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PDF file - 109K, CD4+CD8dim or CD4+NKG2D+ T cells can be found in other malignancies.

PDF file - 84K, The combination therapy induced a profound lymphopenia.

PDF file - 38K, Synergistic effects of TCR or IL-15 stimulation with NKG2D engagement for Th1 polarization in CD4+NKG2D+T cells.

PDF file - 94K, 2D Clustering of LogRatios (D21/D0) of gene expression post- (D21) and pre-therapy (D0) with sorafenib/temozolomide.

PDF file - 1795K, T-cell infiltration and MICA/B expression in metastatic melanoma.

Beyond cancer-cell intrinsic factors, the immune status of the host has a prognostic impact on patients with cancer and influences the effects of conventional chemotherapies. Metastatic melanoma is intrinsically immunogenic, thereby facilitating the search for immune biomarkers of clinical responses to cytotoxic agents. Here, we show that a multi-tyrosine kinase inhibitor, sorafenib, upregulates interleukin (IL)-15Rα in vitro and in vivo in patients with melanoma, and in conjunction with natural killer (NK) group 2D (NKG2D) ligands, contributes to the Th1 polarization and accumulation of peripheral CD4+NKG2D+ T cells. Hence, the increase of blood CD4+NKG2D+ T cells after two cycles of sorafenib (combined with temozolomide) was associated with prolonged survival in a prospective phase I/II trial enrolling 63 patients with metastatic melanoma who did not receive vemurafenib nor immune checkpoint–blocking antibodies. In contrast, in metastatic melanoma patients treated with classical treatment modalities, this CD4+NKG2D+ subset failed to correlate with prognosis. These findings indicate that sorafenib may be used as an “adjuvant” molecule capable of inducing or restoring IL-15Rα/IL-15 in tumors expressing MHC class I–related chain A/B (MICA/B) and on circulating monocytes of responding patients, hereby contributing to the bioactivity of NKG2D+ Th1 cells. Cancer Res; 74(1); 68–80. ©2013 AACR.

PDF file - 25K, Schedule of the sorafenib and temozolomide combination regimen.

PDF file - 119K

BACKGROUND: Nurses play a central role in the management of atrial fibrillation (AF) patients. An unresolved question is whether a nurse-led clinic would improve clinical outcomes. Herein, we investigated the impact of a nurse-led clinic on anticoagulation therapy and clinical outcomes in a cohort of naïve AF patients.METHODS: Prospective study including AF patients starting vitamin K antagonists (VKAs) into a nurse-led AF clinic. These patients were followed in this specific AF clinic. Additionally, AF patients already taking VKAs for 6 months followed according to the routine clinical practice were included as comparison group. The quality of anticoagulation was assessed at 6 months. Efficacy and safety endpoints were recorded during follow-up.RESULTS: We included 223 patients (Nurse-led clinic: 107; Usual care: 116). The mean time in therapeutic range and the proportion of INRs within the therapeutic range were similar in both groups. During 2.06 (IQR 1.01-2.94) years of follow-up, 64 (28.7%) patients changed to direct-acting oral anticoagulants. The proportion of switchers was higher in the nurse-led clinic (37.4%) than in the usual care group (20.7%) (P = .006) and these patients spent less time to switch (2.0 [IQR 0.7-2.9] vs 6.0 [IQR 3.7-11.2] years; P < .001). Importantly, the annual rate of ischaemic stroke/TIA was significantly lower in the nurse-led clinic (0.47%/year vs 3.88%/year, P = .016), without differences in safety endpoints.CONCLUSION: A nurse-led AF clinic may offer a "patient-centered" review and holistic follow-up, and it would be associated with a reduction of ischaemic stroke/TIA, without increasing bleeding complications. Further studies should confirm these results.

Background Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations estimate glomerular filtration rate (GFR) more accurately than the Modification of Diet in Renal Disease (MDRD) equation. Hypothesis New CKD-EPI equations improve risk stratification in patients with non-ST-segment elevation acute coronary syndrome (NSTE-ACS) and provide complementary information to the Global Registry of Acute Coronary Events (GRACE) risk score. Methods We studied 350 subjects (mean age, 68 ± 12 years; 70% male) with NSTE-ACS. Estimated GFR was calculated using the MDRD and new CKD-EPI equations based on serum creatinine (SCr) and/or cystatin C (CysC) concentrations obtained within 48 hours of hospital admission. The primary endpoint was all-cause death during follow-up. Results Over the study period (median, 648 days [interquartile range, 236–1042 days]), 31 patients died (0.05% events per person-year). Decedents had poorer renal-function parameters (P < 0.001). Both CysC-based CKD-EPI equations had the highest areas under the receiver operating characteristic curve for the prediction of all-cause mortality. After multivariate adjustment, only CysC-based CKD-EPI equations were independent predictors of all-cause mortality (CKD-EPISCr-CysC, per mL/min/1.73 m2: hazard ratio: 0.975, 95% confidence interval: 0.956-0.994, P = 0.009; CKD-EPICysC, per mL/min/1.73 m2: hazard ratio: 0.976, 95% confidence interval: 0.959-0.993, P = 0.005). Reclassification analyses showed that only CysC-based CKD-EPI equations improved predictive accuracy of the GRACE risk score. Conclusions In patients with NSTE-ACS, CysC-based CKD-EPI equations improved clinical risk stratification for mortality and added complementary prognostic information to the GRACE risk score.

Background A simple method to assess renal function is the estimated glomerular filtration rate, and it shows prognostic implications. However, it remains unknown which equation should be used in patients with acute coronary syndrome. We compared the ability and correlation of the Cockcroft‐Gault, Modification of Diet in Renal Disease‐4 (MDRD‐4), and Chronic Kidney Disease Epidemiology Collaboration (CKD‐EPI) equations and their predictive performance for major adverse cardiovascular events, all‐cause mortality, and major bleeding in a cohort of patients with acute coronary syndrome. Methods and Results Multicenter prospective registry involving 1699 consecutive patients with acute coronary syndrome from 3 tertiary institutions. At entry, renal function was assessed using the Cockcroft‐Gault, MDRD ‐4, and CKD ‐ EPI ‐creatinine equations. During 12 months of follow‐up, we recorded all major adverse cardiovascular events (composite of cardiovascular death, nonfatal myocardial infarction, and nonfatal ischemic stroke), bleeding events (Bleeding Academic Research Consortium classification), and all‐cause mortality. Receiver operating characteristic curve comparisons demonstrated that Cockcroft‐Gault equation had higher predictive ability compared with MDRD ‐4 equation for major adverse cardiovascular events (0.651 versus 0.616; P =0.023), major bleeding (0.600 versus 0.551; P =0.005), and all‐cause mortality (0.754 versus 0.717; P =0.033), as well as higher predictive ability compared with CKD ‐ EPI equation for major bleeding (0.600 versus 0.564; P =0.018). Integrated discrimination improvement and net reclassification improvement analyses showed superior discrimination and reclassification of Cockcroft‐Gault equation. Decision curve analyses graphically demonstrated higher net benefit and clinical usefulness of the Cockcroft‐Gault equation in comparison with MDRD ‐4 and CKD ‐EPI equations. Conclusions In patients with acute coronary syndrome, the Cockcroft‐Gault equation presented superior predictive ability for major adverse cardiovascular events, major bleeding, and all‐cause mortality compared with MDRD ‐4 equation, and superior predictive ability for major bleeding compared with CKD ‐EPI equation. The Cockcroft‐Gault equation also showed higher net benefit and clinical usefulness.

Atrial fibrillation (AF) is the most common sustained chronic cardiac arrhythmia in clinical practice, which increases the risk of stroke and thromboembolism and is an independent predictor of mortality. The underlying mechanisms involved in the development of AF have yet to be fully elucidated. However, once initiated, AF tends to self-perpetuate, owing to structural and electrical remodeling in the atria. MicroRNAs (miRNAs) represent a sizable sub-group of small non-coding RNAs, which degrades or inhibits the translation of their target mRNAs, thus regulating gene expression and playing an important role in a wide range of biologic processes. Clinically, there is increasing evidence of the potential diagnostic role of miRNAs as biomarkers, representing a novel therapeutic target in AF. The aim of this review is to provide an exhaustive overview of the role of miRNAs in AF and to discuss the diagnostic and therapeutic potential of miRNAs in this arrhythmia.

Resumen Introduccion: Lafibrilacionauricular(FA),conunaincidenciaaproximadadel30%,eslaarritmiamas frecuente tras cirugia cardiaca. Se han asociado a la FA factores como la inflamacion, lapresencia de fibrosis cardiaca, el estres y la apoptosis de cardiomiocitos. Objetivos: Consideramos que el remodelado auricular es un proceso preexistente en los pacien-tes con FA posquirurgica. Analizamos los factores relacionados con la incidencia de FA en elpostoperatorio de cirugia cardiaca. Metodos: Incluimos a pacientes consecutivos, estables hemodinamicamente y en ritmo sinusal,sometidos a cirugia cardiaca programada con circulacion extracorporea. Se valora la caida enFA posquirurgica. Resultados: Se incluyeron un total de 100 pacientes sometidos a cirugia de revascularizacioncoronaria (59) o sustitucion valvular aortica (41) por estenosis aortica grave. La FA postope-ratoria se produjo en 29 pacientes con predominio de la cirugia valvular respecto a la cirugiacoronaria. Los factores predictivos de la aparicion de FA postoperatoria en el analisis multi-variable fueron el sexo masculino, la ausencia de terapia cronica con betabloqueadores, laperfusion de fibrinogeno intraoperatorio, valores bajos de colesterol HDL y valores elevados detroponina T ultrasensible en el preoperatorio.

Background Hypertrophic cardiomyopathy (HCM) is characterized by inappropriate hypertrophy, myocyte disarray and increased interstitial fibrosis. The tumour necrosis factor-like weak inducer of apoptosis (TWEAK) is a cell surface cytokine with biological activities including stimulation of cell growth, induction of inflammatory cytokines and stimulation of apoptosis. There are controversial data about the potential role of TWEAK in different cardiovascular pathologies. NT-proBNP is an established biomarker of myocardial wall stress, associated with poor functional class in HCM. We hypothesized that effort capacity in patients with HCM could be related to serum levels of these biomarkers. Materials and methods We included 40 haemodynamic stable HCM patients and 53 healthy controls with similar sex and age. We studied exercise capacity by maximal oxygen consumption in a limited treadmill exercise test. TWEAK and NT-proBNP were assayed by ELISA method and automated Elecsys® platform, respectively. We obtained 46 samples of myocardial tissues by septal myectomy in patients with HCM and evaluated myocardial fibrosis, immunoreaction with TWEAK antibody and apoptosis with TUNEL assay. Results We found raised TWEAK and NT-proBNP serum levels in patients when compared with control levels (both P

Remodelling in the atria could appear as a result of hypertension, diabetes or ischaemic heart disease. Galectin-3 (Gal-3) is a mediator of profibrotic pathways and a potential biomarker of cardiac remodelling. We prospectively recruited consecutive patients undergoing elective cardiac surgery. Preoperative Gal-3 levels were determined from serum samples, and the presence of fibrosis was assessed from atrial appendage tissue samples obtained during cardiac surgery. We included 100 patients with aortic valve or ischaemic heart diseases and 15 controls with permanent AF. Gal-3 levels were associated with sex, left atrial volume, previous cardiac disease, diabetes mellitus, hypertension, NYHA and NT-proBNP. We observed differences in serum Gal-3 concentrations between patients and controls with permanent AF (p = 0.020). We performed ROC curves related to fibrosis and established a cutoff point for Gal-3 >13.65 ng/ml. Multivariate analyses showed previous cardiac disease, NYHA scale and high Gal-3 to be independent predictors of fibrosis. After adjustment for confounding factors, atrial fibrosis remained the only independent factor for the development of AF (p = 0.022). High Gal-3 serum levels predict fibrosis of the atrial appendage. NYHA scale and previous cardiac disease were also associated with tissue fibrosis in patients undergoing surgery. Atrial fibrosis was the only independent predictor for post-operative AF occurrence in our model after correcting for confounding factors.

Atrial fibrillation (AF) occurs in ∼ 30% of patients undergoing coronary artery bypass grafting (CABG) and in 40% of patients after valve surgery. High-sensitivity cardiac troponin T (hsTnT) is a specific and high-sensitivity marker of myocardial injury, while N-terminal proB-type natriuretic peptide (NT-proBNP) is an established biomarker for wall remodelling. We investigated whether hsTnT and NT-proBNP levels could be used as valuable biomarkers for AF occurrence after cardiac surgery.We included consecutive haemodynamically stable patients undergoing programmed cardiac surgery with cardiopulmonary bypass pump. We determined hsTnT and NT-proBNP levels before and after cardiac surgery and recorded AF development by prolonged electrocardiogram monitoring.We included 100 patients with predominantly aortic valve (n = 42) or ischaemic heart (n = 58) diseases. Twenty-nine patients (29%) developed post-surgical AF. Patients developing AF had a longer hospital stay (P = 0.005). hsTnT levels increased after surgery [P0.001], indicating perioperative myocardial injury, with higher presurgery levels in patients who developed AF [P = 0.015]. Body mass index and EuroSCORE risk scale were independently associated with higher hsTnT levels presurgery. On univariate analysis, age (P = 0.048), male sex (P = 0.031), indexed left atrial volume (P = 0.042), β-blockers treatment (P = 0.024), type of surgery (valve surgery vs CABG; P = 0.034), EuroSCORE risk scale (P = 0.025) and higher preoperative hsTnT levels (P = 0.009) were predictors of AF development, but NT-proBNP did not reach statistical significance (P = 0.060). hsTnT levels in blood samples obtained the day after surgery were not associated with post-surgical AF development (P = 0.165). In a multivariate model, only higher hsTnT levels before cardiac surgery (11.87 ng/l) [Odds Ratio, OR; (95% Confidence interval, CI) 4.27 (1.43-12.77), P = 0.009] and male sex [OR 5.10 (1.72-15.13), P = 0.003)] were independently associated with the occurrence of post-surgical AF.High presurgical hsTnT levels were independently predictive of patients developing AF after cardiac surgery. hsTnT levels determined post-surgery suggest that cardiac perioperative myocardial injury is not associated with postoperative AF development. NT-proBNP did not reach statistical significance as a biomarker for AF prediction.

Resumen Introduccion y objetivos Las concentraciones basales de interleucina 6 y proteina C reactiva elevadas comportan un aumento del riesgo de muerte en el sindrome coronario agudo sin elevacion del segmento ST. El objetivo del estudio es delucidar si las determinaciones seriadas de interleucina 6 y proteina C reactiva ultrasensible aportan informacion pronostica adicional a las determinaciones basales para la estratificacion del riesgo a largo plazo de los pacientes con sindrome coronario agudo sin elevacion del segmento ST. Metodos Se incluyo prospectivamente en el estudio a 216 pacientes consecutivos con sindrome coronario agudo sin elevacion del segmento ST. Se obtuvieron muestras de sangre en un plazo de 24 h tras el ingreso en el hospital y a los 30 dias de seguimiento. La variable de valoracion principal fue la combinacion de muerte por todas las causas, infarto de miocardio no mortal e insuficiencia cardiaca aguda descompensada. Resultados Las concentraciones tanto de interleucina 6 como de proteina C reactiva ultrasensible se redujeron del dia 1 al dia 30, con independencia de los eventos adversos aparecidos (p Conclusiones En esta poblacion, tanto la concentracion de interleucina 6 como la de proteina C reactiva ultrasensible se reducen tras la fase aguda. La determinacion de las concentraciones de interleucina 6 en muestras seriadas mejora la estratificacion pronostica del riesgo en estos pacientes.

CML cells induce PARP-1 dependent parthanatos in NK cells, and inhibition of PARP-1 maintains lymphocyte viability and function. NK cells and T cells are commonly dysfunctional in CML, and their status may determine the course of disease. We aimed to define the molecular mechanisms of leukemia-induced immunosuppression with focus on the role of ROS and the PARP-1 pathway of cell death. Malignant granulocytes from patients with BCR-ABL-positive CML expressed the oxygen radical-producing enzyme NOX, produced large amounts of ROS, and triggered extensive cell death in NK cells. Inhibition of PARP-1 maintained NK cell viability in cocultures with suppressive leukemic cells. Under conditions of oxidative stress, PARP-1 inhibition upheld the capacity of NK cells to kill myeloid leukemic cells, in addition to restoring the proliferation and cytokine production of NK cells and cytotoxic T cells. Our findings are suggestive of a novel pathway of relevance to immunosuppression in CML.

BackgroundHypertrophic cardiomyopathy (HCM) is characterized by cardiomyocyte hypertrophy and fibrosis. Although is an autosomal dominant trait, a group of nonsarcomeric genes have been postulated as modifiers of the phenotypic heterogeneity. Material and methodsWe prospectively recruited 168 HCM patients and 136 healthy controls from three referral centres. Patients and controls were clinically stable at entry in the study. Nine polymorphisms previously associated with ventricular remodelling were determined: I/D ACE, AGTR1(A1666C), CYP11B2(C344T), PGC1-(G482S), COLIA1(G2046T), ADRB1(R389G), NOS3(G894T), RETN(-420C>G) and CALM3(-34T>A). Their potential influence on prognosis, assessed by hospital admissions, and their cause were recorded. ResultsThe median follow-up time was 495 months. Allele and genotype frequencies did not differ between patients and controls. Thirty-six patients (215%) required urgent hospitalization (185% for heart failure, 222% for atrial arrhythmias, 111% for ventricular arrhythmias, 296% for ischaemic heart disease, 148% for stroke and 37% for other reasons) with a hospitalization rate of 875% per year. Multivariate analysis showed an independent predictive value for noncarriers of polymorphic COL1A1 allele [HR: 276(126-605), P = 0011] and a trend in homozygous carriers of ADRB1 Arg389 variant [HR: 198(099-402); P = 0057]. ConclusionOur study suggests that COL1A1 polymorphism (2046G>T) is an independent predictor of prognosis in HCM patients supporting the importance of nonsarcomeric genes on clinical prognosis in HCM.

Acute myeloid leukemia (AML) is the most common acute leukemia in adults. Although most patients achieve complete remission (CR) after chemotherapy, the majority suffer from subsequent leukemic relapse, which is associated with poor long-term survival. Thus, new therapies to maintain CR are highly warranted. After the completion of chemotherapy, AML patients have a minimal burden of leukemic cells, which are reportedly susceptible to cytotoxic lymphocytes such as NK cells and T cells. A therapy that boosts the function of these effector cells therefore has the potential to eradicate the malignant clone in AML and prevent relapse, Here, we briefly review the literature on the role of the immune system in AML and introduce the rationale for the use of histamine dihydrochloride (HDC) in conjuction with low-dose IL-2 as relapse-preventive immunotherapy for this disease.

Dendritic cells (DCs) communicate with nonadaptive and adaptive lymphocytes on multiple levels. Efficient DC-lymphocyte interactions require that lymphocytes remain viable and functional also under conditions of oxidative stress, such as in microbial infection or in the malignant microenvironment. For this study, we exposed human T and NK cells to oxidants delivered either by autologous phagocytes or in the form of exogenous hydrogen peroxide. In accordance with earlier studies, these lymphocytes became dysfunctional and subsequently apoptotic. The presence of myeloid DCs efficiently rescued T cells (CD4+ and CD8+) and NK cells from oxidant-induced inactivation and apoptosis. The mechanism of the myeloid DC-mediated lymphocyte protection was, at least in part, explained by the capacity of the myeloid DCs to neutralize extracellular oxygen radicals, which, in turn, was reversible upon coincubation with a catalase inhibitor. Our results are suggestive of a novel aspect of DC-lymphocyte interaction that may have implications for lymphocyte function in inflamed tissue.

AimsHypertrophic cardiomyopathy (HCM) is characterized by left ventricular hypertrophy and fibrosis. HCM is an autosomal-dominant disease caused by more than 400 mutations in sarcomeric genes. Changes in nonsarcomeric genes contribute to its phenotypic heterogeneity. Cardiac fibrosis can be studied using late gadolinium enhancement (LGE) cardiac magnetic resonance imaging. We evaluated the potential role of two polymorphisms in nonsarcomeric genes on interstitial fibrosis in HCM. Materials and methodsTwo polymorphisms in nonsarcomeric genes [ACE (deletion of 287bp in the 16th intron) and RETN (-420C>G)] were analysed in 146 HCM patients. Cardiac fibrosis was assessed using LGE to determine the number of affected segments. ResultsAllelic frequencies in ACE and RETN polymorphisms were consistent with the Hardy-Weinbergequilibrium (both P>0.05). We found that the presence of the polymorphic allele in the -420C>G RETN polymorphism was independently associated with the number of affected segments of LGE (P=0.038). Increased circulating resistin concentration, measured by enzyme-linked immunosorbent assay, was associated with a higher degree of cardiac fibrosis. Myocardial fibrosis, assessed by Masson's trichrome staining, was associated with the -420C>G RETN polymorphism in 46 tissue samples obtained by septal myectomy (P=0.044). ConclusionsThe -420C>G RETN polymorphism was independently associated with the degree of cardiac fibrosis, assessed by LGE, in patients with HCM. In addition, there was an association between the polymorphism and the circulating resistin levels as well as with myocardial fibrosis in tissues obtained by myectomy. Investigating the physiological implication of the RETN polymorphism in HCM in combination with the use of imaging technologies might help to establish the severity of disease in patients with HCM.

Cytotoxic T cells and NK cells will acquire features of apoptosis when exposed to oxygen radicals, but the molecular mechanisms underlying this phenomenon are incompletely understood. We have investigated the role of two enzyme systems responsible for execution of cell death, caspases and the poly(ADP-ribose) polymerase (PARP). We report that although human cytotoxic lymphocytes were only marginally protected by caspase inhibitors, PARP inhibitors completely protected lymphocytes from radical-induced apoptosis and restored their cytotoxic function. The radical-induced, PARP-dependent cell death was accompanied by nuclear accumulation of apoptosis-inducing factor and a characteristic pattern of large-fragment DNA degradation. It is concluded that the PARP/apoptosis-inducing factor axis is critically involved in oxygen radical-induced apoptosis in cytotoxic lymphocytes.

Plasma from 173 patients with HCV genotype 1 infection was analyzed for IP-10 levels prior to treatment with pegylated interferon-α-2a and ribavirin. Significantly lower IP-10 levels were observed in patients achieving a rapid viral response (RVR) (P < .0001), even in those with body mass index (BMI) ≥ 25 kg/m2 (P = .004) and with baseline viral load ≥ 2 million IU/mL (P = .001). Similarly, significantly lower IP-10 levels were observed in patients obtaining a sustained viral response (SVR) (P = .0002), including those having higher BMI (P < .05), higher viral load (P = .0005), and both higher BMI and viral load (P < .03). In multivariate logistic regression analyses, a low IP-10 value was independently predictive of both RVR and SVR. A baseline cutoff IP-10 value of 600 pg/mL yielded a negative predictive value (NPV) of 79% (19/24) for all genotype 1–infected patients, which was comparable with that observed using a reduction in HCV-RNA by at least 2 logs after 12 weeks of therapy (NPV 86%; 19/22); by combining the two, 30 of 38 patients (NPV 79%) potentially could have been spared unnecessary therapy. In patients having both higher BMI and viral load, cut-off levels of 150 and 600 pg/mL yielded a positive predictive value (PPV) of 71% and NPV of 100%, respectively. In conclusion, pretreatment IP-10 levels predict RVR and SVR in patients infected with HCV genotype 1, even in those with higher BMI and viral load. A substantial proportion of the latter patients may achieve SVR in spite of unfavorable baseline characteristics if their pretreatment IP-10 level is low. Thus, pretreatment IP-10 analysis may prove helpful in decision-making regarding pharmaceutical intervention. (HEPATOLOGY 2006;44:1617–1625.)

The persistent infection caused by hep- atitis C virus (HCV) is presumably explained by a deficient immune response to the infection, but the basis for the inefficiency of immune-mediated virus eradication is not known in detail. This study ad- dresses mechanisms of relevance to dysfunction of cytotoxic lymphocytes in HCV infection, with a focus on the role of phagocyte-derived oxygen rad- icals. We show that NS3, a nonstructural, HCV- encoded protein, induces a prolonged release of oxygen radicals from mononuclear and polymor- phnuclear phagocytes by activating a key enzyme in radical formation, the reduced nicotinamide ad- enine dinucleotide phosphate (NADPH) oxidase. The NS3-activated phagocytes, in turn, induced dysfunction and/or apoptosis in three major subsets of lymphocytes of relevance to defense against HCV infection: CD3/56 - T cells, CD3 - /56 nat- ural killer (NK) cells, and CD3/56 NKT cells. Two inhibitors of the NADPH oxidase, histamine and diphenylene iodonium, suppressed the NS3- induced oxygen radical production and efficiently protected lymphocytes against NS3-induced apo- ptosis and dysfunction. In conclusion, we propose that NS3, by triggering oxygen radical formation in phagocytes, may contribute to the dysfunction of antiviral lymphocytes in HCV-infected liver tissue and that strategies to circumvent oxidative stress may be useful in preventing HCV-associated carci- nogenesis and facilitating lymphocyte-mediated clearance of infected cells. J. Leukoc. Biol. 76: 1180-1186; 2004.

A significant part of the therapeutic benefit of interferon-α (IFN-α) therapy in malignant diseases and in chronic viral infections is assumed to result from activation of lymphocytes with natural killer (NK) and T cell phenotype. In tumor tissue and in chronically infected tissue, the function and viability of these lymphocytes are frequently impaired. Mononuclear phagocyte (MP)-derived reactive oxygen species (ROS) have been proposed to contribute to the lymphocyte suppression in these tissues. Here, we report that three types of human cytotoxic lymphocytes of relevance to immunoactivation by IFN-α, CD3ɛ+/8+/56– T cells, CD3ɛ–/56+ NK cells, and CD3ɛ+/56+ NK/T cells became anergic to IFN-α induction of the cell-surface activation marker CD69 after exposure to autologous MPs in vitro. In addition to their incapacity to express CD69, cytotoxic lymphocytes acquired features characteristic of apoptosis after incubation with MPs. The lymphocyte apoptosis and nonresponsiveness to IFN-α were prevented by two inhibitors of reduced nicotinamide adenine dinucleotide phosphate oxidase-dependent formation of ROS in MPs, histamine dihydrochloride and diphenylene ionodonium, as well as by catalase, a scavenger of ROS. We conclude that MP-derived ROS may negatively affect IFN-α-induced immunostimulation and propose that ROS inhibitors or scavengers may be useful to improve lymphocyte activation during treatment with IFN-α.

Background Chronic Kidney Disease Epidemiology Collaboration (CKD-EPI) equations estimate glomerular filtration rate more accurately than the Modification of Diet in Renal Disease (MDRD) Study equation. Our aim was to evaluate whether CKD-EPI equations based on serum creatinine and/or cystatin C (CysC) predict risk for major bleeding (MB) more accurately than the MDRD Study equation in patients with non-ST-segment elevation acute coronary syndromes (ACS). Materials and methods Three hundred and fifty consecutive subjects with non-ST-segment elevation ACS (68 ± 12 years, 70% male) were studied. Glomerular filtration rate was estimated using the CKD-EPI and MDRD Study equations. The primary endpoint was the occurrence of MB during the follow-up, which was defined according to the Bleeding Academic Research Consortium Definition criteria as bleeding types 3–5. Results During the median follow-up of 589 days (interquartile range, 390–986), 27 patients had MB (0·04% events per person year). Patients with MB had worse kidney function parameters, regardless of the estimating equation used (P

Beyond cancer-cell intrinsic factors, the immune status of the host has a prognostic impact on patients with cancer and influences the effects of conventional chemotherapies. Metastatic melanoma is intrinsically immunogenic, thereby facilitating the search for immune biomarkers of clinical responses to cytotoxic agents. Here, we show that a multi-tyrosine kinase inhibitor, sorafenib, upregulates interleukin (IL)-15Rα in vitro and in vivo in patients with melanoma, and in conjunction with natural killer (NK) group 2D (NKG2D) ligands, contributes to the Th1 polarization and accumulation of peripheral CD4+NKG2D+ T cells. Hence, the increase of blood CD4+NKG2D+ T cells after two cycles of sorafenib (combined with temozolomide) was associated with prolonged survival in a prospective phase I/II trial enrolling 63 patients with metastatic melanoma who did not receive vemurafenib nor immune checkpoint-blocking antibodies. In contrast, in metastatic melanoma patients treated with classical treatment modalities, this CD4+NKG2D+ subset failed to correlate with prognosis. These findings indicate that sorafenib may be used as an "adjuvant" molecule capable of inducing or restoring IL-15Rα/IL-15 in tumors expressing MHCclass I-related chain A/B (MICA/B) and on circulating monocytes of responding patients, hereby contributing to the bioactivity of NKG2D+ Th1 cells., peer-reviewed

Atrial fibrillation (AF) has an incidence rate of approximately 30% and is the most frequent arrhythmia following heart surgery. Factors such as inflammation, the presence of heart fibrosis, stress and cardiomyocyte apoptosis, have all been associated with AF.We believe that atrial remodelling is a pre-existent process in patients with post-surgical AF. We have analyzed the factors related to the incidence of atrial fibrillation in the period after heart surgery.We included consecutive, hemodynamically stable patients with a sinusal rhythm who were subjected to programmed heart surgery with extracorporeal circulation. An assessment was made of the fall in atrial fibrillation after surgery using prolonged electrocardiographic monitoring.A total of 100 patients were included in the study and were subjected to either coronary revascularisation surgery (59) or aortic valve substitution due to severe aortic stenosis (41). Postoperative AF occurred in 29 patients who received predominantly more valve surgery than coronary surgery. The following factors were predictive of postoperative AF in the multivariate analysis: Male sex; beta-blocker therapy for chronic disease; the use of intraoperative; fibrinogen perfusion; low HDL cholesterol values; and high sensitive troponin T values, in the preoperative period.HDL cholesterol and high sensitive troponin T can be useful biomarkers to predict the occurrence of AF after surgery. The early identification of these patients who develop of FA allows us to take preventive measures to minimize the negative effects.

The resveratrol monomers (trans-piceid, cis-piceid, trans-resveratrol, and cis-resveratrol) have been previously identified and quantified in red wines. Here, the levels of these compounds in Spanish white and rose wines are described. For white wines, a concentration step was needed to quantify the four resveratrol isomers. The recovery and reproducibility of this procedure were very good. These white wines had levels between 0.051 and 1.801 mg/L. Rose wines had an average level of 2.15 mg/L, in between those of red and white wines. Keywords: Resveratrol; piceid; glucosides; white wine; rose wine

A B S T R A C T Introduction and objectives: High baseline levels of interleukin-6 and C-reactive protein confer an increased risk of mortality in non-ST-segment elevation acute coronary syndrome. The aim of the study was to determine whether serial measurements of interleukin-6 and high-sensitivity C-reactive protein provide additional information to baseline measurements for risk stratification of non-ST-segment elevation acute coronary syndrome. Methods: Two hundred and sixteen consecutive patients with non-ST-segment elevation acute coronary syndrome were prospectively included. Blood samples were obtained within 24 h of hospital admission and at 30 days of follow-up. The endpoint was a composite of all-cause death, nonfatal myocardial infarction, or acute decompensated heart failure. Results: Both interleukin-6 and high-sensitivity C-reactive protein levels decreased from day 1 to day 30, regardless of adverse events (both P

Beta-trace protein (BTP) is a low-molecular mass protein belonging to the lipocalin protein family, which is more sensitive than serum creatinine for detecting impaired renal function. The aims of the present study were to evaluate whether plasma BTP improves the risk stratification of patients with non-ST-segment elevation acute coronary syndromes and to compare it to cystatin C (CysC), serum creatinine, and estimated glomerular filtration rate. Two hundred twenty-six consecutive patients with non-ST-segment elevation acute coronary syndromes were prospectively included. Blood samples were obtained within 24 hours of hospital admission to measure BTP, CysC, and creatinine. The study end point was all-cause death. Over a median follow-up period of 859 days (interquartile range [IQR] 524 to 1,164), 24 patients (10.6%) died. Decedents had higher concentrations of BTP (1.03 mg/L [IQR 0.89 to 1.43] vs 0.74 mg/L [IQR 0.61 to 0.92], p0.001), CysC (1.16 mg/L [IQR 0.91 to 1.59] vs 0.90 mg/L [IQR 0.76 to 1.08], p = 0.001), and serum creatinine (1.10 mg/L [IQR 0.87 to 1.46] vs 0.94 mg/L [IQR 0.80 to 1.10], p = 0.004) and a lower mean estimated glomerular filtration rate (60 ± 20 vs 80 ± 24 ml/min/1.73 m(2), p0.001). After multivariate adjustment, BTP and CysC were predictors of all-cause death, while estimated glomerular filtration rate and serum creatinine concentrations did not achieve statistical significance. In stratified analyses according to kidney function, elevated BTP and CysC were associated with a higher risk for all-cause death. Reclassification analyses showed that BTP and CysC added complementary information to Global Registry for Acute Coronary Events (GRACE) risk score. In conclusion, BTP and CysC levels were associated with all-cause death risk and modestly improved prognostic discrimination beyond the GRACE risk score in patients with non-ST segment elevation acute coronary syndromes.

The initial chemotherapy in acute myeloid leukaemia (AML) comprises a first phase of induction and a second phase of consolidation. In the majority of patients, the induction treatment leads to complete remission (CR), defined as microscopic disappearance of leukaemic disease along with the return of normal haematopoiesis. However, despite the introduction of more efficacious consolidation regimens, a worryingly large proportion of AML patients in CR will subsequently experience relapses with poor prospects of long-term survival. A relapse is assumed to be the result of expansion of residual leukaemic cells that have escaped the initial chemotherapy. The anti-leukaemic functions of T cells and natural killer (NK) cells has formed the background to the use of interleukin-2 (IL-2), a T- and NK cell-activating cytokine, with the aim to eliminate residual leukaemia and hence reduce the relapse rate in AML, but the clinical trials using IL-2 monotherapy have yielded disappointment. A recent phase III study has demonstrated that post-consolidation treatment with the combination of histamine dihydrochloride (HDC) and IL-2 significantly prevents relapse in AML patients. Here we account for the preclinical background to the use of HDC/IL-2 in AML along with a review of clinical results.

Phagocyte-derived reactive oxygen species (“oxygen radicals”) have been ascribed a suppressive role in immunoregulation by inducing dysfunction and apoptotic cell death in lymphocytes. Earlier studies show that human NK cells are exceptionally sensitive to oxygen radical-induced apoptosis and functional inhibition. Two subsets of human CD56+ NK cells have been identified: the highly cytotoxic CD56dim cells which constitute >90% of NK cells in peripheral blood, and the less cytotoxic but efficiently cytokine-producing CD56bright cells. In this study, we demonstrate that the CD56bright subset of NK cells, in contrast to CD56dim cells, remains viable and functionally intact after exposure to phagocyte-derived or exogenously added oxygen radicals. The resistance of CD56bright cells to oxidative stress was accompanied by a high capacity of neutralizing exogenous hydrogen peroxide, and by a high cell-surface expression of antioxidative thiols. Our results imply that CD56bright NK cells are endowed with an efficient antioxidative defense system that protects them from oxygen radical-induced inactivation.

The primary objective of this phase 3 study was to determine whether postconsolidation immunotherapy with interleukin-2 (IL-2) and histamine dihydrochloride (HDC) improved the leukemia-free survival (LFS) of adult patients with acute myeloid leukemia (AML) in complete remission (CR). Three hundred twenty patients with AML (median age, 57 years; range, 18-84 years) were stratified by CR1 or subsequent CR (CR1) and randomly assigned to treatment with HDC/IL-2 or no treatment (control). Treatment comprised 10 21-day cycles with IL-2 (16 400 U/kg) plus HDC (0.5 mg); both compounds were administered by subcutaneous injection twice daily. Study arms were balanced for age, sex, previous treatment, leukemic karyotypes, time from CR to inclusion, and frequency of secondary leukemia. Three years after enrollment of the last patient, treatment with HDC/IL-2 was found to improve LFS over control in the study population (CR1 + CR1, n = 320; P.01, log-rank test). For patients in CR1 (n = 261), treatment significantly improved LFS (P = .01) with 3-year LFS estimates of 40% (HDC/IL-2) compared with 26% (control). Side effects were typically mild to moderate. These results indicate that HDC/IL-2 treatment offers an efficacious and tolerable treatment for patients with AML in remission.

Phenols have antioxidant properties, owed by their phenolic hydroxyls, which seem to be responsible for the physiological effects attributed to moderate wine consumption [5,8]. In red vinification, since the musts are macerated with the skins, red wines have a higher content of phenolic compounds. Nevertheless, the phenolics present in white wines seem to be more active than those in red wines [18]. It is important to profile the phenolic compounds present in white wines responsible for this effect. First, we studied resveratrol and piceid isomers [16], because the trans forms are the major compounds in the root of Polyganum cuspidatum, used in traditional oriental medicine for several purposes, among them the treatment of cardiovascular diseases and cancer. These compounds inhibit LDL oxidation [4], block platelet aggregation [2,13], inhibit eicosanoid synthesis [9,13] and show anticancer activity [6]. Different groups of phenolics are present in white wines [5,17]. Due to the importance of these compounds, we attempted to try to identify and quantify the maximum number of these compounds since any one of them may act as an antioxidant. Besides stilbenes (resveratrol and piceid), the presence of 31 phenolic compounds has been studied in white wines, including phenolic acids, trans and cis hydroxycinnamic acids and derivatives, flavan-3-ols, flavonols, and the oligomeric procyanidins, by HPLC [3]. Here, we report the concentration of stilbenes and phenolics present in white wines.

The cytotoxicity of natural killer (NK) cells is dependent on the interaction between target cell ligands and a series of stimulatory receptors on NK cells. Two of these triggering receptors, the NKp46 natural cytotoxicity receptor (NKp46) and the major histocompatibility complex (MHC) class I-interactive NKG2D receptor, are deficiently expressed by NK cells recovered from patients with acute myeloid leukaemia (AML), but little is known regarding the regulation of NKp46 and NKG2D expression. Here we report that mononuclear and polymorphonuclear phagocytes downregulate the cell surface density of NKp46 and NKG2D on NK cells with CD56(dim) phenotype in vitro by a mechanism that is dependent on the availability of phagocyte-derived reactive oxygen species (ROS). Histamine maintained NKp46 and NKG2D expression despite the presence of inhibitory phagocytes by targeting an H2 receptor on phagocytes. By contrast, NKp46 and NKG2D expression by the CD56(bright) subset of NK cells was resistant to inhibition by phagocytes. Our findings are suggestive of a novel mechanism of relevance to the regulation of NKp46/NKG2D receptor expression. Moreover, our findings suggest that the previously reported action of histamine on NK cell-mediated killing of leukaemic cells may be related to the preservation of activatory NK-cell receptors.

Aims: To evaluate whether Beta trace protein (BTP) and cystatin C (CysC) provide information to the CRUSADE bleeding score for predicting major bleeding (MB); and to compare them to other renal function parameters in non-ST-segment elevation acute coronary syndromes (NSTE-ACS). Methods and results: We included 273 pts with NSTE-ACS. NSTE-ACS was defined as ischemic symptoms lasting ≥10 min and occurring ≤72-h before admission and either ST-segment deviation of ≥1 mm or elevated levels of a cardiac biomarker of necrosis. All blood samples were obtained before coronary angiography within 24-h of admission. The study endpoint was MB (BARC definition criteria: bleeding type 3 to type 5). During a follow-up of 760 days [411-1098], 25 pts (9.2%) had MB. Pts with MB had higher BTP (0.98 [0.71-1.16] vs 0.72 mg/L [0.60-0.91], p=0.002), CysC (1.05 [0.91-1.30] vs 0.90 mg/L [0.75-1.08], p=0.003) and lower eGFR (66±27 vs 80±30 mL/min/1.73m2, p=0.02) than pts without MB; there were no differences in creatinine levels between both groups (p=0.14). In multivariate analyses, a BTP>0.97 mg/L (HR=3.4, 95% CI=1.5-7.9, p=0.004) and CysC>0.96 mg/L (HR=3.1, 95% CI=1.2-7.5, p=0.02) were significant predictors of MB, while eGFR using MDRD equations and creatinine were not. Among subjects with eGFR>60ml/min/1.73m2, those with elevated BTP or CysC had a significantly higher risk for MB (Fig. 1). NRI from the addition of BTP and CysC to CRUSADE were 38% and 21% respectively, while the relative IDI were 12% and 4%. ![Figure][1] Figure 1A, B Conclusion: BTP and CysC were superior to conventional renal parameters for predicting MB, and provide complementary information to the CRUSADE score in NSTE-ACS. Future studies should assess the potential role of incorporating these renal function biomarkers into the bleeding risk scales. [1]: pending:yes

Background and Purpose: Natural killer (NK) cells are commonly dysfunctional in chronic myeloid leukemia (CML) and their status may determine the course of disease. We aimed to define the molecular mechanisms of leukemia-induced NK cell inhibition with focus on the role of CML cell-derived reactive oxygen species (ROS) and the poly(ADP-ribose) polymerase-1 (PARP-1) pathway of cell death (parthanatos). Methods: Malignant granulocytes from patients with BCR-ABL+ CML were analyzed using flow cytometry for cell surface expression of the oxygen radical-producing NADPH/NOX-2 oxidase (determined by expression of the NADPH oxidase subunit gp91phox). ROS production by CML cells was assessed by chemiluminescence. In co-culture experiments, purified NK cells from healthy donors were exposed to malignant CML cells followed by analysis of NK cell viability. In these experiments, inhibitors of PARP-1 (PJ34 and EB-47) and the caspase cascade (Z-VAD.fmk) were added to determine the mechanisms of cell death in NK cells exposed to CML cells. Translocation of apoptosis-inducing factor (AIF) from mitochondria to nucleus was determined in sorted NK cells by immunoblot technique. Results: All BCR-ABL+ CML cells showed high cell surface expression of gp91phox, indicating the presence of a NADPH/NOX-2 oxidase. CML cells produced high amounts of ROS upon stimulation with the protein kinase C activator PMA or by the bacterial tripeptide fMLF suggestive of a functional ROS-forming oxidase. Unstimulated CML cells triggered extensive apoptosis-like cell death in co-cultured NK cells, which was prevented by the ROS-degrading enzyme catalase and by the NADPH oxidase/NOX-2 inhibitor diphenylene iodonium. The PARP-1 inhibitor PJ34 (0.5 microM) prevented CML-induced NK cell death at CML/NK cell ratios of 0.125: p Conclusions: Our results imply (1) that paracrine ROS production by CML cells followed by NK cell death may constitute a mechanism by which CML cells evade destruction by cytotoxic lymphocytes, and (2) that CML cells trigger NK cell death by inducing PARP-1 dependent parthanatos. These findings may have implications for the design of more efficient immunotherapeutic protocols in CML. Citation Format: Johan Aurelius, Anna Martner, Rebecca E. Riise, Ana I. Romero, Lars Palmqvist, Mats Brune, Kristoffer Hellstrand, Fredrik B. Thorén. Chronic myeloid leukemic cells trigger poly(ADP-ribose) polymerase-dependent cell death in natural killer cells. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B66.

Oxidative stress, referring to cellular toxicity induced by reactive oxygen metabolites (ROM), has been proposed as a mechanism of immunosuppression in hematological malignant diseases. ROM produced by myeloid cells such as normal or malignant phagocytes have been shown to compromise functions of lymphocytes including natural killer (NK) cells and CD4+ or CD8+ T cells. Phagocyte-derived ROM induce downmodulation of the CD3/CD16 zeta; transduction antigen in these lymphocyte subsets with ensuing loss of function and lack of responsiveness to activating cytokines such as IL- 2. In addition, ROM exposure triggers poly(ADP-ribose) polymerase 1- and apoptosisinducing factor-dependent apoptosis in NK and T cells, but the molecular details of the ROM/lymphocyte interaction are not fully understood. The present study sought to determine the role of the CD16/FcγRIII (CD16) antigen, a signal transduction molecule expressed by NK and T cells and a receptor for the Fc portion of IgG, for oxidative stress in lymphocytes. Human NK (CD56+) or T cells (CD4+ or CD8+) were sorted into pure (>99%) CD16+ or CD16− populations, and incubated overnight with ROM-producing autologous mononuclear phagocytes or exogenous hydrogen peroxide. For all subsets of lymphocytes, the expression of CD16 was a strong determinant of the propensity of cells to acquire features of apoptosis after ROM exposure, as reflected by a pronounced difference of ROM sensitivity between CD16+ and CD16− NK or T cell populations. The higher degree of ROM-induced apoptosis in the CD16+ subsets was observed regardless of whether lymphocytes were incubated with autologous mononuclear phagocytes or exposed to exogenous hydrogen peroxide (p=0.04–0.001 for CD4+/16+, CD8+/16+ and CD16+/56+ lymphocytes as compared with corresponding CD16− subsets). Similar experiments using unsorted T and NK cells suggested that the differential sensitivity to oxidative stress in CD16+ and CD16− lymphocyte subsets was not explained by anti- CD16/CD16 interaction. In conclusion, our findings imply that oxidant-induced apoptosis is preferentially induced in lymphocytes expressing the CD16/FcγRIII antigen.

BACKGROUND: Malignant diseases are frequently accompanied by suppression of cytotoxic lymphocyte function, and a large body of circumstantial evidence suggests that lymphocyte suppression may have implications for disease-free survival in several hematopoietic cancers. In AML, a plethora of immunosuppressive mechanisms have been described including the production of T- and NK-cell-inhibitory factors by AML blasts, a deficient expression of activating receptors on natural killer (NK) cells, and an impaired stimulatory interaction between the CD28 antigen expressed by T cells and contact antigens on AML blasts. We and others have proposed that the production of oxygen radicals (including hydrogen peroxide and its derivatives) by normal and leukemic phagocytic cells may significantly contribute to the state of immunosuppression in AML. The phagocyte-derived oxygen radicals strongly suppress the anti-leukemic cytotoxicity of natural killer (NK) cells with ensuing NK cell death by apoptosis, and inhibitors of oxygen radical formation efficiently rescue NK cells from inhibition and apoptosis. These mechanisms may have implications for AML treatment, as exemplified by the use of the T and NK cell activator interleukin-2 (IL-2) in conjunction with the oxygen radical inhibitor histamine dihydrochloride (HDC) as relapse-protective therapy in AML (Blood108: 88–96). METHODS and RESULTS: The question whether oxygen radical-induced activation apply to CD8+ T cells with spontaneous reactivity against human primary AML blasts has remained unresolved. Autoreactive T cells in AML were recently phenotypically defined as CD3+/8+/45RA+/CCR7 cells (CD8+/45RA+ cells; Blood 100:2132–7). We recovered these cells from human peripheral blood by FACS sorting (FACSAria) with high purity (>98%) and exposed them to exogenous hydrogen peroxide or oxygen radical-producing mononuclear phagocytes. CD8+/45RA+ T cells acquired features of apoptosis at low concentrations of hydrogen peroxide. Also, more than 60% of the CD8+/45RA+ cells acquired features of apoptosis when admixed with autologous mononuclear phagocytes (at a 1:1 phagocyte to T cell ratio, n=11, p CONCLUSIONS: The phenotype of T cells with spontaneous reactivity against AML blasts is highly sensitive to oxidants. By inhibiting oxygen radical formation, HDC efficiently protects these cytotoxic lymphocytes from apoptosis in an environment of oxidative stress. In addition, HDC synergizes with IL-2 to activate and expand CD8+/45RA+ T cells. These mechanisms of T cell activation may be relevant to the prevention of leukemic relapse reported for post-consolidation treatment with HDC/IL-2 in AML.