Cytokine Ratio Research Articles

Key serum cytokine markers for evaluating the efficacy of vaccine therapy in autoimmune models of rheumatoid arthritis

Abstract Background Pro-inflammatory T helper (Th)17 or Th1 cell responses (IL17, IFNγ, TNFα) drive autoimmune pathology in rheumatoid arthritis (RA) at the expense of anti-inflammatory/regulatory (IL4, IL10, TGFβ) T cell responses. Ligand Epitope Antigen Presenting System (LEAPS) vaccines, composed of cell-binding and immunodominant peptides, can down-modulate Th17 or Th1 activity and thus have the potential to treat ongoing RA. Choice of appropriate treatment requires knowledge of key cytokines driving disease as well as responses to therapy. Experimental design Serum cytokines were assayed in mouse models of RA including proteoglycan-induced arthritis (PGIA) recombinant proteoglycan G1 domain-induced arthritis (GIA) and collagen-induced arthritis (CIA) after arthritic mice had been treated with LEAPS vaccines. Serum cytokine levels and ratios of anti-inflammatory to pro-inflammatory cytokines were correlated with visual and histopathologic scores of disease. Results Serum levels of pro-inflammatory cytokines were elevated in control mice with either PGIA, GIA, or CIA. Reduced disease progression in mice treated with LEAPS vaccines was accompanied by decreased pro-inflammatory cytokine levels and increased ratios of anti-inflammmatory/regulatory to pro-inflammatory cytokines. Conclusion Serum concentrations and ratios of key disease-related cytokines can predict therapeutic efficacy in different animal models of RA. Successful vaccine therapy is associated with ratios shifted in favor of anti-inflammatory/regulatory cytokines. The ability to predict cytokine responses to therapy will allow better design or choice of appropriate immunomodulatory LEAPS vaccines and other therapies in RA.

Plasma cytokines associated with febrile status epilepticus in children: A potential biomarker for acute hippocampal injury.

Our aim was to explore the association between plasma cytokines and febrile status epilepticus (FSE) in children, as well as their potential as biomarkers of acute hippocampal injury. Analysis was performed on residual samples of children with FSE (n=33) aspart of the Consequences of Prolonged Febrile Seizures in Childhood study (FEBSTAT) and compared to children with fever (n=17). Magnetic resonance imaging (MRI) was obtained as part of FEBSTAT within 72h of FSE. Cytokine levels and ratios of antiinflammatory versus proinflammatory cytokines in children with and without hippocampal T2 hyperintensity were assessed as biomarkers of acute hippocampal injury after FSE. Levels of interleukin (IL)-8 and epidermal growth factor (EGF) were significantly elevated after FSE in comparison to controls. IL-1β levels trended higher and IL-1RA trended lower following FSE, but did not reach statistical significance. Children with FSE were found to have significantly lower ratios of IL-1RA/IL-1β and IL-1RA/IL-8. Specific levels of any one individual cytokine were not associated with FSE. However, lower ratios of IL-1RA/IL-1β, IL-1RA/1L-6, and IL-1RA/ IL-8 were all associated with FSE. IL-6 and IL-8 levels were significantly higher and ratios of IL-1RA/IL-6 and IL-1RA/IL-8 were significantly lower in children with T2 hippocampal hyperintensity on MRI after FSE in comparison to those without hippocampal signal abnormalities. Neither individual cytokine levels nor ratios of IL-1RA/IL-1β or IL-1RA/IL-8 were predictive of MRI changes. However, a lower ratio of IL-1RA/IL-6 was strongly predictive (odds ratio [OR] 21.5, 95% confidence interval [CI] 1.17-393) of hippocampal T2 hyperintensity after FSE. Our data support involvement of the IL-1 cytokine system, IL-6, and IL-8 in FSE in children. The identification of the IL-1RA/IL-6 ratio as a potential biomarker of acute hippocampal injury following FSE is the most significant finding. If replicated in another study, the IL-1RA/IL-6 ratio could represent a serologic biomarker that offers rapid identification of patients at risk for ultimately developing mesial temporal lobe epilepsy (MTLE).

Systemic pro-inflammatory cytokine status following therapeutic hypothermia in a piglet hypoxia-ischemia model

BackgroundInflammatory cytokines are implicated in the pathogenesis of perinatal hypoxia-ischemia (HI). The influence of hypothermia (HT) on cytokines after HI is unclear. Our aim was to assess in a piglet asphyxia model, under normothermic (NT) and HT conditions: (i) the evolution of serum cytokines over 48 h and (ii) cerebrospinal fluid (CSF) cytokine levels at 48 h; (iii) serum pro/anti-inflammatory cytokine profile over 48 h and (iv) relation between brain injury measured by magnetic resonance spectroscopy (MRS) and brain TUNEL positive cells with serum cytokines, serum pro/anti-inflammatory cytokines and CSF cytokines.MethodsNewborn piglets were randomized to NT (n = 5) or HT (n = 6) lasting 2–26 h after HI. Serum samples were obtained 4–6 h before, during and at 6–12 h intervals after HI; CSF was obtained at 48 h. Concentrations of interleukin (IL)-1β, −4, −6, −8, −10 and TNF-α were measured and pro/anti-inflammatory status compared between groups. White matter and thalamic voxel lactate/N-acetyl aspartate (Lac/NAA) (a measure of both oxidative metabolism and neuronal loss) were acquired at baseline, after HI and at 24 and 36 h.ResultsLac/NAA was reduced at 36 h with HT compared to NT (p = 0.013 basal ganglia and p = 0.033 white matter). HT showed lower serum TNF-α from baseline to 12 h (p < 0.05). Time-matched (acquired within 5 h of each other) serum cytokine and MRS showed correlations between Lac/NAA and serum IL-1β and IL-10 (all p < 0.01). The pro/anti-inflammatory ratios IL-1β/IL-10, IL-6/IL-10, IL-4/IL-10 and IL-8/IL-10 were similar in NT and HT groups until 36 h (24 h for IL-6/IL-10); after this, 36 h pro/anti-inflammatory cytokine ratios in the serum were higher in HT compared to NT (p < 0.05), indicating a pro-inflammatory cytokine surge after rewarming in the HT group. In the CSF at 48 h, IL-8 was lower in the HT group (p < 0.05). At 48 h, CSF TNF-α correlated with Lac/NAA (p = 0.02) and CSF IL-8 correlated with white matter TUNEL positive cell death (p = 0.04).ConclusionsFollowing cerebral HI, there was a systemic pro-inflammatory surge after rewarming in the HT group, which is counterintuitive to the putative neuroprotective effects of HT. While serum cytokines were variable, elevations in CSF inflammatory cytokines at 48 h were associated with MRS Lac/NAA and white matter cell death.

Association of self-reported symptoms with serum levels of vitamin D and multivariate cytokine profile in healthy women.

BackgroundAlthough a large number of studies have investigated possible relationships among serum levels of vitamin D or cytokines with disease progress and prognosis, similar studies on self-reported symptoms are still controversial. The overall objective of this study was to look into the association between serum levels of vitamin D or cytokines with self-reported symptoms related to musculoskeletal pain, sleep disorders, and premenstrual syndrome (PMS) in healthy adult women.Subjects and methodsVenous blood samples were collected from 117 healthy adult women, and serum levels of vitamin D, pro-inflammatory cytokines (IL-1β, IL-6, IL-8, IL-17, IFN-γ, and TNF-α) and anti-inflammatory cytokines (IL-4, IL-10, and IL-13) were measured. Groups were tested for differences in single parameters, pro-:anti-inflammatory cytokine ratios, and differences in multivariate patterns.ResultsThere were no significant associations between serum levels of vitamin D and any of the self-reported symptoms studied. However, serum levels of certain pro-inflammatory cytokines were significantly higher in subjects with musculoskeletal pain (IL-8, P=0.008), sleep disorders (IFN-γ, P=0.02), and PMS (IL-8 and TNF-α, P=0.009 and 0.002, respectively) compared to subjects who reported no symptoms. The pro-:anti-inflammatory cytokine ratios showed pro-inflammatory cytokine dominance in subjects with self-reported symptoms, particularly in the groups with deficient levels of vitamin D. However, the multivariate cytokine-pattern analysis was significantly different between PMS groups only.ConclusionThese data point to a possible role of pro-inflammatory cytokines as a contributing factor in self-reported symptoms related to musculoskeletal pain, sleep disorders, and PMS.

Peripheral inflammatory cytokines and immune balance in Generalised Anxiety Disorder: Case-controlled study

Introduction: Previous investigations have demonstrated that major depression is associated with particular patterns of cytokine signalling. The primary aim of this study was to examine peripheral pro-inflammatory and anti-inflammatory cytokines and immune balance in Generalised Anxiety Disorder (GAD). Methods: A case-controlled cross-sectional study design was employed: 54 patients with GAD and 64 healthy controls were recruited. Participants completed self-report measures of anxiety and depression. Two pro-inflammatory and two anti-inflammatory cytokines were measured using multiplex technology. Results: Case-control logistic regression analyses revealed significant differences in serum levels of IL-10, TNF-α, and IFN-γ between GAD and control groups after adjusting for age, gender, body mass index, smoking and alcohol consumption: these group differences were independent of the presence or degree of depression. Comparison of pro-inflammatory to anti-inflammatory cytokine ratios indicated that there were significantly higher ratios of TNF-α/IL10, TNF-α/IL4, IFN-γ/IL10, and IFN-γ/IL4 in the GAD group compared to the control group. Conclusions: This study is the first to investigate both pro- and anti-inflammatory cytokines and their balance in patients with GAD in comparison to healthy controls. The findings indicate a relatively increased pro-inflammatory response and decreased anti-inflammatory response and provide the first demonstration of an altered cytokine balance in GAD. Serum cytokine levels in GAD were independent of the presence of depression.

Cytokine profiles at birth and the risk of developing severe respiratory distress and chronic lung disease.

Background:Neonates with the diagnosis of respiratory distress syndrome (RDS) were studied to investigate possible associations between cytokine levels at birth and developing severe RDS or chronic lung disease (CLD).Materials and Methods:This was a cross-sectional study on serum and bronchoalveolar lavage (BAL) samples collected within hours of birth from infants with moderate and severe RDS. Twenty infants with moderate RDS and 20 infants with severe RDS were studied. RDS was diagnosed on the basis of radiographic findings, respiratory distress, and an increasing oxygen requirement. RDS severity was graded based on the radiological findings and Downe's Score. CLD was diagnosed when infants were still on supplemented O2 by at least 28 days of age. Levels of the cytokines interleukin (IL)-1β, IL-6, IL-8, IL-10, and tumor necrosis factor alpha were measured using enzyme-linked immunosorbent assay. “Statistical analysis was performed using the SPSS for Windows, (SPSS Inc., Chicago, IL, USA).”Results:Levels of the proinflammatory cytokines IL-8 and IL-1β were significantly higher in BAL of infants with severe RDS than those with moderate RDS (P = 0.007 and P = 0.02, respectively). IL-8 levels were also significantly higher in BAL and serum of infants who later progressed to CLD than in those who did not (P = 0.03 for both). The IL-8/IL-10 cytokine ratio was significantly higher in the BAL of severe RDS infants than in moderate RDS (P = 0.01) and in the serum of infants who progressed to CLD than in those who did not (P = 0.03).Conclusion:Levels of IL-8 and the IL-8/IL-10 ratio measured soon after birth were associated with severity of RDS as well as progression to CLD. Early measurement of cytokines levels and ratios may contribute to the prognosis and management of RDS and CLD.

Cytokine ratios in chronic periodontitis and type 2 diabetes mellitus

AimChronic periodontitis may influence systemic cytokines in type 2 diabetes. This study aimed to evaluate the cytokine ratios in type 2 diabetes with, and without chronic periodontitis. Materials & methodsGingival status, periodontal, glycemic parameters and serum cytokines were evaluated in participants grouped as healthy, chronic periodontitis, and type 2 diabetes with, and without chronic periodontitis. Results and conclusionsCytokine ratios showed significant differences in type 2 diabetes and chronic periodontitis, were highest in participants having both type 2 diabetes and chronic periodontitis, with a statistically significant cut-off point and area under curve by receiver operating characteristic.

Comprehensive Inflammatory Cytokine Profiling Identifies IL-8/CXCL8 As Elevated, Associated with Proliferative Features, and Independently Prognostic in Chronic Myelomonocytic Leukemia (CMML)

Introduction: Proinflammatory cytokines/chemokines have been implicated in the pathogenesis and prognosis of myeloproliferative neoplasms (MPNs) and myelodysplastic syndromes (MDS). Augmenting cytokine levels by JAK2 inhibition or cytokine neutralization are promising therapeutic approaches in these diseases. Chronic Myelomonocytic Leukemia (CMML) is a myeloid neoplasm that has both features of MDS and MPNs for which JAK2 inhibition has yielded reductions in inflammatory cytokines and promising clinical activity. Therefore, we hypothesize that annotation of inflammatory cytokines may uncover cytokine subsets associated with CMML-specific clinical features and identify novel therapeutic targets. To address this, we perform the first comprehensive annotation of inflammatory cytokines in CMML.Methods: A custom 46-plex Luminex based inflammatory cytokine profiling assay was developed. Cryopreserved peripheral blood plasma was collected from three academic centers after institutional review board approval. Center specific, age-matched plasma controls processed in a similar manner to CMML plasma samples were collected for analytical comparison. All samples were profiled in duplicate and only cytokines above the range of negative controls were reported. CMML cases were also clinically annotated by retrospective chart review and genotyping performed to annotate the frequency of TET2, IDH1/2, DNMT3A, ASXL1, EZH2, SRSF2, SF3B1, U2AF1, ZRSR2, N/K-RAS, CBL, and JAK2 somatic mutations. Descriptive statistics were used to summarize data and the Mann-Whitney test was used to compare cytokine levels among clinically and genetically relevant groups. A Cox proportional hazards model was used to derive likelihood ratios for overall survival using JMP®, Version 12. SAS Institute Inc., Cary, NC.Results: A total of 219 CMML and 35 control plasma samples were profiled. The median age was 73.4 years and 67.1 years in CMML and healthy controls, respectively. Eighty seven percent were CMML-1 by the WHO criteria and 54% were MPN-CMML by FAB criteria. Significantly different levels of cytokines between CMML and controls are summarized in Figure 1 and included TNFa (P=0.0404), IL-8 (P=0.0271), MCP-1 (P<0.0001), IL-1RA (P=0.0174), MIP-1a (P =0.0272), IL-17A (P= 0.0174), HGF (P =0.0006), and IL-12 (P=0.0287). Next, we analyzed differences in cytokine levels among clinically distinct groups to include proliferative versus dysplastic CMML patients reasoning that inflammatory cytokines are likely associated with this clinical phenotype. Indeed, patients with MPN-CMML and those with constitutional symptoms had increased levels of VEGF (P=0.0001), IL-10 (P=0.0002), IL-8 (P=0.0031), IL-1β (p=0.004), and other cytokines summarized in Figure 2 consistent with their clinical symptomatology. Last, we performed a genotype cytokine analysis and identified mutation-specific CMML cytokine signatures summarized in Figure 2. For example, patients with TET2 mutations were found to have significantly increased IFNγ (P=0.0470), GM-CSF (0.0020), and CD44 (P=0.058) while splicing mutations had significantly increased levels of MIP-1a (P=0.049), IL-2 (P=0.017), and CD44 (P=0.0039). To explore whether cytokines can independently influence prognosis, we used a cox proportional hazard model to calculate likelihood ratios of cytokines and overall survival. In univariate analysis, we identified VEGF (P=0.0006), IFNγ (P=0.001), EGF (P=0.0148), PDGF-CC (P=0.0165), and IL-10 (P=0.046) as prognostic in CMML (Figure 3). We next adjusted for age, hemoglobin, FAB and WHO subtype and demonstrate that only CXCL8 (p=0.016), PDGF-CC (P=0.010), and IL-10 (p=0.003) were independently prognostic.Conclusions: CMML is characterized by specific cytokine signatures that are associated with clinical disease behavior and somatic mutations. IL-8/CXCL8 was the only cytokine/chemokine in our analysis that was statistically elevated, associated with proliferative features, and independently prognostic suggesting a critical role in CMML pathogenesis. The therapeutic candidacy of augmenting differentially expressed cytokines will be the subject of future investigation. [Display omitted] [Display omitted] [Display omitted] DisclosuresKomrokji:Novartis: Consultancy, Speakers Bureau; Celgene: Membership on an entity's Board of Directors or advisory committees, Research Funding. Santini:Novartis: Consultancy, Honoraria; Amgen: Other: advisory board; Astex: Other: advisory board; Celgene: Consultancy, Honoraria, Research Funding; Onconova: Other: advisory board; Janssen: Consultancy, Honoraria.

Granulocyte colony-stimulating factor decreases the Th1/Th2 ratio in peripheral blood mononuclear cells from patients with chronic immune thrombocytopenic purpura in vitro

Chronic immune thrombocytopenia purpura (ITP) is an autoimmune disease that exhibits an abnormally high Th1/Th2 ratio. Granulocyte colony-stimulating factor (G-CSF) has been shown to decrease the Th1/Th2 ratio in healthy donors. In this study, we investigated the effects of G-CSF treatment on the Th1/Th2 cells and the underlying mechanisms in patients with ITP in vitro. Peripheral blood mononuclear cells (PBMCs) isolated from patients with ITP and healthy controls were treated with G-CSF. Expression levels of interferon (IFN)-γ, interleukin (IL)-2, IL-4, and IL-13 in supernatants were measured by enzyme-linked immunosorbent assays. The expression of IFN-γ, IL-4, and G-CSF receptor (G-CSFR) on Th1 and Th2 cells were examined by flow cytometry and confocal microscopy. The mRNA expression of IFN-γ, IL-2, IL-4, IL-13, and T-box expressed in T cells (T-bet) and GATA-binding protein 3 (GATA-3) in PBMCs was evaluated by reverse transcription polymerase chain reaction. The results showed that G-CSF could significantly reduce the Th1/Th2 ratio in PBMCs from patients with ITP in vitro. As the concentration of G-CSF increased, Th1/Th2 ([IFN-γ+IL-2]/[IL-4+IL-13]) cytokine ratios and T-bet/GATA-3 mRNA ratios decreased in a concentration-dependent manner. Th1 cells and Th2 cells both expressed G-CSFR. These results suggest that G-CSF could decrease the Th1/Th2 ratio in the context of ITP, and elucidate the direct and indirect immunomodulatory mechanisms underlying G-CSF functions in Th1/Th2 cells, thus supporting the therapeutic potential of G-CSF in the treatment of patients with ITP.

TNF-α/IL-10 Ratio Correlates with Burn Severity and May Serve as a Risk Predictor of Increased Susceptibility to Infections.

Severe burn injury renders patients susceptible to multiple infection episodes; however, identifying specific patient groups at high risk remains challenging. Burn-induced inflammatory response dramatically modifies the levels of various cytokines. Whether these changes could predict susceptibility to infections remains unknown. The aim of this study was to determine the early changes in the pro- to anti-inflammatory cytokine ratio and investigate its ability to predict susceptibility to repeated infections after severe burn trauma. The patient population consisted of 34 adult patients having early (≤48 h since injury) blood draws following severe (≥20% total burn surface area (TBSA)) burn injury and suffering from a first infection episode at least 1 day after blood collection. Plasma TNF-α and IL-10 levels were measured to explore the association between the TNF-α/IL-10 ratio, hypersusceptibility to infections, burn size (TBSA), and common severity scores (Acute Physiology and Chronic Health Evaluation II (APACHEII), Baux, modified Baux (R-Baux), Ryan Score, and Abbreviated Burn Severity Index (ABSI)). TNF-α/IL10 plasma ratio measured shortly after burn trauma was inversely correlated with burn size and the injury severity scores investigated, and was predictive of repeated infections (≥3 infection episodes) outcome (AUROC [95%CI] of 0.80 [0.63–0.93]). Early measures of circulating TNF-α/IL10 ratio may be a previously unidentified biomarker associated with burn injury severity and predictive of the risk of hypersusceptibility to repeated infections.

Abstract # 1866 How is the immune balance affected in patients with generalised anxiety disorder?

Introduction Greater understanding of the role of cytokines in interactions between the nervous and immune systems has led to integrative and explanatory models for neuropsychiatric disorders. Our study aimed to examine peripheral inflammatory cytokines and immune balance in generalised anxiety disorder (GAD). Methods A case-controlled cross-sectional study investigated associations between psychological and biological variables. 54 patients with GAD and 64 healthy controls were recruited. Participants completed self-report measures of anxiety and depression. A panel of two pro-inflammatory and two anti-inflammatory cytokines were measured using multiplex technology. Results Case-control logistic regression analyses revealed significant differences in serum levels of IL-10, TNF- α , and IFN- γ between GAD and control groups after adjusting for age, gender and body mass index: these group differences were independent of the presence or degree of depression. Comparison of pro- and anti-inflammatory cytokine ratios indicated that there were significantly higher ratios of TNF- α /IL10, TNF- α /IL4, IFN- γ /IL10, and IFN- γ /IL4 in the GAD group compared to the control group. Conclusions This study is the first to investigate both pro- and anti-inflammatory cytokines and their balance in patients with GAD in comparison to healthy controls. The findings indicate a relatively increased pro-inflammatory response and diminished anti-inflammatory response and provide the first demonstration of a Th1/Th2 cytokine imbalance in GAD. The changes in serum cytokine levels in GAD were independent of the presence of depression.

Impact of prior statin therapy on evaluation of the inflammatory process and cortisol concentration in patients with the first ST-segment elevation myocardial infarction undergoing coronary angioplasty with bare metal stent implantation

Introduction. The aim of the current study was to verify the impact of statin therapy on evaluation of the inflammatory process and cortisol concentration in patients with the first episode of ST-segment elevation myocardial infarction undergoing coronary angioplasty with bare metal stent implantation. Material and methods. The study was designed as a prospective, single-center cohort study, with a total number of 200 patients enrolled between 2005 and 2008. Results. Seventeen patients with ongoing statin therapy and 167 patients without those agents underwent pPCI with BMS implantation. The cortisol concentration on admission was significantly lower in statin therapy subgroup — median 14.9 μg/dL (9.5–23.3 IQR) vs. 28,1 μg/dL (17.2–39.9 IQR) for patients without lipid lowering drug, p = 0.01. The level of cortisol after 23h was also significantly lower in statin receiving subgroup: 10.4 μg/dL (8.3–19 IQR) vs. 17.3 μg/dL (10.1–25.9 IQR) adequately, p = 0.021. Those two groups did not differ significantly in relation to CRP, IL-6, IL-10 concentrations and to anti-inflammatory cytokine ratio [IL-6/IL-10] on admission, after 24h and at discharge. Significant differences were observed in the concentrations of cTnI and activities of CK and CK-MB. Conclusions. Ongoing statin therapy before an episode of myocardial infarction has an influence on cortisol concentration and cardiac markers. No significant effect on serum levels of cytokines has been observed.

Immune Cell-Conditioned Media Suppress Prostate Cancer PC-3 Cell Growth Correlating With Decreased Proinflammatory/Anti-inflammatory Cytokine Ratios in the Media Using 5 Selected Crude Polysaccharides.

Five different crude polysaccharides from guava seed (GSPS), bitter buckwheat (BBPS), common buckwheat (CBPS), red Formosa lambsquarters (RFLPS), and yellow Formosa lambsquarters (YFLPS) were isolated to treat human prostate cancer PC-3 cells via direct action or tumor immunotherapy. The splenocyte- and macrophage-conditioned media (SCM and MCM) were prepared using individual selected polysaccharides, and then SCM or MCM was further collected to treat PC-3 cells. The relationship between PC-3 cell growth and Th1/Th2 cytokines in SCM as well as proinflammatory/anti-inflammatory cytokine secretion profiles in MCM were delineated. The results showed that all 5 selected polysaccharides did not significantly inhibit PC-3 cell growth via direct action. However, SCM or MCM cultured in the absence or presence of 5 selected polysaccharides significantly (P < .05) inhibited PC-3 cell growth. MCM cultured with 5 polysaccharides dose dependently enhanced their inhibitory effects on the viabilities of PC-3 cells than those cultured without polysaccharides. There was a significant (P < .05) negative correlation between PC-3 cell viabilities and (interleukin [IL]-6 + tumor necrosis factor [TNF]-α)/IL-10 level ratios in the corresponding MCM, implying that macrophages suppress PC-3 cell growth through decreasing secretion ratios of proinflammatory/anti-inflammatory cytokines in a tumor microenvironment.

Beta-blockers decreases Th17/Treg cytokine ratios by down regulation of GRK2 synthesis in lymphocytes of patients with ACS

Objective To observe the effects of g protein-coupled receptor kinase-2 (GRK2) in peripheral lymphocytes on Th17/Treg cytokine ratios and investigate the potential impact and mechanism of beta-blockers on immune function in patients with acute coronary syndrome (ACS). Methods One hundred-four patients with ACS and forty-five normal coronary angiography (NCA) hospitalized patients as control group were enrolled in the study. ACS group was randomly divided into metoprolol group(n=52) and carvedilol group(n=52). The levels of lymphocyte GRK2 were detected by means of a sensitive sandwich enzyme-linked immunosorbent assay (ELISA) and IL-17 and FoxP3 in lymphocytes using flow cytometry at baseline and 6 months after the treatment. Results The levels of lymphocyte GRK2 and Th17/Treg in ACS patients were significantly higher than those of the UCA controls (bothP 0.05). Conclusion These results suggest that the increase of GRK2 may affect GRK2 in peripheral lymphocytes of ACS. Beta-blockers appear to have a beneficial effect on the Th17/Treg balance of immune system through decreasing activities of GRK2. Key words: Acute coronary syndrome; G protein-coupled receptor kinase-2; β-blockers; Th17/Treg

Serum and exhaled breath condensate inflammatory cytokines in community-acquired pneumonia: a prospective cohort study.

BackgroundThe role and relationship between pro- and anti-inflammatory cytokines represents one of the least studied aspects of the pathogenesis of community-acquired pneumonia (CAP). The aim of the present study was to evaluate pro- and anti-inflammatory cytokines at both local (lung) and systemic (blood) levels and their relationship with the severity of the disease on admission and time for a patient to reach clinical stability during hospitalisation.MethodsThis was an observational, prospective, cohort study of hospitalised patients with a diagnosis of CAP at the IRCCS Policlinico Hospital, Milan, Italy, between April 2010 and January 2012. Ten pro-inflammatory cytokines (interleukin [IL]-1, IL-1α, IL-1β, IL-2, IL-6, IL-8, tumor necrosis factor [TNF]α and interferon [IFN]γ) and anti-inflammatory cytokines (IL-4 and IL-10) were measured in both serum and exhaled breath condensate within 24 h after hospital admission.ResultsA total of 74 patients (median age: 76 years; gender: 61 % male) were enrolled. The anti- to pro-inflammatory cytokine ratio was reduced in patients with severe disease on admission and prolonged time to reach clinical stability. This was due to lower levels of anti-inflammatory cytokines in the exhaled breath condensate and higher levels of pro-inflammatory cytokines in serum.ConclusionsDis-regulation between pro- and anti-inflammatory pathways might be a part of the pathogenic mechanisms that lead to severe infection and worse early clinical outcomes in CAP patients.Electronic supplementary materialThe online version of this article (doi:10.1186/s41479-016-0009-7) contains supplementary material, which is available to authorized users.

Heavy metal mediated innate immune responses of the Indian green frog, Euphlyctis hexadactylus (Anura: Ranidae): Cellular profiles and associated Th1 skewed cytokine response

Immune cell and cytokine profiles in relation to metal exposure though much studied in mammals has not been adequately investigated in amphibians, due mainly to lack of suitable reagents for cytokine profiling in non-model species. However, interspecies cross reactivity of cytokines permitted us to assay levels of IFNγ, TNFα, IL6 and IL10in a common anuran, the Indian green frog (Euphlyctis hexadactylus), exposed to heavy metals (Cd, Cr, Cu, Zn and Pb, at ~5ppm each) under field and laboratory settings in Sri Lanka. Enumeration of immune cells in blood and melanomacrophages in the liver, assay of serum and hepatic cytokines, and Th1/Th2 cytokine polarisation were investigated.Immune cell counts indicated overall immunosuppression with decreasing total WBC and splenocyte counts while neutrophil/lymphocyte ratio increased with metal exposure, indicating metal mediated stress. Serum IL6 levels of metal exposed frogs reported the highest (~9360pg/mL) of all cytokines tested. Significantly elevated IFNγ production (P<0.05) was evident in heavy metal exposed frogs. Th1/Th2 cytokine ratio in both serum and liver tissue homogenates was Th1 skewed due to significantly higher production of pro-inflammatory cytokines, IFNγ in serum and TNFα in the liver (P<0.01).Metal mediated aggregations of melanomacrophages in the liver were positively and significantly (P<0.05) correlated with the hepatic expression of TNFα, IL6 and IL10 activity. Overall, Th1 skewed response may well be due to oxidative stress mediated nuclear factor κ-light chain enhancer of activated B cells (NFκB) which enhances the transcription of pro-inflammatory cytokines. Xenobiotic stress has recently imposed an unprecedented level of threat to wildlife, particularly to sensitive species such as amphibians. Therefore, understanding the interactions between physiological stress and related immune responses is fundamental to conserve these environmental sentinels in the face of emerging eco-challenges.

Correction: Immune Parameters That Distinguish Multiple Sclerosis Patients from Patients with Other Neurological Disorders at Presentation

Background/Aim Multiple sclerosis (MS) is an inflammatory, demyelinating disease of the central nervous system. Effector T helper cells, mainly Th1 and Th17, cytotoxic T-cells, B-cells, macrophages, microglia, and the cytokines they secrete, are implicated in the initiation and maintenance of a deregulated immune response to myelin antigens and the ensuing immune-mediated demyelination. In this study, we investigated whether signature cytokines exist in MS patients at presentation to gain an insight into the underlying immunopathogenic processes at the early stage of the disease. Methods We collected serum and cerebrospinal fluid (CSF) samples from 123 patients at presentation, eventually diagnosed with MS or non-inflammatory (NIND) or inflammatory neurological diseases (IND) or symptomatic controls (SC). The levels of cytokines IFN-γ, TNF-α, TGF-β1, IL-2, IL-4, IL-6, IL-10 and IL-17 were measured, and cytokine ratios, such as Th1/Th2, Th1/Th17, and Type-1/Type-2, were calculated. All parameters were tested for their correlations with the intrathecal IgG synthesis. Results Cytokine levels in CSF were lower than in serum in all the patients, with the exception of IL-6. Serum or CSF cytokine levels of MS patients did not differ significantly from NIND or SC, with the exception of serum IFN-γ and TNF-α that were significantly higher in NIND. IND patients presented with the highest levels of all cytokines in serum and CSF, with the exception of serum IL-10 and CSF IL-17. MS patients had a significantly lower serum Th1/Th2 ratio compared to the NIND and IND groups, and significantly lower serum Type-1/Type-2, IFN-γ/IL-10 and CSF Th1/Th17 ratios compared to IND patients. MS patients had a significantly higher CSF IL-17/IL-10 ratio compared to IND patients. The IgG index was higher in MS patients compared to the control groups; the differences reached statistical significance between the MS and the NIND and SC groups. Reiber-Felgenhauer analysis of the QIgG and QAlb indices revealed higher intrathecal IgG synthesis in MS patients, and higher blood-CSF barrier dysfunction in IND patients. The IgG index correlated with CSF IL-4 in MS patients only. Conclusions We found no signature cytokines or profiles thereof in MS patients at presentation. Only IND patients presented with a clear Th1 cytokine polarization in serum and CSF. The parameters that distinguished MS patients from patients with other neurological disorders were IgG intrathecal synthesis, the IgG index and its correlation with CSF IL-4 levels.

Aggression as an independent entity even in psychosis- the role of inflammatory cytokines

BackgroundAggression is very common in psychosis (prevalence ranging from 34% to 70%) and is often the main or first symptom for which the patient receives medical attention. Studies have associated alteration in cytokine profiles among healthy persons with aggressive traits. We hypothesise that even among those with psychosis, aggression is an independent entity, irrespective of psychotic state and is associated with cytokine alterations. To our knowledge, this is the first study attempting to look at the inflammatory cytokines in aggressive psychotic patients.Methods: Study included 80 participants divided into four groups viz. aggressive diseased, non aggressive diseased, aggressive non diseased and non aggressive non diseased depending upon presence or absence of aggression and psychosis. Interferon gamma(IFN-G), Interleukin 10(IL10) plasma concentrations and their ratio were measured using ELISA based assay kits read at absorbance of 450nm wavelength using Double beam spectrophotometer. The four groups were compared on measures of aggression, psychosis, Interferon Gamma levels, Interleukin 10 levels, Proinflammatory: Antiinflammatory cytokine ratio using standard statistical instruments. ResultsIn patients with psychosis, the cytokines IFN-G and IL10 were significantly lower compared to those without. The cytokines IFN-G and IL10 are both significantly associated both with aggression and psychosis. IL10, but not IFN-G is associated with aggression in absence of psychosis. The proinflammatory: antiinflammatory cytokine ratio, is more significantly associated with aggression, irrespective of psychosis. In fact, there is no significant relationship between the above ratio and psychosis. Strong correlation exists between the proinflammatory: antiinflammatory cytokine ratio and aggression scores, even after controlling for severity of psychosis. ConclusionsIt may be concluded from this study that in spite of a high prevalence of aggression in patients of psychosis, it is more likely to be an independent entity associated with individual cytokine changes and increased proinflammatory: antiinflammatory cytokine ratio as its hallmark.

Expression of M-CSF, TNF-β, IFN-γ, and IL-10 in Rats with Liver Cirrhosis and Hypersplenism and its Significance

Objective: This study aimed to examine the expression of M-CSF, TNF-β, IFN-γ, and IL-10 in rats with liver cirrhosis and hypersplenism, and to investigate its significance. Methods: Seventy-two healthy male SD rats were randomly divided into a model group (n=60) and a control group (n=12). The rats in the model group were first gavaged with a 40% CCL4/peanut oil solution to develop cirrhosis and hypersplenism. The expression rates and intensities of the pro-inflammatory cytokines M-CSF, TNF-β, and IFN-γ, and the anti-inflammatory cytokine IL-10 in the spleen were measured and compared with the control group. The pro-inflammatory/anti-inflammatory cytokine ratio and its role were analyzed. Results: The positive expression rates of M-CSF, TNF-β, IFN-γ, and IL-10 in the model group (cirrhosis and hypersplenism) were 61.76%, 79.41%, 64.70%, and 88.24%, respectively, which were significantly different (P<0.05) from the 25%, 33.33%, 16.67%, and 50% in the control group. The relative protein expression intensities of M-CSF, TNF-β, IFN-γ, and IL-10 in the model group were 0.63 ± 0.58, 1.06 ± 0.49, 0.99 ± 0.38, and 1.12 ± 0.42, respectively, and were significantly different (P<0.05) from the 0.18 ± 0.12, 0.52 ± 0.27, 0.38 ± 0.28, and 0.60 ± 0.32 in the control group. The relative mRNA expression levels of M-CSF, TNF-β, IFN-γ, and IL-10 in the model group were 2.06 ± 0.11, 4.07 ± 0.19, 2.98 ± 0.11, and 7.94 ± 0.27, respectively, and were significantly different (P<0.05) from the 1.01 ± 0.05, 1.06 ± 0.11, 1.00 ± 0.31, and 1.02 ± 0.08 in the control group. Conclusion: The abnormally increased expression of M-CSF, TNF-β, IFN-γ, and IL-10 in rats with liver cirrhosis and hypersplenism, as well as the abnormal pro-inflammatory/anti-inflammatory cytokine ratio and regulation may play an important role in enhancing the phagocytosis of macrophages and causing peripheral cytopenias.

The Effectiveness of Ablative Fractional Carbon Dioxide Laser with Autologous Platelet Rich Plasma Combined Resurfacing for Hypertrophic Scar of the Shoulder

Scar formation following surgery or trauma is common, and some scars gradually become reddish, hard, or hypertrophic. Scars are classified as hypertrophic, keloid, atrophic, or acne scars. Hypertrophic scars are firm and protrude due to increased collagen expression, but unlike keloid scars they do not extend beyond the margin of the original wound [1]. Laser treatment for scars has improved over the past three decades and provides a less invasive, less painful treatment option [1]. The most recent cutaneous laser technique is fractional photothermolysis, which thermally alters a “fraction” of the skin, leaving untouched areas of normal skin that rapidly repopulate the ablated columns of tissue. To date, there are few studies regarding the effectiveness of ablative fractional carbon dioxide (CO2) laser for treating hypertrophic scars. Platelets contain numerous granules and vast number of bioactive molecules that can facilitate homeostasis and wound healing. For these reasons, platelet-rich plasma (PRP) is increasingly being applied to clinical cases [1]. PRP is concentrated by centrifuging whole blood and then extracting a small volume of residual plasma. Autologous PRP is immunologically inert and contains the proper ratio of growth factors and cytokines [2]. Furthermore, additional equipment is not required to store PRP, and the processing cost is relatively low. Here we describe the outcomes of combining fractional CO2 laser therapy in combination with PRP to reduce a hypertrophic scar.