Soluble CD14 Concentrations Research Articles

Hemorrhagic transformation in patients with large-artery atherosclerotic stroke is associated with the gut microbiota and lipopolysaccharide.

Abstract JOURNAL/nrgr/04.03/01300535-202407000-00032/figure1/v/2023-11-20T171125Z/r/image-tiff Hemorrhagic transformation is a major complication of large-artery atherosclerotic stroke (a major ischemic stroke subtype) that worsens outcomes and increases mortality. Disruption of the gut microbiota is an important feature of stroke, and some specific bacteria and bacterial metabolites may contribute to hemorrhagic transformation pathogenesis. We aimed to investigate the relationship between the gut microbiota and hemorrhagic transformation in large-artery atherosclerotic stroke. An observational retrospective study was conducted. From May 2020 to September 2021, blood and fecal samples were obtained upon admission from 32 patients with first-ever acute ischemic stroke and not undergoing intravenous thrombolysis or endovascular thrombectomy, as well as 16 healthy controls. Patients with stroke who developed hemorrhagic transformation (n = 15) were compared to those who did not develop hemorrhagic transformation (n = 17) and with healthy controls. The gut microbiota was assessed through 16S ribosomal ribonucleic acid sequencing. We also examined key components of the lipopolysaccharide pathway: lipopolysaccharide, lipopolysaccharide-binding protein, and soluble CD14. We observed that bacterial diversity was decreased in both the hemorrhagic transformation and non-hemorrhagic transformation group compared with the healthy controls. The patients with ischemic stroke who developed hemorrhagic transformation exhibited altered gut microbiota composition, in particular an increase in the relative abundance and diversity of members belonging to the Enterobacteriaceae family. Plasma lipopolysaccharide and lipopolysaccharide-binding protein levels were higher in the hemorrhagic transformation group compared with the non-hemorrhagic transformation group. lipopolysaccharide, lipopolysaccharide-binding protein, and soluble CD14 concentrations were associated with increased abundance of Enterobacteriaceae. Next, the role of the gut microbiota in hemorrhagic transformation was evaluated using an experimental stroke rat model. In this model, transplantation of the gut microbiota from hemorrhagic transformation rats into the recipient rats triggered higher plasma levels of lipopolysaccharide, lipopolysaccharide-binding protein, and soluble CD14. Taken together, our findings demonstrate a noticeable change in the gut microbiota and lipopolysaccharide-related inflammatory response in stroke patients with hemorrhagic transformation. This suggests that maintaining a balanced gut microbiota may be an important factor in preventing hemorrhagic transformation after stroke.

New Insights on End-Stage Renal Disease and Healthy Individual Gut Bacterial Translocation: Different Carbon Composition of Lipopolysaccharides and Different Impact on Monocyte Inflammatory Response.

Chronic kidney disease induces disruption of the intestinal epithelial barrier, leading to gut bacterial translocation. Here, we appreciated bacterial translocation by analyzing circulating lipopolysaccharides (LPS) using two methods, one measuring only active free LPS, and the other quantifying total LPS as well as LPS lipid A carbon chain length. This was done in end-stage renal disease (ESRD) patients and healthy volunteers (HV). We observed both higher LPS concentration in healthy volunteers and significant differences in composition of translocated LPS based on lipid A carbon chain length. Lower LPS activity to mass ratio and higher concentration of high-density lipoproteins were found in HV, suggesting a better plasma capacity to neutralize LPS activity. Higher serum concentrations of soluble CD14 and pro-inflammatory cytokines in ESRD patients confirmed this hypothesis. To further explore whether chronic inflammation in ESRD patients could be more related to LPS composition rather than its quantity, we tested the effect of HV and patient sera on cytokine secretion in monocyte cultures. Sera with predominance of 14-carbon chain lipid A-LPS induced higher secretion of pro-inflammatory cytokines than those with predominance of 18-carbon chain lipid A-LPS. TLR4 or LPS antagonists decreased LPS-induced cytokine production by monocytes, demonstrating an LPS-specific effect. Thereby, septic inflammation observed in ESRD patients may be not related to higher bacterial translocation, but to reduced LPS neutralization capacity and differences in translocated LPS subtypes.

Ambient air pollution and cardiovascular disease in Ugandan adolescents with perinatally acquired HIV: a cross-sectional study

BackgroundAir pollution is known to induce systemic inflammation and contribute to cardiovascular disease. The effects of ambient air pollution on adolescents living with perinatally acquired HIV (PHIV) in Africa has been understudied. In this study, we aim to investigate the relationship between particulate matter, a common proxy indicator for air pollution, and markers of inflammation, monocyte activation, and cardiovascular disease risk. MethodsWe recruited a cohort of PHIV-positive and HIV-negative adolescents who were from around Kampala, aged 10–18 years, had no known active infections, and who lived within 40 km of an PM2·5 monitoring site. Adolescents with PHIV were on ART with HIV-1 RNA concentrations ≤400 copies/mL. Daily ambient concentrations of particulate matter (PM2·5), a proxy for air pollution, were measured with continuous central site monitoring using a Beta Attenuation Monitor or E-Samplers from the GeoHealth Hub. We measured carotid intima media thickness (IMT) using ultrasound, plasma concentrations of soluble CD14 and CD163 as markers of monocyte activation, plasma markers of systemic inflammation (hsCRP, IL6, sTNFRI), oxidised lipids, and intestinal permeability (zonulin). FindingsOne hundred and nineteen participants were included (69 with PHIV, 50 were HIV-negative). Median age was 12·7 years (IQR 11·4–14·2) years, 55% were female and 63% lived below the extreme poverty line (living with <US$1·90 a day). Median CD4+ cell count was 962 cells/μL (643–1313), 82% had viral load <20 copies/mL and median ART duration was 12·1 years (9·7–13·3) years. Overall median daily PM2·5 exposure was 29·08 μg/m3 (23·40–41·70), considered a moderate health risk (WHO definition). There was no significant difference in exposure of PM2·5 between groups (p=0·073), nor between sexes (p=0·099). PM2·5 concentrations significantly correlated with intestinal permeability (zonulin; r=0·43, p<0·001), monocyte activation (soluble CD163: r=0·25, p=0·053), and IMT (r=0·35, p=0·004) in participants with PHIV but not in those who were HIV-negative (p≥0·05 for all three correlations in HIV-negative children). In multivariable quantile regression models, after adjusting for age, sex, and, separately, soluble CD163 and zonulin, daily PM2·5 concentrations remained associated with IMT (β=0·005, 95%CI [0·001–0·009], p=0·007 and β=0·004, 95%CI [0·001–0·008], p=0·014, respectively) in children with PHIV. InterpretationPM2·5 exposure was associated with elevated subclinical vascular disease in adolescents with PHIV but not HIV-negative adolescents, despite there being no significant difference in exposure between these two groups. Our findings suggest the combined effects of HIV infection and air pollution may amplify and contribute to early development of cardiovascular disease. FundingNICHD K23HD088295 (SDF); University Hospitals Cleveland Medical Center; The Clinical and Translational Science Collaborative of Cleveland; and UL1TR002548 from the National Center for Advancing Translational Sciences.

High-Dose Neonatal Vitamin A Supplementation to Bangladeshi Infants Increases the Percentage of CCR9-Positive Treg Cells in Infants with Lower Birthweight in Early Infancy, and Decreases Plasma sCD14 Concentration and the Prevalence of Vitamin A Deficiency at Two Years of Age

Vitamin A (VA) stores are low in early infancy and may impair development of the immune system. This study determined if neonatal VA supplementation (VAS) affects the following: 1) development of regulatory T (Treg) cells; 2) chemokine receptor 9 (CCR9) expression, which directs mucosal targeting of immune cells; and 3) systemic endotoxin exposure as indicated by changed plasma concentrations of soluble CD14 (sCD14). Secondarily, VA status, growth, and systemic inflammation were investigated. In total, 306 Bangladeshi infants were randomly assigned to receive 50,000 IU VA or placebo (PL) within 48h of birth, and immune function was assessed at 6 wk, 15 wk, and 2 y. Primary outcomes included the following: 1) peripheral blood Treg cells; 2) percentage of Treg, T, and B cells expressing CCR9; and 3) plasma sCD14. Secondary outcomes included the following: 4) VA status measured using the modified relative dose-response (MRDR) test and plasma retinol; 5) infant growth; and 6) plasma C-reactive protein (CRP). Statistical analysis identified group differences and interactions with sex and birthweight. VAS increased (P=0.004) the percentage of CCR9+ Treg cells (13.2± 1.37%) relative to PL (9.17± 1.15%) in children below the median birthweight but had the opposite effect (P=0.04) in those with higher birthweight (VA, 9.13± 0.89; PL, 12.1± 1.31%) at 6 and 15 wk (values are combined mean±SE). VAS decreased (P=0.003) plasma sCD14 (1.56± 0.025mg/L) relative to PL (1.67± 0.032mg/L) and decreased (P=0.034) the prevalence of VA deficiency (2.3%) relative to PL (9.2%) at 2 y. Neonatal VAS enhanced mucosal targeting of Treg cells in low-birthweight infants. The decreased systemic exposure to endotoxin and improved VA status at 2 y may have been due to VA-mediated improvements in gut development resulting in improved barrier function and nutrient absorption. This trial was registered at clinicaltrials.gov as NCT01583972 and NCT02027610.

Soluble CD14 Induces Pro-inflammatory Cytokines in Rheumatoid Arthritis Fibroblast-Like Synovial Cells via Toll-Like Receptor 4.

Objectives: Synovial fluids of rheumatoid arthritis (RA) patients commonly contain high concentrations of soluble CD14 (sCD14). To investigate its potential role in RA pathogenesis, we tested whether sCD14 binding transmits a signal to fibroblast-like synoviocytes from RA patients (RA-FLS). Methods: The induction of pro-inflammatory cytokines, chemokines, and mediators by sCD14 stimulation of RA-FLS was quantified by real-time PCR and ELISA. Cell proliferation was assessed by the BrdU assay. LPS-RS, a Toll-like receptor 4 (TLR-4) antagonist, was used to block TLR-4 signaling. Results: Soluble CD14 induced the expression of IL-6 mRNA and secretion of the protein. The expression of other pro-inflammatory cytokines and mediators, such as TNF-α, IL-8, intercellular adhesion molecule-1 (ICAM-1), MMP-3, and RANK ligand (RANKL), was also induced by sCD14. In addition, sCD14 stimulation promoted RA-FLS proliferation. LPS-RS abolished IL-6, IL-8, and ICAM-1 mRNA induction by sCD14 in RA-FLS. On the other hand, TNF-α and IL-17A increased TLR-4 expression by RA-FLS and amplified their sCD14-induced IL-6 expression. Conclusions: Soluble CD14 transmits inflammatory signals to RA-FLS via TLR-4. The effects of sCD14 may be augmented in inflammatory milieu. Our results suggest that sCD14 is involved in the pathogenesis of RA and may be a novel therapeutic target.

Association between yogurt consumption and plasma soluble CD14 in two prospective cohorts of US adults.

Although evidence suggests an inverse association between yogurt consumption and the risk of disorders, such as type 2 diabetes and certain cancers, the mechanisms remain poorly understood. We aimed to examine the association between yogurt consumption and concentrations of plasma soluble CD14, a marker of gut barrier dysfunction. We analyzed cross-sectional data from 632 women in the Nurses' Health Study (1989-1990) and 444 men in the Health Professionals Follow-up Study (1993-1994) with soluble CD14 concentrations. We estimated yogurt consumption from food frequency questionnaires. We used multivariable-adjusted linear regression models to estimate the percentage difference (95% CI) of soluble CD14 concentrations by yogurt consumption. Among men, higher consumption was associated with a lower soluble CD14 concentration (at least 2 cups/week vs. non-consumers; unadjusted % difference: -7.6%; 95% CI -13.0%, -2.1%; Ptrend = 0.003). The inverse association was slightly attenuated following multivariable adjustment (% difference: -5.8%; 95% CI -11.0%, -0.1%; Ptrend = 0.01). For the same comparison, yogurt consumption was inverse, but not statistically significant associated with soluble CD14 concentration in women (% difference: -1.2%; 95% CI -5.6%, 3.5%; Ptrend = 0.64). In stratified analyses, the inverse association between yogurt consumption and the concentrations of soluble CD14 was slightly stronger in men who consumed alcohol at least 20g/day. Higher yogurt consumption was associated with lower soluble CD14 concentrations, especially in men. Our findings suggest the strengthening of gut barrier function as a plausible mechanism for the observed inverse associations of yogurt consumption with gastrointestinal diseases and disorders involving other systems.

Impact of high-fat overfeeding on white adipose tissue and systemic metabolic and inflammatory responses

AbstractWhite adipose tissue (WAT) inflammation is linked to the development of cardiometabolic disease. A high intake of saturated fatty acids (SFA) may promote a more pro-inflammatory WAT gene expression profile, compared to monounsaturated fatty acids(1,2). This is concerning as the UK adult population currently exceeds the SFA intake target of &lt; 10% total energy (TE), at 11.9%TE(3). However, further research is needed to understand how SFA ingestion impacts the protein content and phosphorylation of key targets involved in WAT inflammatory signalling. Therefore, this study investigated the effect of short-term, high-fat overfeeding on WAT and systemic metabolic and inflammatory responses in healthy, non-obese individuals.Fifteen (9 men, 6 women) apparently healthy participants (mean (SE) age; 26.6 (1.2) y, BMI; 24.4 (0.6) kg/m2) consumed a high-fat (dietary target: 65.0%TE total fat; 32.5%TE SFA), high-calorie (+ 50% estimated energy requirements) diet for 7 days. Fasted venous blood and WAT samples were collected pre- and post-intervention.Body mass increased following the intervention (pre-intervention vs. post-intervention: 75.2 (2.9) kg vs. 76.4 (2.9) kg; P &lt; 0.0001). The intervention also led to increases in glucose (4.4 (0.1) vs. 4.6 (0.1) mmol/L; P = 0.022) and lipopolysaccharide-binding protein (12.9 (1.3) vs. 17.7 (3.0) μg/mL; P = 0.038), and a decrease in triacylglycerol (0.91 (0.08) vs. 0.72 (0.05) mmol/L; P = 0.007). Insulin, IL-6 and soluble CD14 concentrations remained unchanged (all P &gt; 0.05). At the WAT level, total protein (TLR4; NF-κB; IKβα; P38; MCP-1) and phosphorylation (p) of proteins (p-NF-κB; p-IKβα; p-P38) involved in inflammatory signalling were unchanged following the intervention (all P &gt; 0.05).In conclusion, although short-term, high-fat overfeeding led to an increase in body mass as well as systemic metabolic alterations, the content and phosphorylation of proteins involved in WAT inflammatory signalling were unaltered in healthy, non-obese individuals. Further analysis will reveal the impact of high-fat overfeeding on peripheral blood monocyte populations, which are involved in mediating pro- and anti-inflammatory responses.This research was supported by a Society for Endocrinology Early Career Grant.

Cluster of differentiation 14 and toll-like receptor 4 are involved in the anti-inflammatory effects of tyrosol

We investigated effects of tyrosol on upstream pathway regulating lipopolysaccharide (LPS)-induced inflammatory response, including cluster of differentiation 14 (CD14)-mediated LPS–macrophage binding, toll-like receptor 4 (TLR4)/myeloid differentiation factor 2 (MD2) expression and subsequent myeloid differentiation primary response gene 88 (MyD88) and TIR-domain-containing adapter-inducing interferon-β (TRIF) recruitments. Flow cytometry revealed that, compared with LPS-treated RAW264.7 cells (LPS group), those treated with LPS and tyrosol (1.2 mM) [LPS+T(1.2) group] demonstrated lower LPS–macrophage binding and membrane-bound CD14 (mCD14) and TLR4/MD2 expression (all p < 0.05). Immunoprecipitation/immunoblotting assay revealed lower MyD88 and TRIF concentrations in the LPS+T(1.2) group than in the LPS group (both p < 0.05). Soluble CD14 concentration was higher in the LPS+T(1.2) group than in the LPS group (p = 0.013); protease inhibition counteracted this effect. Thus, tyrosol inhibits LPS–macrophage binding, mCD14 and TLR4/MD2 expression, and MyD88 and TRIF recruitment, all possibly by enhancing protease-mediated mCD14 cleavage.

Structural Brain Abnormalities in Successfully Treated HIV Infection: Associations With Disease and Cerebrospinal Fluid Biomarkers.

Brain structural abnormalities have been reported in persons living with human immunodeficiency virus (HIV; PLWH) who are receiving suppressive combination antiretroviral therapy (cART), but their pathophysiology remains unclear. We investigated factors associated with brain tissue volumes and white matter microstructure (fractional anisotropy) in 134 PLWH receiving suppressive cART and 79 comparable HIV-negative controls, aged ≥45 years, from the Comorbidity in Relation to AIDS cohort, using multimodal neuroimaging and cerebrospinal fluid biomarkers. Compared with controls, PLWH had lower gray matter volumes (-13.7 mL; 95% confidence interval, -25.1 to -2.2) and fractional anisotropy (-0.0073; 95% confidence interval, -.012 to -.0024), with the largest differences observed in those with prior clinical AIDS. Hypertension and the soluble CD14 concentration in cerebrospinal fluid were associated with lower fractional anisotropy. These associations were independent of HIV serostatus (Pinteraction = .32 and Pinteraction = .59, respectively) and did not explain the greater abnormalities in brain structure in relation to HIV infection. The presence of lower gray matter volumes and more white matter microstructural abnormalities in well-treated PLWH partly reflect a combination of historical effects of AIDS, as well as the more general influence of systemic factors, such as hypertension and ongoing neuroinflammation. Additional mechanisms explaining the accentuation of brain structure abnormalities in treated HIV infection remain to be identified.

Relationship of salivary CD14 concentration with dental caries in young children

BackgroundDental caries is a chronic disease among children and there is need for studies assessing the caries risk factors using saliva. This study aimed to evaluate the association of salivary soluble CD14 and dental caries in young children.Material and MethodsA cross-sectional study was done among 300, 3-6 year old school children of Udupi district. A total of 40 children who were caries free, with no past systemic illness or craniofacial anomalies and 40 children with dental caries with no history of dental treatment for caries, with no past systemic illness or craniofacial anomalies were included in control and test groups respectively. Salivary CD14 was evaluated using ELISA test.ResultsThe mean salivary soluble CD14 concentration was significantly higher in caries free (1.34±0.35 µg/ml) children than caries experienced (0.54±0.36 µg/ml) (p<0.001). There was significant strong negative correlation between number of decayed teeth and soluble salivary CD14 (r = -0.868, P< 0.001) among all the children. Similarly, sub-group analysis of caries experienced children also showed significant strong negative correlation between number of decayed teeth and soluble salivary CD14 (r = -0.774, P<0.001).ConclusionsResults obtained in our study suggested that salivary CD14 can be a indicator of dental caries in young children. Key words:Caries, CD14, Children, Saliva.

The impact of vitamin D on the innate immune response to uropathogenic Escherichia coli during pregnancy

Urinary tract infections are highly common during pregnancy, and can cause serious complications for the mother and baby. Vitamin D, predominantly obtained from the sunlight, is known to have an effect on the urothelium, with immunomodulatory capacity against Escherichia coli infection. However, its influence at this site remains to be further explored. This study therefore investigated its impact during pregnancy in a population of women who have the possibility of adequate year-round sun exposure. Serum from pregnant Ugandan women (n = 32) in each trimester of pregnancy, from women after delivery (n = 29) and from never-pregnant controls (n = 25) was collected. 25-Hydroxyvitamin D (25-OHD), cathelicidin LL-37, human β-defensin 2, interleukin (IL)-8 and soluble CD14 serum concentrations were measured by chemiluminescence immunoassay or ELISA. The ability of serum to inhibit E. coli growth was tested. The immunomodulatory capacities of these serum samples and 1,25-dihydroxyvitamin D3 were investigated in urothelial cells. Increases in 25-OHD and LL-37 levels were observed as pregnancy progressed, peaking in the third trimester. Serum 25-OHD levels were higher in multigravidae than in primigravidae, and correlated positively with maternal age. IL-8 levels were lower in the third trimester than in the first trimester, increased after delivery, but remained below those of never-pregnant women. Similarly, soluble CD14 concentrations increased after delivery. As gestation advanced, serum had an increased capacity to inhibit E. coli growth. In vitro, it modulated the IL-8 response to infection in a vitamin D concentration-dependent manner. Our findings demonstrate that increasing vitamin D levels as pregnancy advances modulate the innate immune system towards a protective response to infection.

The role of CD14 gene promoter polymorphism in tuberculosis susceptibility

CD14 is expressed principally by cells of monocyte/macrophage lineage and plays a pivotal role in the innate immunity to intracellular infections. Recent research findings have revealed an association between the CD14 gene promoter polymorphism and several major infectious diseases. The aim of the present study was to investigate the association between the CD14-159C/T polymorphism and tuberculosis in a Turkish population. For this purpose, 88 consecutive patients with tuberculosis (63 pulmonary, 25 extrapulmonary) and 116 control subjects were enrolled into a prospective study. We determined CD14-159 genotypes by polymerase chain reaction - restriction fragment length polymorphism analysis and also measured serum concentrations of soluble CD14 (sCD14) by using a quantitative sandwich enzyme immunoassay technique. There was no significant difference in terms of genotype distribution between patients with tuberculosis (CC 18.2%, CT 48.9%, TT 33.0%) and controls (CC 12.9%, CT 50.9%, TT 36.2%) or between patients with pulmonary and extrapulmonary tuberculosis. Serum levels of sCD14 were significantly increased in patients with active tuberculosis compared to those with inactive tuberculosis and healthy controls (p<0.001). However, levels of sCD14 were not associated with any genotypes of CD14-159. The genotyping findings of the present study do not support a role for the CD14-159C/T polymorphism in the development of tuberculosis, at least in the geographical region of central Anatolia. Significantly elevated serum sCD14 levels in patients with active disease reflect the importance of the mononuclear phagocytic system activation in tuberculosis.

Analysis of Components of the CD14/TLR System on Leukocytes of Patients with Atopic Dermatitis

Background: Atopic dermatitis (AD) is frequently associated with skin infections that may be a consequence of an impaired function of the innate immune response. Conversely, the frequent bacterial colonization may also influence the systemic immune reactions, including the Toll-like receptor (TLR) system, through the translocation of bacterial components into the circulation. Therefore, we characterized phenotypic and functional properties of the TLR system in patients with extrinsic and intrinsic AD. Methods: The absolute number of surface CD14, TLR2, TLR4 and CD180 and the CD14-mediated uptake of bodipy-labeled endotoxin and bacteria by whole blood leukocytes was studied by flow cytometry. We measured the serum soluble CD14 concentration by an inhibitory flow cytometric method. Results: We observed a significant overexpression of TLR2 and TLR4 on monocytes, TLR2 and CD14 on granulocytes and CD180 on lymphocytes of intrinsic AD patients compared to healthy controls. The serum soluble CD14 was not different in the intrinsic AD patients, while it was diminished in the extrinsic AD group compared to the controls. The endotoxin and bacterium uptake showed no differences. Conclusions: The observed upregulation of CD14, TLR2, TLR4 and CD180 on peripheral leukocytes seems to be rather a consequence than the cause of the repeated bacterial infections in AD.

The effect of CD14-C159T genotypes on the cytokine response to endotoxin by peripheral blood mononuclear cells from asthmatic children

A C-T polymorphism at position 159 in the promoter of CD14 (C-159T) modulates the cellular response to endotoxin and significantly influences total IgE levels. The effect of this genetic variant on the cytokine response of the inflammatory cells is incompletely understood. To investigate the effects of CD14-C159T genotypes on the response to endotoxin by peripheral blood mononuclear cells (PBMCs) in children with asthma. The PBMCs from asthmatic children with the TT (n = 11) and CC (n = 11) genotypes at the CD14 promoter were cultured in the presence of endotoxin, 100 ng/mL; concanavalin A, 10 microg/mL; or medium alone. Concentrations of soluble CD14 (sCD14), interleukin (IL) 1beta, IL-4, IL-10, IL-12, IL-13, interferon-gamma, and transforming growth factor beta were determined in culture supernatants by enzyme-linked immunosorbent assay, and the transcriptional differences were evaluated using reverse-transcriptase polymerase chain reaction. Under unstimulated conditions, children with the TT genotype produced higher levels of sCD14 into the culture supernatant compared with children with the CC genotype (P = .03, Mann Whitney U test). Both IL-10 and IL-1beta concentrations were significantly higher in culture supernatants of children with the TT genotype after endotoxin stimulation (P = .02 and P = .009, respectively, by analysis of covariance [ANCOVA]). Messenger RNA expression was consistent with the results of protein concentration for IL-10 and sCD14. Concanavalin A stimulation resulted in lower levels of IL-4 in children with the TT genotype (P = .02, ANCOVA). The genotype at the CD14 promoter C159T locus may significantly influence the cytokine response of PBMCs obtained from asthmatic children. Differences in IL-10 and IL-4 production by alternative genotypes may contribute to the observed genotype effect on total IgE.

Breastfeeding, soluble CD14 concentration in breast milk and risk of atopic dermatitis and asthma in early childhood: birth cohort study

Breast milk contains a variety of bioactive substances, among them, soluble CD14 (sCD14), which plays an important role in innate immunity. We analysed data of a large prospective birth cohort study to examine the determinants of sCD14 in breast milk, and investigated whether breastfeeding practice and sCD14 concentrations in breast milk are determinants of the risk of atopic dermatitis (AD) and asthma in children. Eight hundred and three mothers and their newborns were included in this analysis. We measured sCD14 concentrations in breast milk samples collected 6 weeks post-partum. During a 2-year follow-up the cumulative incidences of AD and asthma were recorded. Overall, AD was reported for 20.6% of the 2-year-olds and asthma was reported for 19.6%. We found the lowest incidence of physician-reported AD in children of mothers without a history of atopic diseases if breastfed for 6 to less than 9 months. Furthermore, we found an inverse association between duration of breastfeeding and risk of asthma, which was especially evident in children with mothers without a history of atopic disease (P=0.01). These patterns persisted after control for other factors by multivariate analysis methods. The protective effect of breastfeeding seemed to be synergistic with sCD14 concentrations in breast milk (P for trend 0.0005). The results of this prospective birth cohort study suggest that a longer duration of breastfeeding does decrease the risk for asthma in early childhood, especially in children of mothers without a history of atopic disease. The beneficial effects of breastfeeding might be further supported by high levels of sCD14 in breast milk.

Peripheral blood monocyte activation during coronary artery bypass grafting with or without cardiopulmonary bypass

Objective. The aim of this prospective, randomized study was to investigate the impact of coronary artery bypass grafting (CABG) on peripheral monocytes and to evaluate the additional effect of cardiopulmonary bypass (CPB).Design. Twenty patients admitted for elective CABG were randomized to either on-pump (ONCAB, n=9) or off-pump (OFFCAB, n=11) surgery and blood samples were drawn before, during and 24 h after the operation. The total number of monocytes and the proportion of the more mature CD16+/CD14+ monocytes were measured. Expression of activation markers (CD11b, CD35 and CD62L) and oxidative burst were determined using flow cytometry on both resting and in vitro stimulated cells. Serum concentrations of soluble CD14 and monocytes/macrophage chemotactic protein 1 (MCP-1) were analysed.Results. During surgery there was a selective decrease in the proportion of CD16+/CD14+ monocytes compared to total monocytes. These had returned to preoperative values 24 h after surgery while the total number of monocytes had increased more than 100%. Intracellular production of oxygen free radical H2O2 was increased in the ONCAB group during surgery compared to OFFCAB. Monocyte expression and in vitro mobilization of complement receptors, CD11b and CD35, were similar in both study groups during and after surgery as was the expression of CD62L. Serum levels of MCP-1 decreased during surgery as did soluble CD14, both with increased levels again the day after surgery.Conclusion. It is concluded that the circulating monocyte population is activated during and as a consequence of CABG. There were few apparent additional effects of CPB found in this study. In this setting the inflammation caused by the surgery procedure per se probably surpasses the impact of the CPB on circulating blood monocytes.

Adhesion molecules as a prognostic marker of liver cirrhosis

ObjectiveEndothelial activation plays an active role in modifications of the circulatory status of cirrhotic patients. Soluble endothelial adhesion molecules, induced by pro-inflammatory cytokines, could be considered markers of endothelial activation. Their role in the natural history of cirrhosis and portal hypertension has not been reported. Our objective was to analyze the prognostic value of soluble adhesion molecules in cirrhotic patients.Material and methodsSerum concentrations of soluble CD14, soluble receptors of tumor necrosis factor alpha and adhesion molecules ICAM-1 (intercellular adhesion molecule-1) and VCAM-1 (vascular cell adhesion molecule 1) as well as mean blood pressure, plasma renin activity, aldosterone, vasopressin and norepinephrine concentrations were determined in 64 cirrhotic patients (Child Pugh class: A 48.4%, B 34.4%, C 17.2%), without any evidence of infection, and in 25 healthy controls. Patients were followed-up for a mean of 36.4 (range 6–60) months.ResultsIncreased concentrations of soluble CD14, tumor necrosis factor receptors and ICAM-1 and VCAM-1 were detected in cirrhotic patients when compared with healthy controls. Tumor necrosis factor receptors and adhesion molecule concentrations were both significantly higher in advanced phases of cirrhosis (Child Pugh class C and B versus A). Fifteen patients died as a related consequence of liver cirrhosis. Multivariate analysis demonstrated that Child-Pugh score and serum levels of tumor necrosis factor receptor I and ICAM-1 were associated with mortality.ConclusionsIn addition to the classic factor implicated in mortality (Child-Pugh class), alterations in inflammation-related components and soluble adhesion molecules, as representatives of hemodynamic alterations, are of prognostic significance in cirrhotic patients.

Opposite relationship between circulating soluble CD14 concentration and endothelial function in diabetic and nondiabetic subjects

Recent prospective studies indicate endothelial dysfunction and increased risk for cardiovascular events in patients with serological evidence of multiple infections. Soluble CD14 (sCD 14) plays a key role in the neutralization of lipopolysaccharide (LPS), a well-established bacterial product inducing endothelial dysfunction. Insulin resistance was recently identified as a significant factor influencing circulating sCD 14 concentration. Thus, we investigated the association of circulating sCD14 and endothelial dysfunction in subjects with well-established insulin resistance (patients with type 2 diabetes, n = 40) compared to control non-diabetic subjects (n = 100). To further explore the underlying mechanisms, we also analysed C-reactive protein and circulating NO2-/NO3- and cyclic GMP in the diabetic group. Serum sCD 14 concentration (ELISA) was found to be differently associated with endothelium-dependent vasodilatation (EDVD, high-resolution ultrasound) in diabetic and non-diabetic subjects. In nondiabetic subjects, serum sCD14 and C-reactive protein correlated negatively with EDVD (r = -0.21, p = 0.03, and r = -0.21, p = 0.03, respectively). In a partial correlation analysis, these associations remained significant after controlling for age and weight (sCD 14 and EDVD, r = -0.23, p = 0.023; C-reactive protein and EDVD, r = -0.21, p = 0.03; sCD14 and C-reactive protein, r = 0.30, p = 0.002). In contrast, sCD 14 was positively associated with EDVD in type 2 diabetic patients (r = 0.37, p = 0.019,). Interestingly, sCD14 was also associated with NO2-/NO3- in this group (r = 0.62, p = 0.001, n = 22). EDVD also correlated with cyclic GMP (r = 0.47, p = 0.03, n = 22). In summary, circulating sCD 14 is associated with endothelial function. While in non-diabetic subjects sCD14 behaves as an acute phase reactant, its role in type 2 diabetic patients should be further clarified. These findings need to be confirmed in further studies with larger number of patients.

Association of the -159 C --> T polymorphism in the CD14 promoter with variations in serum lipoproteins in healthy subjects.

The CD14-159 C --> T polymorphism, a single nucleotide polymorphism (SNP) at position -159 in the promoter region of the gene encoding the pattern recognition receptor CD14, has been associated with elevated plasma concentrations of soluble CD14, lowered serum immunoglobulin E, increased risk for myocardial infarction, and decreased risk for allergy and asthma. In the present study, the CD14-159 C --> T polymorphism has been investigated in order to determine its frequency and association with proinflammatory variables and lipid profile traits of 117 volunteers. The frequency of the CD14 promoter genotype as determined by polymerase chain reaction amplification-restriction fragment length polymorphism analysis was 35.0% (CC), 44.4% (CT), and 20.5% (TT), and the T allele frequency was 42.7%. Compared with the other genotypes, notably CC homozygotes, TT homozygotes were associated with lower total cholesterol, low-density lipoprotein cholesterol and apolipoprotein B-100 (P < 0.01) concentrations in serum. However, no association was found between the investigated SNP and inflammatory mediators such as fibrinogen, interleukin-6, tumor necrosis factor-alpha, tissue factor, C-reactive protein, plasminogen activator inhibitor-1, leukotriene B4, or thromboxane B2. In conclusion, the CD14-159 C --> T polymorphism may be an important genetic trait, related to the ability of CD14 to bind and transport lipids, such as cholesterol.

Tumour necrosis factor (TNF) gene polymorphism influences TNF-alpha production in lipopolysaccharide (LPS)-stimulated whole blood cell culture in healthy humans.

TNF-alpha is involved in infectious and immuno-inflammatory diseases. Different individuals may have different capacities for TNF-alpha production. This might determine a predisposition to develop some complications or phenotypes of these diseases. The aims of our study were to assess the inter-individual variability of TNF-alpha production and to correlate this variability to a single base pair polymorphism located at position -308 in TNF gene. We studied 62 healthy individuals. TNF-alpha production after LPS stimulation was evaluated using a whole blood cell culture model. The TNF gene polymorphism was studied by an allele-specific polymerase chain reaction. Other cytokines produced in the culture, soluble CD14 concentrations and expression of CD14 on blood cells were also measured. Among the 62 individuals, 57 were successfully genotyped. There were 41 TNF1 homozygotes and 16 TNF1/TNF2 heterozygotes. TNF-alpha production after LPS stimulation of whole blood cell culture was higher among TNF2 carriers than among TNFI homozygotes (929pg/ml (480-1473pg/ml) versus 521 pg/ ml (178-1307 pg/ml); P<0.05). This difference was even more significant after correction of TNF-alpha production for CD14 expression on blood cells. In conclusion, the single base pair polymorphism at position -308 in the TNF gene may influence TNF-alpha production in healthy individuals.