL‐36γ are expressed highly in skin and are involved in the pathogenesis of psoriasis, while the antagonists IL‐36Ra or IL‐38, another potential IL‐36 inhibitor, limit uncontrolled inflammation. The expression and role of IL‐36 cytokines in rheumatoid arthritis (RA) and Crohn's disease (CD) is currently debated. Here, we observed that during imiquimod‐induced mouse skin inflammation and in human psoriasis, expression of IL‐36α, γ and IL‐36Ra, but not IL‐36β and IL‐38 mRNA, was induced and correlated with IL‐1β and T helper type 17 (Th17) cytokines (IL‐17A, IL‐22, IL‐23, CCL20). In mice with collagen‐induced arthritis and in the synovium of patients with RA, IL‐36α, β, γ, IL‐36Ra and IL‐38 were all elevated and correlated with IL‐1β, CCL3, CCL4 and macrophage colony‐stimulating factor (M‐CSF), but not with Th17 cytokines. In the colon of mice with dextran sulphate sodium‐induced colitis and in patients with CD, only IL‐36α, γ and IL‐38 were induced at relatively low levels and correlated with IL‐1β and IL‐17A. We suggest that only a minor subgroup of patients with RA (17–29%) or CD (25%) had an elevated IL‐36 agonists/antagonists ratio, versus 93% of patients with psoriasis. By immunohistochemistry, IL‐36 cytokines were produced by various cell types in skin, synovium and colonic mucosa such as keratinocytes, CD68⁺ macrophages, dendritic/Langerhans cells and CD79α⁺ plasma cells. In primary cultures of monocytes or inflammatory macrophages (M1), IL‐36β and IL‐36Ra were produced constitutively, but IL‐36α, γ and IL‐38 were produced after lipopolysaccharide stimulation. These distinct expression profiles may help to explain why only subgroups of RA and CD patients have a potentially elevated IL‐36 agonists/antagonists ratio.     

Effect of an Extract Based on the Medicinal Mushroom Agaricus blazei Murill on Release of Cytokines, Chemokines and Leukocyte Growth Factors in Human Blood Ex Vivo and In Vivo

An immunostimulatory extract based on the medicinal mushroom Agaricus blazei Murill (AbM) has been shown to stimulate mononuclear phagocytes in vitro to produce pro-inflammatory cytokines, and to protect against lethal peritonitis in mice. The present aim was to study the effect of AbM on release of several cytokines in human whole blood both after stimulation ex vivo and in vivo after oral intake over several days in healthy volunteers. The 17 signal substances examined were; T helper 1 (Th1) cytokines [interleukin (IL)-2, interferon (IFN)-γ and IL-12], T helper 2 cytokines (IL-4, IL-5 and IL-13), pleiotropic (IL-7, IL-17), pro-inflammatory [IL-1β, IL-6, tumour necrosis factor (TNF)-α (mainly produced by Th1 cells)] – and anti-inflammatory (IL-10) cytokines, chemokines [IL-8, macrophage inhibitory protein (MIP)-1β and monocyte chemoattractant protein (MCP)-1] and leukocyte growth factors [granulocyte colony-stimulating factor (G-CSF), granulocyte/macrophage colony stimulating factor]. After stimulation of whole blood ex vivo with 0.5–5.0% of a mushroom extract, AndoSan™ mainly containing AbM , there was a dose-dependent increase in all the cytokines studied, ranging from two to 399-fold (TNF-α). However, in vivo in the eight volunteers who completed the daily intake (60 ml) of this AbM extract for 12 days, a significant reduction was observed in levels of IL-1β (97%), TNF-α (84%), IL-17 (50%) and IL-2 (46%). Although not significant, there was a trend towards reduced levels for IL-8, IFN-γ and G-CSF, whilst those of the remaining nine cytokines tested, were unaltered. The discrepant results on cytokine release ex vivo and in vivo may partly be explained by the antioxidant activity of AbM in vivo and limited absorption of its large, complex and bioactive β-glucans across the intestinal mucosa to the reticuloendothelial system and blood.     

CXC and CC chemokines induced in human renal epithelial cells by inflammatory cytokines

Human renal epithelial cells might play an important role during the allograft rejection by producing chemokines in response to proinflammatory cytokines such as tumor necrosis factor (TNF)‐α and interleukin (IL)‐1β produced by endothelial and epithelial cells early after transplantation. The production of chemokines allows inflammatory cells to be drawn into the kidney graft and therefore plays a critical role in the pathophysiologic processes that lead to the rejection of renal transplant. In this process, two chemokine superfamilies, the CC and the CXC chemokines, are the most important. The CC chemokines target mainly monocytes and T lymphocytes, while most of the CXC chemokines attract neutrophils. We showed in our study that in vitro, in unstimulated cells, basal mRNA expression of CXC chemokines (Groα, Groβ, Groγ, ENA‐78 and GCP‐2, IL‐8) that attract neutrophils was detectable and expression of these genes and chemokine release were increased in TNF‐α‐ and IL‐1β‐induced renal epithelial cells. Most of the CC chemokines [monocyte chemotactic protein‐1 (MCP‐1), macrophage Inflammatory protein 1 beta (MIP‐1β), regulated upon activation, normal T cell expressed and secreted (RANTES) and macrophage inflammatory protein (MIP‐3α)] showed detectable mRNA expression only after stimulation with proinflammatory cytokines and not in control cells. TNF‐α seems to induce preferably the expression of RANTES, MCP‐1, interferon‐inducible protein (IP‐10) and Interferon‐Inducible T‐cell Alpha Chemoattractant (I‐TAC), while IL‐1β induces mainly IL‐8 and epithelial neutrophil‐activating peptide 78 (ENA‐78).     

FluoroSpot Analysis of TLR‐Activated Monocytes Reveals Several Distinct Cytokine‐Secreting Subpopulations

Monocytes have long been considered a heterogeneous group of cells both in terms of morphology and function. In humans, three distinct subsets have been described based on their differential expression of the cell surface markers CD14 and CD16. However, the relationship between these subsets and the production of cytokines has for the most part been based on ELISA measurements, making it difficult to draw conclusions as to their functional profile on the cellular level. In this study, we have investigated lipoteichoic acid (LTA)‐ and lipopolysaccharide (LPS)‐induced cytokine secretion by monocytes using the FluoroSpot technique. This method measures the number of cytokine‐secreting cells on the single‐cell level and uses fluorescent detection, allowing for the simultaneous analysis of two cytokines from the same population of isolated cells. By this approach, human monocytes from healthy volunteers could be divided into several subgroups as IL‐1β, IL‐6, TNF‐α and MIP‐1β were secreted by larger populations of responding cells (25.9–39.2%) compared with the smaller populations of GM‐CSF (9.1%), IL‐10 (1.3%) and IL‐12p40 (1.2%). Furthermore, when studying co‐secretion in FluoroSpot, an intricate relationship between the monocytes secreting IL‐1β and/or IL‐6 and those secreting TNF‐α, MIP‐1β, GM‐CSF, IL‐10 and IL‐12p40 was revealed. In this way, dissecting the secretion pattern of the monocytes in response to TLR‐2 or TLR‐4 stimulation, several subpopulations with distinct cytokine‐secreting profiles could be identified.     

TL1A as a Potential Local Inducer of IL17A Expression in Colon Mucosa of Inflammatory Bowel Disease Patients

Interaction between TL1A and death receptor 3 (DR3) co‐stimulates T cells, induces production of several pro‐inflammatory cytokines and has been linked to pathogenesis of inflammatory bowel disease (IBD). This study aimed to establish a link between expression of TL1A and selected TL1A‐induced pro‐inflammatory cytokines involved in IBD pathogenesis (IL‐4, IL‐13, IL‐17A and IFN‐γ) and to investigate a connection between serum concentration of TL1A in patients with IBD and activation of peripheral blood T cells. Elevated levels of IL‐4 (2.91‐fold) and IL‐13 (4.05‐fold) mRNA were detected in the inflamed colon mucosa of patients with ulcerative colitis (UC), IFN‐γ mRNA was upregulated (3.23‐fold) in the inflamed colon mucosa of patients with Crohn's disease (CD), whereas upregulation of IL‐17A and TL1A mRNA was present in the inflamed colon mucosa of patients with both CD and UC (IL‐17A: 4.48‐fold and 2.74‐fold, TL1A: 3.19‐fold and 3.22‐fold, respectively) vs. control subjects. We did not detect any changes in DR3 mRNA expression in the investigated groups of patients. TL1A mRNA level in colon mucosa of patients with IBD correlated only with the level of IL‐17A mRNA but no other investigated cytokines. In colon mucosa, expression of TL1A and DR3 was localized to enterocytes and lamina propria mononuclear cells. We did not find any correlation between serum concentrations of TL1A and IL‐17A or changes of CD4(+) or CD8(+) lymphocytes phenotype in patients with IBD. Therefore, our data indicate that TL1A may contribute to pathogenesis of IBD via local but not systemic induction of IL‐17A but not IL‐4, IL‐13 or IFN‐γ.     

Absence of a role for interleukin‐13 in inflammatory bowel disease

IL‐13 has been implicated in the pathogenesis of ulcerative colitis (UC), and may have a role in animal models of gut fibrosis. We studied the involvement of IL‐13 in inflammation and fibrosis in UC and Crohn's disease (CD). Intestinal biopsies and anti‐CD3/CD28‐ or anti‐CD2/CD28‐stimulated lamina propria mononuclear cells from UC and CD patients and control subjects were cultured, and IL‐13, IL‐4, IL‐5, IL‐17A and IFN‐γ production was measured. Mucosal IL‐13‐producing cells were characterised by flow cytometry. Gut explants from strictured CD, non‐strictured CD and healthy donors were cultured ex vivo, and secreted IL‐13, IL‐1β and collagen were measured. IL‐13 production by mucosal explants and activated lamina propria mononuclear cells did not differ between CD, UC and control subjects, and was at least a log lower than IFN‐γ and IL‐17A. IL‐13‐producing cells, and in particular natural killer T cells, were uniformly low in all groups. IL‐4 and IL‐5 were undetectable in culture supernatants. Explants of CD strictures produced low amounts of IL‐13, whereas IL‐1β and collagen were elevated. We could not confirm that UC or strictured CD are associated with elevated IL‐13 production. These data suggest that an anti‐IL‐13 Ab would not be an appropriate therapeutic strategy in inflammatory bowel disease.     

Occupational exposure to diesel engine exhaust and serum cytokine levels

The International Agency for Research on Cancer has classified diesel engine exhaust (DEE) as a human lung carcinogen. Given that inflammation is suspected to be an important underlying mechanism of lung carcinogenesis, we evaluated the relationship between DEE exposure and the inflammatory response using data from a cross‐sectional molecular epidemiology study of 41 diesel engine testing workers and 46 unexposed controls. Repeated personal exposure measurements of PM_2.5 and other DEE constituents were taken for the diesel engine testing workers before blood collection. Serum levels of six inflammatory biomarkers including interleukin (IL)‐1, IL‐6, IL‐8, tumor necrosis factor (TNF)‐α, macrophage inflammatory protein (MIP)‐1β, and monocyte chemotactic protein (MCP)‐1 were analyzed in all subjects. Compared to unexposed controls, concentrations of MIP‐1β were significantly reduced by ～37% in DEE exposed workers (P < 0.001) and showed a strong decreasing trend with increasing PM_2.5 concentrations in all subjects (P_trend < 0.001) as well as in exposed subjects only (P_trend = 0.001). Levels of IL‐8 and MIP‐1β were significantly lower in workers in the highest exposure tertile of PM_2.5 (>397 µg/m³) compared to unexposed controls. Further, significant inverse exposure‐response relationships for IL‐8 and MCP‐1 were also found in relation to increasing PM_2.5 levels among the DEE exposed workers. Given that IL‐8, MIP‐1β, and MCP‐1 are chemokines that play important roles in recruitment of immunocompetent cells for immune defense and tumor cell clearance, the observed lower levels of these markers with increasing PM_2.5 exposure may provide insight into the mechanism by which DEE promotes lung cancer. Environ. Mol. Mutagen. 59:144–150, 2018. © 2017 Wiley Periodicals, Inc.     

Comparison of immunological adjuvants

In this study, several innate immunological adjuvants and related compounds were compared with respect to complement activation in serum and induction of cytokine release in whole blood samples using immunoassays. As found, simple lipids had no effect on the complement system or on cytokine release, whereas lipopolysaccharides induced prominent release of pro‐inflammatory cytokines (IL1β, TNF and IFNγ) without affecting the complement system, except for one, which activated the lectin pathway (LP). Moreover, saponin induced IL1β and MCP1 release and did not affect the complement system. The polysaccharide inulin exhausted the alternative pathway (AP) completely without affecting the LP and the classical pathway (CP), whereas zymosan exhausted the AP and had a major effect on the LP and CP as well. Peptidoglycans mainly affected the LP. Inulin, agarose and cellulose induced IL1β and MCP1 release, while dextran had no effect on cytokine secretion. Zymosan mainly induced IL1β release. The inorganic compound aluminium hydroxide, Al(OH)₃, activated the complement system very efficiently (all three pathways) but only induced MCP1 release. Other compounds tested had minor/individual effects. Collectively, well‐known adjuvants, such as aluminum hydroxide, activated the complement system and/or induced pro‐inflammatory cytokine release. Since complement activation generates anaphylactic peptides, a simple definition of an (innate) immunological adjuvant can be inferred: it activates the (innate) immune system by complement activation and/or release of cytokines so as to attract cells of the adaptive immune system.     

Induction, function and regulation of IL-17-producing T cells

Recent reports have provided convincing evidence that IL‐17‐producing T cells play a key role in the pathogenesis of organ‐specific autoimmune diseases, a function previously attributed exclusively to IFN‐γ‐secreting Th1 cells. Furthermore, it appears that IL‐17‐producing T cells can also function with Th1 cells to mediate protective immunity to pathogens. Although much of the focus has been on IL‐17‐secreting CD4⁺ T cells, termed Th17 cells, CD8⁺ T cells, γδ T cells and NKT cells are also capable of secreting IL‐17. The differentiation of Th17 cells from naïve T cells appears to involve signals from TGF‐β, IL‐6, IL‐21, IL‐1β and IL‐23. Furthermore, IL‐1α or IL‐1β in synergy with IL‐23 can promote IL‐17 secretion from memory T cells. The induction or function of Th17 cells is regulated by cytokines secreted by the other major subtypes of T cells, including IFN‐γ, IL‐4, IL‐10 and at high concentrations, TGF‐β. The main function of IL‐17‐secreting T cells is to mediate inflammation, by stimulating production of inflammatory cytokines, such as TNF‐α, IL‐1β and IL‐6, and inflammatory chemokines that promote the recruitment of neutrophils and macrophages.     

Increased cytokines,chemokines and soluble adhesion molecules in exhaled breath condensate of asthmatic children

Background Airway inflammation in asthma is characterized by the production of cytokines, chemokines and soluble adhesion molecules. The assessment of these inflammatory biomarkers in exhaled breath condensate (EBC) is hampered by low detection rates. However, the use of a glass condenser system combined with a sensitive analytical technique may increase the possibility to assess these biomarkers in EBC in a reliable way. Objective (1) To assess the detection rates of cytokines (IL‐1α, ‐1β, ‐2, ‐4, ‐5, ‐6, ‐10, ‐12p70, ‐13, ‐18, IFN‐γ, TNF‐α), chemokines [MIP1α (CCL3), MIF, eotaxin (CCL11), RANTES (CCL5), IP10 (CXCL10), IL8 (CXCL8), MCP1] and soluble adhesion molecules [soluble intercellular adhesion molecule (sICAM), soluble vascular adhesion molecule (sVCAM)] in EBC of children with asthma and healthy control children;
(2) To study the differences in the biomarker concentration between children with asthma and controls. Methods Sixty children were included: 31 asthmatics (71% atopic) and 29 controls. Exhaled breath condensate was collected using a glass condenser system. The inflammatory markers (IM) were analysed using multiplex immunoassay technology. Results Detection percentages of cytokines, chemokines and adhesion molecules ranged from 94% to 100%, except for eotaxin (CCL11) and RANTES (CCL5) (detection rates of 10% and 45% in healthy controls, respectively). The intra‐subject variability of biomarkers in EBC in the group as a whole ranged from 5.2% to 35.0%. In asthmatics, the levels of cytokines (IL‐2, ‐4, ‐5, ‐6, ‐13, IFN‐γ), chemokines (MIP1α [CCL3], MIF, RANTES [CCL5], IP10 [CXCL10], IL8 [CXCL8], MCP1) and adhesion molecules (sICAM, sVCAM) were significantly increased in comparison with controls (P<0.05). Conclusion If collected with a glass condenser and analysed by multiplex immunoassay technology, cytokines, chemokines and soluble adhesion molecules can be reliably demonstrated in EBC of children. Most of these IM were elevated in EBC of asthmatics compared with controls. Cite this as: C. M. H. H. T. Robroeks, G. T. Rijkers, Q. Jöbsis, H. J. E. Hendriks, J. G. M. C. Damoiseaux, L. J. I. Zimmermann, O. P. van Schayck, E. Dompeling, Clinical & Experimental Allergy, 2010 (40) 77–84.     

Chemotactic activity on mononuclear cells in the cerebrospinal fluid of patients with viral meningitis is mediated by interferon-γ inducible protein-10 and monocyte chemotactic protein-1

In viral meningitis the inflammatory response involves activated T cells and monocytes which are recruited into the subarachnoid space. To identify the chemotactic signals attracting the cells to the site of infection in the meninges, we measured the levels of two CXC chemokines, interferon-γ (IFN-γ) inducible protein (IP)-10 and monokine induced by IFN-γ, four CC chemokines, monocyte chemotactic protein (MCP)-1, RANTES, macrophage inflammatory protein (MIP)-1α and MIP-1β, as well as the cytokines interleukin (IL)-15 and IL-16 in the cerebrospinal fluid (CSF) of patients suffering from viral meningitis. The results point to an involvement of two chemokines, MCP-1 and IP-10, since (1) unlike the other cytokines, MCP-1 and IP-10 were present in 97 % and 79 % of the CSF, respectively, at concentrations sufficient to induce chemotaxis of mononuclear cells; (2) more than 90 % of the CSF of viral meningitis induced chemotaxis of peripheral blood mononuclear cells (PBMC) and all of them induced chemotaxis of activated T cells, and (3) the CSF-mediated chemotaxis of PBMC was inhibited by anti-MCP-1 antibodies and chemotaxis of activated T cells was abolished by the combination of anti-MCP-1 and anti-IP-10 antibodies. Our data provide evidence that MCP-1 and IP-10 lead to accumulation of activated T cells and monocytes in the CSF compartment in viral meningitis.     

Can medicinal mushrooms have prophylactic or therapeutic effect against COVID‐19 and its pneumonic superinfection and complicating inflammation?

Medicinal mushrooms have documented effects against different diseases, including infections and inflammatory disorders. The related Basidiomycota Agaricus blazei Murill (AbM), Hericium erinaceus (HE), and Grifola frondosa (GF) have been shown to exert antimicrobial activity against viral agents, Gram‐positive and Gram‐negative bacteria, and parasites in vitro and in vivo. Since the mechanism is immunomodulatory and not antibiotical, the mushrooms should be active against multi‐drug resistant microbes as well. Moreover, since these Basidiomycota also have anti‐inflammatory properties, they may be suited for treatment of the severe lung inflammation that often follows COVID‐19 infection. An AbM‐based mushroom extract (Andosan?), also containing HE and GF, has been shown to significantly reduce bacteraemia and increase survival in mice with pneumococcal sepsis, and to improve symptoms and quality of life in IBD patients via an anti‐inflammatory effect. Hence, such mushroom extracts could have prophylactic or therapeutic effect against the pneumonic superinfection and severe lung inflammation that often complicates COVID‐19 infection. Here, we review antimicrobial and anti‐inflammatory properties of AbM, HE and GF mushrooms, which could be used for the battle against COVID‐19.     

An Extract of the Medicinal Mushroom Agaricus blazei Murill Differentially Stimulates Production of Pro-inflammatory Cytokines in Human Monocytes and Human Vein Endothelial Cells in vitro

An extract of the edible mushroom Agaricus blazei Murill (AbM) has known antitumor and anti-infection properties, probably mainly by stimulating mononuclear phagocytes of the native immune system. The aim of this work was to study the effect of AbM on the production by human monocytes and human umbilical vein endothelial cells (EC) of pro-inflammatory cytokines (IL-1β, IL-6, IL-8, TNFα), the anti-inflammatory/T regulatory cytokine IL-10 and the pro-Th1 cytokine IL-12. AbM, in concentrations from 1–15%, induced a considerable and dose-dependent increase in production of IL-8, IL-6, TNFα and IL-1β in monocyte cultures. The biosynthesis reached a plateau at a concentration of 10% of AbM, and was most pronounced for the three former cytokines. AbM did also dose-dependently stimulate EC production of IL-8,IL-6 and TNFα, but at lower levels compared with the monocytes. AbM did neither induce synthesis of cytokines IL-10 nor IL-12 in monocytes or EC. Our results demonstrate the differential effect of AbM stimulation on the magnitude of pro-inflammatory cytokines produced by monocytes and EC.     

Fatty acids modulate cytokine and chemokine secretion of stimulated human whole blood cultures in diabetes

Fatty acids, uric acid and glucose are thought to contribute to subclinical inflammation associated with diabetes mellitus. We tested whether co‐incubation of free fatty acids and uric acid or glucose influences the secretion of immune mediators from stimulated human whole blood in vitro. Fresh whole blood samples from 20 healthy subjects, 20 patients with type 1 diabetes and 23 patients with type 2 diabetes were incubated for 24 h with palmitic acid (PAL), linolenic acid (LIN) or eicosapentaenoic acid (EPA) alone or together with elevated concentrations of uric acid or glucose. Concentrations of proinflammatory cytokines interleukin (IL)‐1β, IL‐2, IL‐12(p70), IL‐18, IFN‐γ, of regulatory cytokines IL‐4, IL‐10, IL‐17 and chemokine CCL2 (MCP‐1) were measured by multiplex‐bead technology from supernatants. Co‐incubation of fatty acids with uric acid resulted in a significant reduction of IL‐10, IL‐12(p70), IFN‐γ and CCL2 (MCP‐1) concentrations in supernatants compared to incubation with uric acid alone (P < 0·0001). In contrast, IL‐18 was up‐regulated upon co‐stimulation with fatty acids and uric acid. Similarly, co‐incubation of fatty acids with glucose diminished secretion of IL‐10, IFN‐γ and CCL2 (monocyte chemotactic protein‐1), while IL‐8 was up‐regulated (P < 0·001). Samples from healthy and diabetic subjects did not differ after adjustment for age, sex, body mass index and diabetes type. All three fatty acids similarly influenced whole blood cytokine release in vitro and modulated uric acid or glucose‐stimulated cytokine secretion. Although the ω‐3‐fatty acid EPA showed slightly stronger effects, further studies are required to elaborate the differential effects of PAL, LIN and EPA on disease risk observed previously in epidemiological studies.     

Differential Expression of CD30 on CD3 T Lymphocytes in Patients with Systemic Lupus Erythematosus

Systemic lupus erythematosus (SLE) is an autoimmune systemic disease caused as a result of an imbalance of Th1‐/Th2‐type cytokines. The soluble form of CD30 (CD30s) released from peripheral blood cells has been described as a marker of active disease in Th2‐type immune response as in SLE. However, the expression of CD30 on CD3 T lymphocytes from patients with SLE has not been studied yet. Therefore, we have addressed our study to attempt this issue, studying CD30 expression by flow cytometry on CD3 T lymphocytes and CD4/CD8 subsets in samples from SLE patients mainly with lupus nephritis. In parallel, we have determined the production of the cytokines IL‐4 (Th2), IFNγ (Th1), IL‐10 and TGFβ by intracellular staining. Differences between positive CD30 T cells in healthy controls and patients with SLE were found, with a higher percentage of CD30‐expressing T cells in patients with SLE (P = 0.001). In contrast to healthy controls, CD30 was mainly expressed on CD8 T cells from patients with SLE. The intracellular cytokine staining showed that TGFβ is the main cytokine expressed in CD3 T cells from patients with SLE. In addition to this, we have found a positive correlation between CD30‐expressing T cells and IL‐4, IFNγ, and immunosuppressive cytokines (IL‐10 and TGFβ) (P < 0.05). These results suggest that CD30 could play a role in the pathogenesis of SLE and its expression on CD3 T lymphocytes is not restricted only to Th2‐type response.     

Bromelain treatment decreases secretion of pro-inflammatory cytokines and chemokines by colon biopsies in vitro

Oral bromelain has been anecdotally reported to decrease inflammation in ulcerative colitis (UC). Proteolytically active bromelain is known to decrease expression of mRNAs encoding pro-inflammatory cytokines by human leukocytes in vitro. To assess the effect of bromelain on mucosal secretion of cytokines in inflammatory bowel disease (IBD), endoscopic colon biopsies from patients with UC, Crohn's disease (CD), and non-IBD controls were treated in vitro with bromelain or media, then cultured. Secretion of pro-inflammatory cytokines and chemokines was measured. Significant increases in granulocyte colony-stimulating factor (G-CSF), interferon (IFN)-gamma, interleukin (IL)-1beta, IL-6, and tumor necrosis factor (TNF) were detected in the media from actively inflamed areas in UC and CD as compared with non-inflamed IBD tissue and non-IBD controls. In vitro bromelain treatment decreased secretion of G-CSF, granulocyte-macrophage colony-stimulating factor (GM-CSF), IFN-gamma, CCL4/macrophage inhibitory protein (MIP)-1beta, and TNF by inflamed tissue in IBD. Bromelain may be a novel therapy for IBD.     

A possible link between recurrent upper respiratory tract infections and lower cytokine production in patients with Q fever fatigue syndrome

Besides fatigue, many Q fever fatigue syndrome (QFS) patients also complain of frequently recurring upper respiratory tract infections with severe symptoms. We investigated whether immunologic dysregulation contributes to these complaints. Cytokine and chemokine production was measured after stimulating monocytes of QFS patients and age‐ and sex‐matched healthy controls with LPS and several viral ligands. The H3K4me3 mark of open chromatin was measured at the promoter regions of cytokines and chemokines that differed significantly from healthy controls. Monocytes of QFS patients produced significantly less TNF‐α (p = 0.032), IL‐1β (0.004, 0.024, and 0.008), IL‐6 (0.043), RANTES (0.033), IP‐10 (0.049), MCP‐1 (0.022), IL‐ 13 (0.029), and IL‐10 (0.026) than healthy controls when stimulated with various ligands. H3K4me3 expression was significantly lower in QFS patients than in healthy controls on the promoter regions of IL‐1β (p = 0.004), MCP‐1 (<0.001 and <0.001), IP‐10 (<0.001), IL‐10 (0.041), and IL‐13 (<0.001, <0.001, and 0.001). QFS patients showed diminished cytokine responses to various stimuli compared to age‐ and sex‐matched healthy controls, likely due to epigenetic remodeling and long‐term memory as a result from the acute Q fever infection. This might explain the upper respiratory tract ailments in QFS.