Marker Of Immune System Activation Research Articles

Novel Probable Glance at Inflammatory Scenario Development in Autistic Pathology.

Autism Spectrum Disorder (ASD) is characterized by persistent deficits in social communication and restricted-repetitive patterns of behavior, interests, or activities. ASD is generally associated with chronic inflammatory states, which are linked to immune system dysfunction and/or hyperactivation. The latter might be considered as one of the factors damaging neuronal cells. Several cell types trigger and sustain such neuroinflammation. In this study, we traced different markers of immune system activation on both cellular (immune cell phenotypes) and mediatory levels (production of cytokines) alongside adverse hematology and biochemistry screening in a group of autistic children. In addition, we analyzed the main metabolic pathways potentially involved in ASD development: energy (citric acid cycle components), porphyrin, and neurotransmitter metabolism. Several ASD etiological factors, like heavy metal intoxication, and risk factors—genetic polymorphisms of the relevant neurotransmitters and vitamin D receptors—were also analyzed. Finally, broad linear regression analysis allowed us to elucidate the possible scenario that led to the development of chronic inflammation in ASD patients. Obtained data showed elevated levels of urinary cis-aconitate, isocitrate, alfa-ketoglutarate, and HMG. There were no changes in levels of metabolites of monoamine neurotransmitters, however, the liver-specific tryptophan kinurenine pathway metabolites showed increased levels of quinolinate (QUIN) and picolinate, whereas the level of kynurenate remained unchanged. Abovementioned data demonstrate the infringement in energy metabolism. We found elevated levels of lead in red blood cells, as well as altered porphyrin metabolism, which support the etiological role of heavy metal intoxication in ASD. Lead intoxication, the effect of which is intensified by a mutation of the VDR-Taq and MAO-A, leads to quinolinic acid increase, resulting in energy metabolism depletion and mitochondrial dysfunction. Moreover, our data backing the CD4+CD3+ T-cell dependence of mitochondrial dysfunction development in ASD patients reported in our previous study leads us to the conclusion that redox-immune cross-talk is considered a main functional cell damaging factor in ASD patients.

Difficulties in psychosocial functioning due to current depressive symptoms: What can C-Reactive protein tell us?

BackgroundMultiple empirical studies and meta-analyses have examined how inflammation may be associated with various aspects of major depression, with older adults being particularly at risk for the effects of inflammation-related depression. Despite this wide area of research, no study has examined how depression-related inflammation impacts psychosocial functioning. MethodsData from the National Health and Nutrition Examination Survey, years 2007–2008, were utilized to examine whether adults over the age of 40 experienced difficulty in their work, taking care of things at home, or getting along with other people due to current depressive symptoms through a logistic regression analysis. We selected C-reactive protein (CRP), a common marker of immune system activation, as our primary predictor of interest while controlling for relevant covariates. ResultsGreater CRP was positively associated with a greater risk for individuals experiencing difficulties in psychosocial functioning due to depressive symptoms. While current number and severity of depressive symptoms was also found to be significant in the model, comparison of effect sizes identified that CRP appears to be a more relevant marker for experiencing difficulty than a number of relevant biopsychosocial covariates. ConclusionInflammation as measured by CRP may be a helpful tool in understanding how depressive symptoms are associated with an individual's ability to successfully navigate their social environment. Results here demonstrate the emerging utility of CRP in helping to assess the risk for negative outcomes in those experiencing depressive symptoms, especially as it pertains to older adults.

Balloon cells promote immune system activation in focal cortical dysplasia type 2b.

AimsFocal cortical dysplasia (FCD) type 2 is an epileptogenic malformation of the neocortex associated with somatic mutations in the mammalian target of rapamycin (mTOR) pathway. Histopathologically, FCD 2 is subdivided into FCD 2a and FCD 2b, the only discriminator being the presence of balloon cells (BCs) in FCD 2b. While pro‐epileptogenic immune system activation and inflammatory responses are commonly detected in both subtypes, it is unknown what contextual role BCs play.MethodsThe present study employed RNA sequencing of surgically resected brain tissue from FCD 2a (n = 11) and FCD 2b (n = 20) patients compared to autopsy control (n = 9) focusing on three immune system processes: adaptive immunity, innate immunity and cytokine production. This analysis was followed by immunohistochemistry on a clinically well‐characterised FCD 2 cohort.ResultsDifferential expression analysis revealed stronger expression of components of innate immunity, adaptive immunity and cytokine production in FCD 2b than in FCD 2a, particularly complement activation and antigen presentation. Immunohistochemical analysis confirmed these findings, with strong expression of leukocyte antigen I and II in FCD 2b as compared to FCD 2a. Moreover, T‐lymphocyte tissue infiltration was elevated in FCD 2b. Expression of markers of immune system activation in FCD 2b was concentrated in subcortical white matter. Lastly, antigen presentation was strongly correlated with BC load in FCD 2b lesions.ConclusionWe conclude that, next to mutation‐driven mTOR activation and seizure activity, BCs are crucial drivers of inflammation in FCD 2b. Our findings indicate that therapies targeting inflammation may be beneficial in FCD 2b.

The Influence of FLT3-ITD Mutation on Baseline Levels of Selected Cytokines and Soluble Adhesion Molecules in Acute Myeloid Leukemia with Mutated NPM-1

Introduction: Cytokines and adhesion molecules have been studied as markers of immune system activation in various diseases including AML/MDS. These factors form a dynamic network which affects AML cell susceptibility to chemotherapy. We analyzed whether these factors may be associated with mutations specific for AML. The aim of our study was to evaluate hypothesis that baseline serum levels of these factors may be associated with mutation profile on the model of AML with mutation in NPM-1 and FLT3-ITD. Methods: A total of 42 de novo AML patients (age 55.9 ± 11.5, median 60 years; 16 males, 26 females; all Caucasian) were analyzed. All patients had normal karyotype and mutation in NPM-1, 25 patients had mutation in FLT3-ITD. The subgroups have not differed in age. We evaluated baseline circulating levels of the following factors: interleukins (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-8, IL-10), epidermal growth factor (EGF), interferon-gamma (IFN-γ), monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-alpha (TNF-α), vascular endothelial growth factor (VEGF), E-selectin (E-SEL), P-selectin (P-SEL), L-selectin (L-SEL), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). All biomarkers were measured by biochip array technology on Evidence Investigator analyzer (Randox). Probability values (P) < 0.05 were considered statistically significant. Results: The cases with FLT3-ITD mutation had higher levels of E-SEL (51.90 ± 19.90 mcg/L vs. 23.85 ± 14.07 mcg/L; p = 0.002), P-SEL (192.01 ± 77.29 mcg/L vs. 127.89 ± 48.47 mcg/L; p = 0.035) and lower levels of EGF (7.01 ± 7.08 ng/L vs. 20.64 ± 16.73 ng/L; p = 0.019) and TNF-α (3.55 ± 1.89 ng/L vs. 5.69 ± 3.55 ng/L; p = 0.049). The levels of L-SEL were higher in FLT3-ITD mutated AML, which was not statistically significant, possibly due to exceeding the upper limit or array sensitivity in majority of cases. The differences in baseline levels of other evaluated markers were not statistically significant. Conclusions: Cytokines are an important part of cancer environment. In patients with normal karyotype and mutation in NPM-1, the presence of FLT3-ITD influenced levels of EGF, TNF-α, E-SEL, P-SEL and possibly L-SEL. Whether these findings are associated with adverse biology of FLT3-ITD positive leukemia is not known at the moment and requires further elucidation. Disclosures No relevant conflicts of interest to declare.

PS1048 BASELINE SERUM LEVELS OF RECEPTORS FOR INTERLEUKIN‐2 AND TUMOR NECROSIS FACTOR‐α PREDICT OVERALL SURVIVAL IN NEWLY DIAGNOSED ACUTE MYELOID LEUKEMIA

Background:Cytokines and adhesion molecules have been studied as markers of immune system activation in many diseases including acute myeloid leukemia (AML). Further knowledge gained from baseline cytokine levels assessment may help to improve risk stratification and outcomes.Aims:The aim of this study is to evaluate baseline serum levels of selected leukemic environment markers, search for relationship with progression‐free survival (PFS), overall survival (OS) and compare the findings with standard prognostic markers in AML.Methods:A total of 80 AML patients, age 53.5 ± 12.6 years, were studied in the period 2010 – 2016. The data were collected to the end of year 2018, so that all living event‐free patients had at least two‐year follow‐up. Patients with acute promyelocytic leukemia or ineligible for induction therapy were not included into the study. Risk stratification was carried out according to the ELN 2017 recommendations. All patients were induced with “3+7” chemotherapy. We evaluated baseline serum levels of interleukins (IL‐1α, IL‐1β, IL‐2, IL‐3, IL‐4, IL‐5, IL‐6, IL‐7, IL‐8, IL‐10, IL‐12, IL‐13, IL‐15), Epidermal Growth Factor, GM‐CSF, Interferon‐γ, Macrophage Inflammatory Protein‐1α, Monocyte Chemotactic Protein‐1, Tumor Necrosis Factor‐α, Vascular Endothelial Growth Factor, E‐selectin, P‐selectin, L‐selectin, Intercellular Adhesion Molecule‐1, Vascular Cell Adhesion Molecule‐1, Matrix Metalloproteinase‐9, soluble IL‐2 receptor‐α (sIL‐2Rα) and soluble receptors for IL‐6 (sIL‐6R) and TNF‐α type I and II (TNFR‐1,2). All biomarkers were measured by biochip array technology on Evidence Investigator analyzer (Randox).The markers were analysed against high risk karyotype, remission achievement after induction, age over 65 years at diagnosis, highly elevated lactate dehydrogenase (LDH) – exceeding 5 times upper limit of normal values in our laboratory, PFS and OS.Results:Higher age and high risk genetics were related to disease progression which was not significant after Bonferroni‐Holm correction (P = 0.083 and 0.132 respectively). In low and intermediate risk group, higher age was associated with shorter PFS (P < 0.001). Disease progression was strongly associated with death. Those who were progression‐free for at least two years had much lower risk of relapse, (P < 0.001) for the whole cohort. Allogeneic stem cell transplantation was associated with survival benefit (uncorrected P < 0.001). From evaluated serum factors, TNFR‐1 and sIL‐6R had relationship with PFS, but lost statistical significance after Bonferroni‐Holm correction.OS was influenced by age (P = 0.033), high‐risk cytogenetics (P = 0.001), hematologic relapse (P < 0.001) and prolonged relapse‐free period (P < 0.001). In the whole cohort, analysis CR after induction therapy was not associated with survival benefit, which was also true for low and intermediate risk subgroup. In high‐risk subgroup, the CR after induction was associated with survival benefit (P < 0.001). Highly elevated LDH was a negative prognostic factor (P = 0.012) in low and intermediate risk subgroup. In the whole cohort, the levels of sIL‐2Rα (corrected P = 0.001, Figure 1) and TNFR‐1 (corrected P = 0.053) were candidate factors for improvement of OS prediction.Summary/Conclusion:Understanding the cancer microenvironment may help in development of new treatment approaches. Our results show that sIL‐2Rα and TNFR‐1 may be associated with treatment outcome and thus should be further investigated as possible therapeutic targets.imageThe work was supported by a long‐term organisation development plan 1011 (FMHS).

Increased Kynurenine Plasma Concentrations and Kynurenine-Tryptophan Ratio in Mild-To-Moderate Chronic Obstructive Pulmonary Disease Patients

Since an increase in kynurenine (Kyn) plasma concentrations has been proposed as marker of immune system activation, we studied the associations between the Kyn levels and presence and severity of chronic obstructive pulmonary disease (COPD). Plasma Kyn, tryptophan (Trp) and Kyn/Trp ratio were measured in 43 COPD patients with clinically defined mild (n=29) or moderate (n=14) disease and 43 age- and sex-matched healthy controls. When compared with controls, COPD patients had significantly higher plasma Kyn concentrations and Kyn/Trp ratios. In multiple logistic regression analysis, after adjusting for clinical and demographic confounders, the Kyn/Trp ratio was independently associated with COPD severity. Kyn and Kyn/Trp ratio might represent a new, sensitive, biomarker of systemic inflammation in COPD patients.

Serum Levels of Cytokines and Cytokine Receptors Are Associated with Outcome in Newly Diagnosed AML Patients

Background: Cytokines, cytokine receptors and adhesion molecules have been studied as markers of immune system activation in various diseases including AML/MDS. These factors form a dynamic network which affects AML cell susceptibility to chemotherapy. To provide more evidence of these interactions, we analyzed the serum levels of a broad panel of cytokines, cytokine receptors, matrix metalloproteinase-9 and soluble adhesion molecules.Aims: The aim of our study was to evaluate hypothesis that baseline serum levels of these factors are associated with relapse and overall survival (OS).Methods: A total of 65 AML patients (27 males, 38 females, mean age 53.7 ± 12.8, median 57.1 years) newly diagnosed in the period 2011 - 2014 were analyzed. All patients were Caucasian. Of these, 14 had better risk, 10 intermediate-1 risk, 13 intermediate-2 risk and 28 high risk AML according to ELN risk stratification (Döhner et al., Blood 2010). The NPM-1 was mutated in 16 cases, the FLT3-ITD was present in 15 cases. The secondary disease was present in 26 cases, 3 patients were BCR/ABL positive. Five patients with APL were treated with the PETHEMA regimen combining Idarubicin and All-trans retinoic acid (ATRA). All other patients were induced with "3+7" induction chemotherapy consisting of Cytarabin 100mg/m2 per day for 7 consecutive days and Daunorubicin 90mg/m2 for the first 3 days of therapy in younger patients. In patients aged ≥ 65 years, Daunorubicin 45-60mg/m2 was administered, depending on patient WHO performance status.We evaluated baseline circulating levels of the following factors: interleukins (IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12p70, IL-13, IL-15), epidermal growth factor (EGF), granulocyte-macrophage colony stimulating factor (GM-CSF), interferon-gamma (IFN-γ), monocyte chemotactic protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α), matrix-metalloproteinase-9 (MMP-9), tumor necrosis factor-alpha (TNF-α), vascular endothelial growth factor (VEGF), soluble IL-2 receptor-α (sIL-2Rα) and soluble receptors for IL-6 (sIL-6R) and TNF-α type I and II (TNFR-1,2), E-selectin (E-SEL), P-selectin (P-SEL), intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1). All biomarkers were measured by biochip array technology on Evidence Investigator analyzer (Randox). Statistical analysis was performed in R 3.1.2. Probability values (P) < 0.05 were considered statistically significant.Results: All evaluated analytes were independent of age, disease origin and ELN risk stratification. Relapse occurred in 34 cases with median of 516 days. Median follow-up was 627 days. Primary AML, normal karyotype and achieving of complete remission (CR) after induction therapy were associated with better relapse free survival (RFS), whereas higher TNFR-1, VEGF, IL-3, TNFR-2, NPM-1 and BCR/ABL were associated with inferior RFS. OS correlated with levels of IFN-γ, GM-CSF and TNFR-1. In multivariate analysis, primary AML, higher IL-10 and IL-15 were associated longer OS. Higher age, higher ELN risk and higher levels of TNFR-1, IL-3, IL-6, IL-8 and TNFR-2 were associated with inferior OS.Conclusions: Cytokines are an important part of cancer environment. In AML, some cytokines or their receptors may be informative for individual prognosis. The literature suggests association of IL-6 and IL-10 levels and OS (Correa et al., Cytokine 2013). Our data are in agreement with this finding, but revealed more cytokines and their receptors, especially TNFR-1, which were independently associated with relapse and OS. To confirm these findings, analysis in a larger cohort with longer follow-up should be performed.Acknowledgment:The work was supported by a specific research project "Analysis of defined prognostic factors in acute myeloid leukemia" (FMHS Hradec Kralove), by a long-term organization development plan 1011 (FMHS Hradec Kralove) and by MH CZ - DRO (UHHK, 00179906). DisclosuresNo relevant conflicts of interest to declare.

Cholestatic Liver Injury as a Feature of Hemophagocytic Lymphohistiocytosis

We present the case of a 29-year-old male with Crohn's disease on 5-ASA/6MP therapy who was admitted for 11 days of worsening fevers, cough, chest pain and malaise. He had no recent travel or sick contacts and had recently tested negative for HIV. On admission, he was taking levofloxacin per his PCP for suspected pneumonia, but noted that his symptoms had not improved at all. His admission labs revealed pancytopenia and he continued to spike fevers up to 40°C. His AST/ALT/ALP/Total bilirubin were 125/58/131/1.2, and an abdominal ultrasound on admission showed a distended gallbladder wall and pericholecystic fluid with no evidence of calculi, suggestive of acalculous cholecystitis. He was started on vancomycin, aztreonam and metronidazole while awaiting blood culture results and further workup. His blood cultures came back negative, but his LFTs progressively increased to 213/67/251/5.7 to reveal a cholestatic picture. A subsequent HIDA was negative and a CT of the abdomen and pelvis showed only hepatosplenomegaly. His serologies confirmed immunity to Hepatitis B virus and non-exposure to Hepatitis A and C viruses, and he had no evidence of cirrhosis on history or physical exam. He was found to have CMV/ EBV viremia, which normally causes hepatocellular injury, so the reason for his cholestatic injury still remained mysterious at this point. A bone marrow biopsy performed to elucidate the cause of his pancytopenia revealed marked hemophagocytosis. Further testing revealed a significantly elevated ferritin level at 11700 ng/ml (normal < 336 ng/ml). His sCD25, a marker of immune system activation and inflammation, was grossly elevated as well. The patient was eventually diagnosed with hemophagocytic lymphohistiocytosis (HLH) on the basis of his fevers ≥38.5°C, splenomegaly, peripheral blood cytopenias, hyperbilirubinemia, hemophagocytosis in the bone marrow, ferritin > 3000 ng/ml, and elevated sCD25. He was started on treatment with etoposide and dexamethasone, with eventual resolution of his cholestatic liver injury. He is now awaiting results of a genetic panel to screen for HLH-causing mutations. HLH is a rare syndrome that occurs due to immune system dysregulation, often triggered by infection in susceptible individuals. In HLH, NK cells and cytotoxic lymphocytes fail to downregulate activated macrophages, resulting in a cytokine storm. High levels of IFN-γ and other cytokines cause severe tissue damage and organ failure, necessitating prompt management.

Association of neopterin as a marker of immune system activation and juvenile rheumatoid arthritis activity

ObjectiveTo evaluate neopterin plasma concentrations in patients with active juvenile idiopathic arthritis (JIA) and correlate them with disease activity. MethodsSixty patients diagnosed as active JIA, as well as another 60 apparently healthy age‐ and gender‐matched children as controls, were recruited from the Pediatrics Allergy and Immunology Clinic, Ain Shams University. Disease activity was assessed by the Juvenile Arthritis Disease Activity Score 27 (JADAS‐27). Laboratory investigations were performed for all patients, including determination of hemoglobin concentration (Hgb), erythrocyte sedimentation rate (ESR), and C‐reactive protein. Serum concentrations of tumor necrosis factor‐alpha (TNF‐α), interleukin‐6 (IL‐6), monocyte chemoattractant protein‐1 (MCP‐1), and neopterin were measured. ResultsSignificant differences were found between JIA patients and controls with regard to the mean levels of Hgb, ESR, TNF‐α, IL‐6, and MCP‐1 (p<0.05). A statistically significant higher mean level serum neopterin concentration (p<0.05) was found in JIA patients (20.43±8.73 nmol/L) than in controls (6.88±2.87 nmol/L) (p<0.05). Positive significant correlations were detected between serum neopterin and ESR, TNF‐α, IL‐6, MCP‐1, and JADAS‐27 (p<0.05). No correlation was found between serum neopterin and CRP (p>0.05). Multiple linear regression analysis showed that JADAS‐ 27 and ESR were the main variables associated with serum neopterin in JIA patients (p<0.05). ConclusionThe elevation of plasma neopterin concentrations in early JIA patients may indicate stimulation of immune response. Serum neopterin can be used as a sensitive marker for assaying background inflammation and disease activity score in JIA patients.

Association of neopterin as a marker of immune system activation and juvenile rheumatoid arthritis activity

ObjectiveTo evaluate neopterin plasma concentrations in patients with active juvenile idiopathic arthritis (JIA) and correlate them with disease activity. MethodsSixty patients diagnosed as active JIA, as well as another 60 apparently healthy age- and gender-matched children as controls, were recruited from the Pediatrics Allergy and Immunology Clinic, Ain Shams University. Disease activity was assessed by the Juvenile Arthritis Disease Activity Score 27 (JADAS-27). Laboratory investigations were performed for all patients, including determination of hemoglobin concentration (Hgb), erythrocyte sedimentation rate (ESR), and C-reactive protein. Serum concentrations of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), monocyte chemoattractant protein-1 (MCP-1), and neopterin were measured. ResultsSignificant differences were found between JIA patients and controls with regard to the mean levels of Hgb, ESR, TNF-α, IL-6, and MCP-1 (p<0.05). A statistically significant higher mean level serum neopterin concentration (p<0.05) was found in JIA patients (20.43±8.73 nmol/L) than in controls (6.88±2.87 nmol/L) (p<0.05). Positive significant correlations were detected between serum neopterin and ESR, TNF-α, IL-6, MCP-1, and JADAS-27 (p<0.05). No correlation was found between serum neopterin and CRP (p>0.05). Multiple linear regression analysis showed that JADAS- 27 and ESR were the main variables associated with serum neopterin in JIA patients (p<0.05). ConclusionThe elevation of plasma neopterin concentrations in early JIA patients may indicate stimulation of immune response. Serum neopterin can be used as a sensitive marker for assaying background inflammation and disease activity score in JIA patients.

Impact of cholesterol lowering treatment on plasma kynurenine and tryptophan concentrations in chronic kidney disease: Relationship with oxidative stress improvement

Background and aimTryptophan (Trp) degradation via indoleamine (2,3)-dioxygenase (IDO), with consequent increased in kynurenine (Kyn) concentrations, has been proposed as marker of immune system activation. Oxidative stress (OS) might contribute to the pro-inflammatory state in chronic kidney disease (CKD) through the activation of NF-kB, with consequent activation and recruitment of immune cells. Methods and resultsSerum concentrations of Trp and Kyn, oxidative stress indices malondialdehyde (MDA) and allantoin/uric acid (All/UA) ratio and anti-oxidant amino acid taurine were measured in 30 CKD patients randomized to 40 mg/day simvastatin (group 1), ezetimibe/simvastatin 10/20 mg/day (group 2) or ezetimibe/simvastatin 10/40 mg/day (group 3) and treated for 12 months. Baseline Kyn and Kyn/Trp ratio were higher in CKD patients vs. healthy controls (1.67 ± 0.62 μmol/L vs 1.25 ± 0.40 μmol/L, p < 0.01 and 0.036 ± 0.016 vs 0.023 ± 0.010, p < 0.001 respectively). Both Kyn and Kyn/Trp ratio significantly decreased after cholesterol lowering treatment, to values comparable with healthy controls after one year treatment (1.67 ± 0.62 μmol/L vs 1.31 ± 0.51 μmol/L, p < 0.0001 and 0.036 ± 0.016 vs 0.028 ± 0.012 p < 0.0001, respectively). This was paralleled by a significant decrease in MDA (218 ± 143 nmol/L vs 176 ± 123 nmol/L, p < 0.01) and All/UA ratio (1.47 ± 0.72 vs 1.19 ± 0.51, p < 0.01) in CKD patients. ConclusionsAmelioration of both oxidative and inflammation status after cholesterol lowering treatment in CKD might be mediated by restoration of antioxidant taurine concentrations during therapy (from 51.1 ± 13.3 μmol/L at baseline to 63.1 ± 16.4 μmol/L, p < 0.001 by ANOVA), suggesting that improvement of both oxidative and inflammation status in CKD patients could be explained, at least partly, by the cholesterol lowering effects.

The role of neopterin in patients with primary and metastatic breast cancer: correlation with clinicopathological data

Breast cancer is the commonest malignancy of women in many parts of the world and comprises 18% of all female cancers. There is one million new cases in the world each year. Neopterin which is a pteridine produced by activation of monocytes by IFN-γ may be used as a marker of immune system activation. The biological function of neopterin is still largely unknown and certain patho-physiologic roles have only recently been elucidated. Therefore, the aim of this work was to investigate the role of neopterin in females with primary and metastatic breast cancer. The obtained results were correlated with different clinicopathological data of those patients. The study population included females with primary and metastatic breast cancer as well as healthy age matched females were taken as a control group. Peripheral blood mononuclear cells were separated from each subject under study by Ficoll-Hypaque density gradient centrifugation technique. The isolated cells were cultured with and without PHA and neopterin levels were measured in serum and culture supernatants by ELISA technique. The results showed that neopterin levels were lowest in serum, followed by culture supernatants without PHA and highest in culture supernatants with PHA in each studied group. The mean neopterin values in serum and culture supernatants with PHA of metastatic breast cancer patients were significantly higher than in the primary breast cancer patients and the control group. Our results also indicated that neopterin levels positively correlated with the tumor grade, tumor stage and the presence of metastasis. From all mentioned data it is clear that, measurement of neopterin either in serum or in culture supernatants with or without PHA can give an indication about immunological status of breast cancer patients' response to therapy and early detection of metastasis. It gives an important and valuable prognostic information before surgical intervention is performed.

Serum Profile Of Multiple Cytokines and Adhesion Molecules In Newly Diagnosed AML Patients Differs In Those Who Did Not Reach Complete Remission After Anthracycline/Cytarabine Induction Therapy

BackgroundCytokines and adhesion molecules have been studied as markers of immune system activation in various diseases including AML/MDS. Cytokines are soluble molecules taking part in intercellular communication with a specific role in cell proliferation and differentiation control. Further knowledge gained from multiple cytokine and adhesion molecule analysis should allow better diagnosis and disease management. AimsThe aim of our study was to evaluate baseline serum levels of multiple cytokines and adhesion molecules in Caucasian population AML patients and compare it with standard prognostic indicators in patients with AML. MethodsA total of 28 consecutive newly diagnosed AML patients (11 males, 17 females, age 21 to 71 years, mean 52.3 ± 12.1, median 55.4 years, 8 with better risk, 13 with intermediate risk, 7 with high risk according to cytogenetics and molecular genetics) were administered 3+7 induction regimen with escalated dose of daunorubicin 90mg/m2. We evaluated serum levels of the following 22 cytokines and adhesion molecules: interleukins (IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-4, IL-6, IL-7, IL-8, IL-10, IL-12, IL-13, IL-23), vascular endothelial growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), interferon-gamma (IFN-gamma), epidermal growth factor (EGF), monocyte chemotactic protein-1 (MCP-1), E-selectin, L-selectin, P-selectin, intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1). All biomarkers were measured by biochip array technology on Evidence Investigator analyzer (Randox) at the diagnosis of AML and compared with standard prognostic indicators in AML. Probability values (p) < 0.05 were considered statistically significant. ResultsAs there was no significant difference between younger and elderly patients, we evaluated the cohort as a whole. Comparing serum cytokine and adhesion molecule levels in patients who did not achieve CR after induction therapy (n=6, 21.5%) to those who achieved CR (n=22, 78,5%) a statistically significant increase in serum IL-2 (5.42 ± 5.42 ng/L vs. 2.02 ± 1.84 ng/L; p < 0,05), IL-7 (11.71 ± 10.77 ng/L vs. 3.50 ± 1.51 ng/L; p < 0.05), IL-8 (241.9 ± 225.3 ng/L vs. 34.31 ± 25.47 ng/L; p < 0.005), IL-10 (7.74 ± 6.83 ng/L vs. 2.83± 2.68 ng/L; p < 0.05) was found. Both groups differed from healthy individuals in serum levels of IL-4, IL-6, IL-13, VCAM-1, ICAM-1, E-selectin and L-selectin. We found moderate inverse correlation between overall survival and levels of VCAM-1 and E-selectin. There was no significant difference in cytokines and adhesion molecules levels among established prognostic subgroups in AML. ConclusionsThe results in general are in agreement with outcome of our pilot trial. Compared with the other studies of cytokine levels in newly diagnosed AML, our data indicate different conclusions. IL-2, IL-4 and IL-10 were more likely to be higher in those who attained remission (Kornblau et al., Blood 2010). We did not find association between IL-6 and IL-10 levels and survival in our studied group (Correa et al, Cytokine 2013). On the other hand we documented inverse correlation between levels of some adhesion molecules and overall survival. Whether these alterations depend on the studied population is not known definitely. To assess their predictive value for patient outcome, further studies in a larger number of patients is necessary. AcknowledgmentThe work was supported by Specific research project “Analysis of defined prognostic factors in acute myeloid leukemia” (FMHS) and by a long-term organisation development plan 1011 (FMHS). Disclosures:No relevant conflicts of interest to declare.

Correlation between DAS-28 and neopterin as a biochemical marker of immune system activation in early rheumatoid arthritis

Objective: The objective of this study was to evaluate neopterin plasma concentrations in patients with early Rheumatoid Arthritis (RA) and correlate them with disease activity. Methods: This is a 28-month prospective study carried out on 65 individuals. There were 27 patients with early RA and 38 healthy volunteers as control group. Enzyme immunoassay was used to measure concentrations of neopterin and anti-cyclic citrullinated peptide (CCP) antibodies. Rheumatoid factor (RF) and C-reactive protein (CRP) were measured turbidimetrically, and antinuclear antibodies (ANA) were detected by immunofluorescence. Patients with early RA disease activity were divided into 4 groups according to DAS-28 criteria. Neopterin concentrations in RA patients were compared to conventional RA diagnostic serological markers. Results: Healthy volunteers had a mean neopterin concentration of 5.63 ± 0.38 [1.36–9.93] nmol. L− 1. A statistically significant elevation of neopterin mean concentration was found on early RA patients: mean value of 8.92 ± 0.93 [3.94–28.3] nmol. L− 1 (p < 0.001). Pearson product moment correlation suggests a correlation between neopterin concentrations and DAS-28 (r = 0.208, p = 0.065). The analysis of the mean values grouped according with the DAS-28 criteria showed a correspondence between these means, with a Pearson correlation coefficient r = 0.979, p = 0.021. CRP concentrations also showed a similar trend. Anti-CCP antibodies and RF revealed a positive relationship with RA activity. Such a correlation was not found with ANA results. Conclusions: The elevation of plasma neopterin concentrations in early RA patients may indicate stimulation of immune response. Good correlation between neopterin concentrations and DAS-28 may facilitate assessing disease activity.

Elevated Serum Soluble CD30 Precedes the Development of AIDS-Associated Non-Hodgkin’s B Cell Lymphoma

CD30, first described as the Ki antigen on malignant B cells in Hodgkin’s lymphoma, is also expressed on normal activated B and T cells. It can be cleaved from the cell surface and detected in normal serum as soluble CD30 (sCD30), where it can be an indicator of levels of immune activation. In a cross-sectional study utilizing archived sera at a time point close to but preceding a diagnosis of acquired immunodeficiency syndrome (AIDS)-associated non-Hodgkin’s B cell lymphoma, AIDS lymphoma subjects (n = 49) showed elevated mean levels of sCD30 compared to controls with AIDS but no malignancy (n = 44, p < 0.01), HIV-infected but relatively healthy (n = 47, p < 0.001), or HIV-seronegative controls (n = 44, p < 0.001). Serum sCD30 was significantly correlated to serum levels of the B cell cytokines interleukin-6 (IL-6), IL-10, and sCD23, but only among lymphoma subjects (p ≤ 0.05). Correlations between sCD30 and other markers of immune system activation were seen among all HIV-infected subjects (sCD27, sCD44, CXCL13, p < 0.05). These observations suggest that sCD30, especially in combination with other immune system molecules, could be an important biomarker for an immune system environment conducive to B cell hyperactivation and the development of AIDS-associated B cell lymphoma.

A C-reactive protein promoter polymorphism is associated with type 2 diabetes mellitus in Pima Indians

Linkage analysis has identified a susceptibility locus for type 2 diabetes mellitus (T2DM) on chromosome 1q21–q23 in several populations. Results from recent prospective studies indicate that increased levels of C-reactive protein (CRP), a marker of immune system activation, are predictive of diabetes, independent of adiposity. Because CRP is located on 1q21, we considered it a potential positional candidate gene for T2DM. We therefore evaluated CRP and the nearby serum amyloid P-component, APCS, which is structurally similar to CRP, as candidate diabetes susceptibility genes. Approximately 10.9 kb of the CRP-APCS locus was screened for polymorphisms using denaturing high performance liquid chromatography and direct sequencing. We identified 27 informative polymorphisms, including 26 single nucleotide polymorphisms (SNPs) and 1 insertion/deletion, which were divided into 7 linkage disequilibrium clusters. We genotyped representative SNPs in ∼1300 Pima samples and found a single variant in the CRP promoter (SNP 133552) that was associated with T2DM ( P=0.014), as well as a common haplotype (CGCG) that was associated with both T2DM ( P=0.029) and corrected insulin response, a surrogate measure of insulin secretion in non-diabetic subjects ( P=0.050). Linkage analyses that adjusted for the effect of these polymorphisms indicated that they do not in themselves account for the observed linkage with T2DM on chromosome 1q. However, these findings suggest that variation within the CRP locus may play a role in diabetes susceptibility in Pima Indians.

Serum and salivary neopterin and interferon‐γ in primary Sjögren's syndromeCorrelation with clinical, laboratory and histopathologic features

Objective: To investigate serum and salivary neopterin and interferon‐γ as possible markers of immune system activation in primary Sjögren's Syndrome (pSS).Methods: Serum and salivary neopterin and interferon‐γ concentrations were determined in 30 untreated patients with pSS and matched with several other clinical and laboratory parameters.Results: The mean concentration of neopterin was significantly higher in pSS patients (8.12±3.36 nmol/L in serum and 9.50±7.61 nmol/L in saliva) than in normal controls (p<0.05). Significant correlations were found between serum neopterin and β2‐microglobulin, serum IgG as well as lip biopsy score. Salivary neopterin concentration was inversely related to Shirmer‐I test, tear break‐up time and stimulated salivary flow rate. Serum and salivary levels of interferon‐γ were normal and no correlation with the other parameters was found.Conclusion: In pSS patients serum neopterin may represent a useful marker of cell‐mediated immunity. On the other hand, salivary neopterin seems to reflect the glandular damage.

Ocular features associated with anticardiolipin antibodies: A descriptive study

PURPOSE: To evaluate ocular features in patients presenting with inflammation in the presence of anticardiolipin antibodies. METHODS: A descriptive study of 13 patients presenting with idiopathic ocular inflammation involving anterior and posterior segment was performed. Patients were followed for a mean follow-up of 22 months (range, 1 to 125). A comprehensive report of ocular involvement, including visual symptoms, visual acuity, clinical characteristics, funduscopic and fluorangiographic features, was reported. Systemic associated symptoms were analyzed. Laboratory investigations included anticardiolipin antibody titers and isotypes, presence of other autoantibodies, and markers of immune system activation. RESULTS: The most common ocular symptom at presentation was blurred vision (eight patients) followed by redness and pain(three patients) and visual loss(two patients). Anterior segment abnormalities, including iritis (eight patients) scleritis (two patients) and filamentary keratitis (one patient), were present in 76% of patients, whereas the most represented feature of posterior involvement was retinal vasculitis (60%) followed by vitritis (38%), retinal detachment (15%), posterior scleritis (7%), and central retinal artery occlusion (7%). All patients had abnormal titers of anticardiolipin antibodies, predominantly IgG isotype; six had markers of immune system activation. CONCLUSIONS: Although posterior pole disease is more commonly associated with anticardiolipin antibodies, the anterior segment can also be involved with a wide spectrum of features. Scleritis has never been previously described as associated with anticardiolipin antibodies. Systemic symptoms are frequently present in association with ocular disease.

A dual pathway model of daily stressor effects on rheumatoid arthritis.

This study evaluated the initial promise of a dual-pathway conceptual model linking daily event stressors to rheumatoid arthritis (RA) disease activity through changes in immune system activation and mood. Fifty individuals, who were studied on five occasions two weeks apart, reported daily event stressors on the Daily Life Experience Checklist, daily mood on an abbreviated version of the Profile of Mood States-B, and daily joint pain on the Rapid Assessment of Disease Activity in Rheumatology. Serial clinical examinations comprised ratings of joint tenderness and swelling, and blood drawn during exams was analyzed for sedimentation rate (an indicator of systemic inflammation) and soluble interleukin-2 receptors (a marker of immune system activation known to correlate with RA disease activity). Across-person analyses failed to establish links from daily event stressors to either disease activity or composites of joint pain and joint inflammation when associations were adjusted for the effect of neuroticism on self-report measures. Pooled within-person analyses, however, were generally consistent with the relations predicted by the dual-pathway model. Increases in daily event stressors during the week preceding each clinical exam were associated with increased joint pain (regardless of changes in mood). At the same time, increased daily stressors were indirectly associated with decreased joint inflammation through reduction in levels of soluble interleukin-2 receptors. The dual-pathway model, which may be limited to short-term psychological and psychoimmunologic processes, underscores the importance of distinguishing potentially opposing effects of stress on pain versus inflammation in individuals with rheumatoid arthritis.

Temporal variability in immunological parameters: Peripheral blood mononuclear cell subsets, serum immunoglobulins, and soluble markers of immune system activation

T-cell subsets and soluble factors of immune system activation are increasingly used as biologic markers of disease and predictors of disease progression. For example, changes in CD4 cells and CD4:CD8 ratio, sIL-2R, B2M, neopterin, and IgA have been used in predicting AIDS onset and progression. We examined the temporal variability of T-cell subsets, monocytes, natural killer cells, B cells, immunoglobulins, soluble interleukin-2 receptor (sIL-2R), neopterin, and beta-2 microglobulin (B2M) among 135 adults tested at two time points approximately 3 months apart. The purpose of the study was two-fold: (1) to assess the stability of these measures at two points in time, and (2) to investigate which parameters tend to track together over time, i.e., show significant longitudinal correlation. Mean population values for these immunologic parameters remained remarkably stable over the 3-month period. However, individual subjects exhibited significant temporal variability for many parameters. Unlike observations in patients with AIDS, changes in immunoglobulins and other soluble factors were not significantly correlated with changes in cellular subsets over the same period. However, change in B2M was correlated with change in neopterin (r = .35, p < or = .0001), and change in IgA was correlated with changes in IgG and IgM (r = .44, r = .54, P < or = .001 for both). Characterizing this temporal variability in a healthy population provides important information for researchers applying these tests in clinical and epidemiological studies.