Mechanism of action of DSP-7888 (adegramotide/nelatimotide) Emulsion, a peptide-based therapeutic cancer vaccine with the potential to turn up the heat on non-immunoreactive tumors

Background Wilms’ tumor 1 (WT1) is highly expressed in various solid tumors and hematologic malignancies. DSP-7888 (adegramotide/nelatimotide) Emulsion is an investigational therapeutic cancer vaccine comprising three synthetic epitopes derived from WT1. We evaluated the mechanism of action of DSP-7888 Emulsion, which is hypothesized to induce WT1-specific cytotoxic T lymphocytes (CTLs) and helper T lymphocytes (HTLs). Methods The ability of nelatimotide and adegramotide to induce WT1-specific CD8+ T cells and CD4+ T cells was assessed in human peripheral blood mononuclear cells (PBMCs). The ability of DSP-7888 Emulsion to induce WT1-specific CTLs in vivo was assessed using human leukocyte antigen-I (HLA-I) transgenic mice. To assess how adegramotide, the helper peptide in DSP-7888 Emulsion, enhances WT1-specific CTLs, HLA-I transgenic mice were administered DSP-7888 or nelatimotide-only Emulsion. Interferon-gamma secretion under antigen stimulation by splenocytes co-cultured with or without tumor cells was then quantified. The effects of combination treatment with DSP-7888 Emulsion and an anti–programmed cell death protein 1 (PD-1) antibody on tumor volume and the frequency of tumor-infiltrating WT1-specific T cells were assessed in HLA-I transgenic mice implanted with WT1 antigen-positive tumors. Result The peptides in DSP-7888 Emulsion were shown to induce WT1-specific CTLs and HTLs in both human PBMCs and HLA-I transgenic mice. Unlike splenocytes from nelatimotide-only Emulsion-treated mice, splenocytes from DSP-7888 Emulsion-treated mice exhibited high levels of interferon-gamma secretion, including when co-cultured with tumor cells (AU)

Weight Loss and Vitamin D Improve Hyporesponsiveness to Corticosteroids in Obese Asthma

Background: Obesity negatively impacts on the response of asthma patients to inhaled corticosteroids. The mechanisms underlying this impact are unknown. Objective: To demonstrate that the poor response to inhaled corticosteroids in obese asthma patients is associated with impaired anti-inflammatory activity of corticosteroids and vitamin D deficiency, both of which are improved by weight loss. Methods: The study population comprised 23 obese asthma patients (OA) (18 females; median (IQR) age 56 [51-59] years), 14 nonobese asthma patients (NOA) (11 females; 53 [43-60] years), 15 obese patients (OP) (13 females; 47 [45-60] years), and 19 healthy controls (HC) (14 females; 43 [34-56] years). Ten OA and 11 OP were evaluated at baseline (V1) and 6 months after bariatric surgery (V2). Corticosteroid response was measured using dexamethasone-induced inhibition of peripheral blood mononuclear cell (PBMC) proliferation. Lung function and serum levels of leptin, adiponectin, and vitamin D were measured at V1 and V2. Results: We found a reduced response to dexamethasone in PBMCs of OP and OA with respect to NOA and HC; this inversely correlated with the adiponectin/leptin ratio and vitamin D levels. Bariatric surgery improved corticosteroid responses in OP and OA and normalized the adiponectin/leptin ratio and vitamin D levels. Exposure of PBMCs to vitamin D potentiated the antiproliferative effects of corticosteroids. Dexamethasone and vitamin D induced similar MKP1 expression in OP and OA. (AU)

Antecedentes: La obesidad tiene un impacto negativo en la respuesta del asma a los corticosteroides inhalados por mecanismos desconocidos. Objetivo: Demostrar que la mala respuesta a los corticosteroides inhalados en pacientes obesos asmáticos se asocia con una actividad antiinflamatoria alterada de los corticosteroides, así como también a la deficiencia de vitamina D, ambos mejorados por la pérdida de peso. Métodos: 23 obesos asmáticos (OA) (18 mujeres; mediana de edad [rango intercuartílico] 56 [51-59] años), 14 asmáticos no obesos (NOA) (11 mujeres; 53 [43-60] años), 15 obesos (O) (13 mujeres; 47 [45-60] años), y 19 controles sanos (HC) (14 mujeres; 43 [34-56] años) fueron incluidos. Se evaluaron 10 pacientes OA y 11 O al inicio (V1) y seis meses después (V2) de cirugía bariátrica. La respuesta a los corticosteroides se midió mediante la inhibición con dexametasona de la proliferación de células mononucleares de sangre periférica (PBMC). La función pulmonar, los niveles séricos de leptina, adiponectina y vitamina D se midieron en V1 y V2. Resultados: Encontramos una respuesta reducida a la dexametasona en PBMC de pacientes O y OA con respecto a los NOA y HC, que se correlacionó de forma inversamente proporcional con la relación adiponectina/leptina y los niveles de vitamina D. La cirugía bariátrica mejoró las respuestas de los corticosteroides en los grupos de pacientes O y OA, y normalizó la relación adiponectina/leptina y los niveles de vitamina D. La exposición de las PBMC a la vitamina D potenció los efectos antiproliferativos de los corticosteroides. La dexametasona y la vitamina D indujeron una expresión similar de MKP-1 en los pacientes O y OA. (AU)

Circulating myeloid-derived suppressors cells correlate with clinicopathological characteristics and outcomes undergoing neoadjuvant chemoimmunotherapy in non-small cell lung cancer

PurposeMyeloid-derived suppressors cells (MDSCs) are heterogeneous immunosuppressive cells, closely related to the development, efficacy and prognosis in various tumors. The relationship between clinicopathological characteristics, efficacy of neoadjuvant chemoimmunotherapy (NCIO) and circulating MDSCs in patients with non-small cell lung cancer (NSCLC) was investigated in this study.MethodsThis study analyzed the clinical data of patients diagnosed at Department of Thoracic Surgery, Beijing Chest Hospital from November 2020 to August 2021. MDSCs and T cells subgroups were measured in fresh peripheral blood mononuclear cells(PBMCs) at baseline. Flow cytometry was used to detect MDSCs and T cells subgroups.ResultsA total of 78 patients with NSCLC and 20 patients with benign nodule underwent direct surgery. 23 patients with NSCLC scheduled to accept NCIO before surgery. NSCLC had elevated levels of total MDSCs, PMN-MDSCs and M-MDSCs compared to patients with benign nodule. MDSCs subgroups were correlated to the pTNM stage in NSCLC patients. The frequency of total MDSCs were moderately positively correlated with regulatory T cells (Tregs)(r = 0.3597, P < 0.01) and negatively correlated with CD4 + T cells(r = 0.2714, P < 0.05). The baseline levels of total MDSCs, PMN-MDSCs and Tregs in pCR patients were significantly decreased than those of non-pCR patients (P < 0.05).ConclusionCirculating MDSCs were increased in NSCLC patients. MDSC subgroups were related to pTNM stage in NSCLC patients. Total MDSCs were positively correlated with Tregs levels and negatively correlated with CD4 + T cells in peripheral blood. The level of MDSCs and Tregs in peripheral blood may have potential value in predicting pathological response in NSCLC.

25-Hydroxyvitamin D potentializes extracellular cathelicidin release from human PBMC stimulated ex vivo with either bacterial (LPS) or viral (P: IC) mimetics

Human cathelicidin refers to the cationic antimicrobial peptide hCAP18/LL-37. LL-37 is formed by cleavage of the propeptide hCAP18 coded by the CAMP gene. The active form of vitamin D, 1,25-dihydroxyvitamin D (1,25(OH)D), has been shown to induce the CAMP gene expression through promoter activation. We previously failed to demonstrate in a clinical trial that supplementation of 25-hydroxyvitamin D (25(OH)D) improves LL-37 serum levels. The aim of this work was to evaluate the impact of 25(OH)D supplementation on intracellular expression of CAMP and secretion of LL-37 in an ex vivo model using the peripheral blood mononuclear cells (PBMC). PBMC collected from healthy donors and incubated with different concentrations of 25(OH)D (0 ng/ml: control (D0); 25 ng/ml: deficient (D25); 75 ng/ml: physiological (D75); 125 ng/ml: supraphysiological (D125)) were stimulated or not with lipopolysaccharide (LPS, 100 ng/ml) or synthetic double-stranded RNA Poly (I: C) (PIC, 10 µg/ml). The intracellular expressions of the CAMP gene and the hCAP18 peptide were measured respectively after 24-h and 48-h incubation periods. The concentration of LL-37 was determined in the culture medium after 48-h incubation. 25(OH)D significantly induced CAMP gene expression at 24 h with a maximum effect at a dose of D125 in either unstimulated (tenfold expression) or stimulated (LPS: 100-fold expression; PIC: 15-fold expression) conditions. Intracellular hCAP18 peptide was overexpressed at 48 h under unstimulated (1.5-fold, D125) and stimulated conditions, LPS (twofold, D125) and PIC (2.5-fold, D125). The secretion of LL-37 in the culture medium was significantly induced by 25(OH)D only in both stimulated (LPS and PIC) conditions in a dose-dependent manner. (AU)

The expression of DNMTs is dramatically decreased in peripheral blood mononuclear cells of male patients with juvenile idiopathic arthritis

INTRODUCTION: Juvenile idiopathic arthritis (JIA) is an autoimmune rheumatic disease, which affects primarily the joints in children under 16 years old. The etiology of JIA is yet unknown but research has shown that JIA is a multifactorial disease implicating several genes and environmental factors. Environmental factors affect immune cells via epigenetic mechanisms. One of the most important epigenetic mechanisms is DNA methylation catalyzed by DNA methyltransferases (DNMTs) and usually associated with gene silencing. In this study, we analyzed the expression of three DNA methyltransferases namely DNMT1, DNMT3a and DNMT3b in peripheral blood mononuclear cells (PBMCs) of patients with JIA and compared it with the expression of these genes in healthy young individuals. MATERIALS AND METHODS: Peripheral blood mononuclear cells of 28 JIA patients and 28 healthy controls were isolated. Total RNA was extracted, cDNA was synthesized and the transcript levels of DNMTs were analyzed by quantitative PCR. RESULTS: Analysis of DNMT1, DNMT3a and DNMT3b relative gene expression in PBMCs of JIA patients and control individuals shows that the expression of DNMT1 and DNMT3a is reduced significantly by 7 folds and 5.5 folds, respectively, in JIA patients compared to healthy controls. Furthermore, the expression of all three DNMTs were significantly and drastically reduced in young affected males compared to healthy males. CONCLUSION: This study shows that the expression of DNMTs is reduced in JIA patients and this reduction is severe in male JIA patients

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Glycoprotein A (GARP) in children who outgrow food allergy

Background: There is little understanding of the mechanisms by which food allergy (FA) develops into persistent disease, or by which symptoms it regresses. Food allergy is a major health problem in developed countries, where the prevalence reaches up to 6% in children and 3% in the adult population. Objective: Children with food allergy remission (FAR) and those without FAR below five years of age, were compared 7-10 years with respect to clinical data and expression of glycoprotein A repetitions predominant (GARP) on peripheral blood mononuclear cells. Methods: Forty children with FAR and 40 children without FAR at age 7-10, in whom FA was previously diagnosed at age below five years were evaluated. In this prospective study, demographic and clinical data were taken, patients were classified as atopic based on history and serum specific IgE (sIgE) for a specific allergen. Blood samples were obtained from all patients to assess expression of GARP. Results: We observed higher expression of GARP in children with FAR compared to children without FA (p = 0.005); optimal cut-off for GARP prediction of the remission was 20.1%. Children with FAR and food-specific IgE in serum had higher expression of GARP compared to children with low food specific IgE (< 0.35 kU/L). Keeping pets at home decreased, and presence of allergic rhinitis increased ORs for high expression of GARP (hGARP) in our patients. Conclusion: hGARP (>20.1%) is related with FAR in school children. Allergic rhinitis, and pets at home modify this effect of GARP. Children with allergic rhinitis have less chance of developing remission despite maintaining immune tolerance (hGARP); quite the opposite case with pets at home

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Effect of ginger extract on expression of GATA3, T-bet and ROR-yt in peripheral blood mononuclear cells of patients with Allergic Asthma

Introduction and objectives: Allergic asthma is a chronic inflammatory disorder of the airways. Th1, Th2 and Th17 cells are the main cells involved in the pathophysiology of asthma. The function of these cells is affected by T-bet, GATA3 and RORgammat transcription factors (respectively). Therefore, the aim of this study was to evaluate the effect of ginger (officinal Roscoe) extract on the expression of T-bet, GATA-3 and ROR-gamma in peripheral blood mononuclear cells (PBMC) of asthmatic patients, in comparison with healthy volunteers as controls. Materials and methods: In this case-control study, a total of 50 individuals including 25 patients with severe, moderate and mild allergic asthma and 25 unrelated healthy controls were involved. The PBMCs were isolated and divided into four groups: negative control, two positive controls (Budesonide and PHA) and ginger-extract treated group. After cell treatment and incubation for 48h, PBMCs were isolated and cDNA was synthesized. Gene expressions of T-bet, GATA3 and ROR-γt were evaluated by Real-time PCR. Results: According to the results of this study, hydroalcoholic extract of ginger could reduce the expression of GATA-3, ROR-gammat, and T-bet in PBMCs of asthmatic patients in comparison with untreated PBMCs (P values = 0.001, 0.001, and 0.002, respectively). It was also shown that the ginger extract could affect T-bet/GATA-3, T-bet/ROR-gamma, and ROR-gammat/GATA-3 expression ratios. Conclusions: This study showed that the use of ginger extract could control asthma and decrease the severity of this disease by affecting the main cells involving the symptoms of asthma in the airways

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Cambios en la expresión de adrenoceptores y GRK en pacientes con insuficiencia pulmonar valvular crónica / Changes in adrenoceptor and GRK expression in patients with chronic pulmonary regurgitation

Introducción y objetivos: La insuficiencia pulmonar (IP) es una complicación frecuente tras la intervención de cardiopatías congénitas. La expresión en leucocitos mononucleares circulantes (LMC) de los adrenoceptores (ß1 y ß2) y de las cinasas (GRK2, GRK3 y GRK5) refleja los cambios neurohumorales que se producen en la insuficiencia cardiaca (IC). El objetivo principal es describir la expresión génica de dichas moléculas en LMC de pacientes con IP grave. Métodos: Estudio prospectivo que analizó la expresión de las moléculas descritas en LMC de pacientes con IP grave en comparación con controles sanos y pacientes con IC avanzada. Resultados: Se estudió a 35 pacientes con IP grave, 22 controles y 13 pacientes con IC. El análisis de comparaciones múltiples mostró que en los controles la cantidad de ARN mensajero de adrenoceptor ß2 era mayor que el que presentaban los pacientes con IP y con IC, con similar expresión en estos 2 grupos: 748,49 (intervalo, 1.703,87) frente a 402,80 (1.210,81) frente a 287,46 (685,69) (p = 0,001). Estos mismos hallazgos se obtuvieron en la expresión génica de GRK2: 760,89 (1.169,46) frente a 445,17 (1.190,69) frente a 284,09 (585,27) (p < 0,001). No hubo diferencias en la expresión de estas moléculas según las variables clínicas de los pacientes con IP. Conclusiones: El patrón de expresión génica de GRK2 y del adrenoceptor ß2 de los pacientes con IP grave, como marcadores moleculares de disfunción cardiaca, se encuentra alterado respecto a los controles y es similar al de los pacientes con IC avanzada

Introduction and objectives: Pulmonary regurgitation (PR) is a frequent complication after repair of congenital heart disease. Lymphocyte expression of adrenoceptors (ß1 and ß2) and kinases (GRK2, GRK3, and GRK5) reflects the neurohumoral changes that occur in heart failure (HF). The main objective of this study was to describe the gene expression of these molecules in circulating lymphocytes in patients with severe PR. Methods: A prospective study was conducted to analyze lymphocyte expression of these molecules in patients with severe PR and compare it with expression in healthy controls and patients with advanced HF. Results: We studied 35 patients with severe PR, 22 healthy controls, and 13 patients with HF. Multiple comparisons analysis showed that ß2-adrenoceptor gene expression levels were higher in the control group than in patients in the PR and HF groups and that expression in the latter 2 groups was similar (748.49 [rank 1703.87] vs 402.80 [rank 1210.81] vs 287.46 [rank 685.69] P = .001). Similar findings were obtained in gene expression of GRK2 (760.89 [rank 1169.46] vs 445.17 [rank 1190.69] vs 284.09 [rank 585.27] P < .001). There were no differences in expression levels of these molecules according to clinical variables in patients with PR. Conclusions: The gene expression pattern of GRK2 and ß2-adrenoceptor as molecular markers of cardiac dysfunction was altered in patients with severe PR compared with controls and was similar to expression in patients with advanced HF

Perspective on immune oncology with liquid biopsy, peripheral blood mononuclear cells, and microbiome with non-invasive biomarkers in cancer patients

Antibodies against immune checkpoint inhibitors such as anti-programmed cell death protein 1 (PD-1) and anti-programmed death ligand 1 (PD-L1) play a key role in the treatment of advanced lung cancer. To examine the clinical benefits of these agents, preclinical and clinical studies have been conducted to identify definitive biomarkers associated with cancer status. Analysis of the blood and feces of tumor patients has attracted attention in recent studies attempting to identify non-invasive biomarkers such as cytokines, soluble PD-L1, peripheral blood mononuclear cells, and gut microbiota. These factors are believed to interact with each other to produce synergistic effects and contribute to the formation of the tumor immune microenvironment through the seven steps of the cancer immunity cycle. The immunogram was first introduced as a novel indicator to define the immunity status of cancer patients. In this review, we discuss the progress in the identification of predictive biomarkers as well as future prospects for anti-PD-1/PD-L1 therapy

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Evidence for the link between defective autophagy and inflammation in peripheral blood mononuclear cells of type 2 diabetic patients

Autophagy was shown to modulate inflammation in immune cells. This study was designed to evaluate the association between autophagy and inflammation in peripheral blood mononuclear cells (PBMCs) of type 2 diabetic (T2D) and non-diabetic (ND) subjects. The autophagy markers were measured by real-time PCR and western blot. The gene expression of pro- and anti-inflammatory cytokines was assessed by real-time PCR. Reduced transcription of BECN1 and LAMP2 and unchanged expression of MAP1LC3B and ATG5 were observed in PBMCs of T2D patients. Decreased LC3B-II and increased p62/SQSTM1 levels were found in PBMCs of diabetic patients. The p-mTOR level was higher in PBMCs of diabetic patients. An increase in both IL-1Beta and TNF-alfa gene expression, along with a decrease in the expression of IL-10, was observed in PBMCs of T2D patients. TNF-α mRNA expression was inversely correlated with the mRNA expression of BECN1 and LAMP2. TNF-alfa and IL-1Beta expression were negatively correlated with the protein levels of LC3B-II. TNF-alfa and IL-1Beta expression had also a positive correlation with protein level of p62. IL-10 mRNA expression was positively correlated with the mRNA expression of BECN1 and LAMP2 and protein levels of LC3B-II and negatively correlated with protein level of p62. In addition, p-mTOR level was positively correlated with IL-1Beta and TNF-alfa mRNA expression. The results revealed a reduced autophagy in PBMCs of T2D patients that is liked with an enhanced inflammation. The suppression of autophagy in PBMCs of diabetic patients may be associated with the activation of the mTOR signaling

Immune response of toddlers with history of prematurity

Background: It is not quite well established how immune responses differ in term and preterm infants beyond the first year of life. This study aimed to evaluate aspects of the innate and adaptive immune responses in a group of preterm infants in comparison with their term peers. Methods: In this cross-sectional study peripheral blood mononuclear cells (PBMC) were isolated from preterm and term children at age three years. Innate immune response was evaluated by the analysis of TLR receptors expression on CD11c+HLADRhigh cells and inflammatory cytokine production after PBMC stimulation with Toll like receptors (TLR) ligands. Adaptive immune response was evaluated by T cells’ phenotyping and function after stimulation with polyclonal conventional T cell stimulus. Conclusion: We have found that the patterns of innate and adaptive immune responses at 3 years of age were not affected by the fact of the children having being born preterm or at term (AU)

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Alternaria alternata acts on human Monocyte-derived Dendritic cells to mediate Th2/Th17 polarisation

INTRODUCTION: Although the mechanism of asthma is not precisely understood in humans, clinical and epidemiological studies have offered a potential relationship between exposure to environmental fungi, such as Alternaria alternata (A. alternata) and the development and exacerbation of asthma. The aim of this project is to investigate the mechanisms of Th2 responses by A. alternata as a clinically relevant model for the environmental exposure. MATERIALS AND METHODS: Plastic adherent monocytes were cultured with granulocyte macrophage colony stimulating factor (GM-CSF) and interleukin-4 (IL-4) to convert these cells into Monocyte-derived Dendritic cells (MoDc) and then matured in the presence of Monocyte-Conditioned Medium (MCM) as the control group and MCM+ A. alternata extract as the inductive groups. RESULTS: The results indicated that the expression of CD14 decreased and CD83 and anti-human leukocyte antigen-DR (HLA-DR) increased in the inductive groups in comparison with the control group. More importantly, A. alternata inhibited IL-12 production by activated dendritic cells (DCs), and the DCs exposed to A. alternata enhanced the Th2 polarisation of CD4+ T cells. The production amount of IL-10 overcame IL-12 as well as Il-23 increased significantly, and hand in T cells the production of cytokines Interferon-γ (IFN-γ) decreased. However, both IL-17 and IL-4 increased (p < 0.05). Phagocytic activity in the inductive groups decreased significantly compared with the control group. CONCLUSION: The asthma-related environmental fungus A. alternata, with an effect on dendritic cells profile mediates TH2/TH17. Such immunodysregulation properties of causative environmental fungi may explain their strong relationship with human asthma and allergic diseases

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IL-4 induces the formation of multinucleated giant cells and expression of Beta5 integrin in central giant cell lesion

BACKGROUND: It is now well established that IL-4 has a central role in the development of monocytes to multinucleated giant cells (MGCs) by inducing the expression of integrins on the surface of monocytes. The aim of this study was to investigate the potential role of IL-4 in induction of β5 integrin expression in the peripheral blood samples of patients with giant cell granuloma. MATERIAL AND METHODS: Monocytes were isolated from peripheral blood samples of patients with central giant cell granuloma (CGCG) and healthy controls using human Monocyte Isolation Kit II. Isolated monocytes were then cultured in the absence or presence of IL-4 (10 and 20 ng/mL), and following RNA extraction and cDNA synthesis, Real-time PCR was performed to determine the level of β5 integrin expression. The formation of CGCGs and morphological analyses were done under light microscopy. For confirmation of CGCGs, immunocytochemistry technique was also carried out by anti-RANK (receptor-activator of NF-κB ligand) antibody. RESULTS: In both patient and control groups, β5 levels were significantly enhanced by increasing the IL-4 dose from 10 to 20 ng/mL. In addition, these differences were significant between patient and control groups without IL-4 treatment. On the other hand, the number of cells which expressed RANK and therefore the number of giant cells were significantly higher in the patient group in comparison to controls, as assessed by immunohistochemistry evaluations. CONCLUSIONS: In this study, we showed an elevation in the expression levels of β5 integrin when stimulated by IL-4. It is strongly indicated that this integrin acts as an important mediator during macrophage to macrophage fusion and development of giant cells

Docosahexaenoic diet supplementation, exercise and temperature affect cytokine production by lipopolysaccharide-stimulated mononuclear cells

Acute exercise induces changes in peripheral mononuclear cells’ (PBMCs) capabilities to produce cytokines. The aim was to investigate the effect of docosahexaenoic acid (DHA) diet supplementation on cytokine production, by lipopolysaccharide (LPS)-stimulated PBMCs after exercise, and the in vitro influence of temperature. Fifteen male soccer players were randomly assigned to a placebo or an experimental group. The experimental group consumed an almond-based beverage enriched with DHA (1.16 g DHA/day) for 8 weeks, whereas the placebo group consumed a similar non-enriched beverage. Blood samples were taken before and after the nutritional intervention in basal conditions and 2 h after acute exercise. Nutritional intervention significantly increased the DHA content in erythrocytes only in experimental group (from 34 ± 3.6 to 43 ± 3.6 nmols DHA/109 erythrocytes). Exercise significantly increased Toll-like receptor 4 (TLR4) in PBMCs but only in the placebo group (203 %). Exercise also significantly increased IL6, IL8, VEGF, INFγ, TNFα, IL1α, IL1β, MCP1, and EGG production rates by LPS-stimulated PBMCs, and this response was attenuated by DHA supplementation. Temperature but not DHA also affected the pattern of cytokine production increasing IL6, IL8, IL1β, and MCP1 synthesis. The higher change was evidenced in IL1β increasing the production rate at 39.5 °C from 3.19 ± 0.77 to 22.4 ± 6.1 pg/h 106 PBMC in placebo and from 2.36 ± 0.11 to 10.6 ± 0.38 pg/h 106 PBMC in the supplemented group. The profile of affected cytokines differs between temperature and exercise, suggesting a different PBMC activation pathway. DHA diet supplementation only attenuated cytokine production after exercise and not that induced by temperatura (AU)

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Evaluación de las modificaciones genéticas que induce la dieta a través de las células mononucleares sanguíneas / Assessment of the genetic modification inducing diet through blood mononuclear cells

El término nutrigenómica fue creado para describir cómo la nutrición afecta a los genes y a las funciones de la proteínas, a nivel transcripcional, proteómico y metabólico. El uso de las modificaciones en la expresión génica en las células mononucleares sanguíneas (CMNS) podría ser un modelo que permita evaluar los estudios de intervención dietética con el objetivo de comprender los mecanismos subyacentes y la influencia de la dieta y los nutrientes en la aterosclerosis, la resistencia a la insulina, la obesidad y la diabetes mellitus. Existen trabajos que han modificado el aporte dietético de colesterol, grasas poliinsaturadas, grasas monoinsaturadas y antioxidantes, y disminuido el aporte calórico, mostrando una gran variedad de efectos sobre la expresión de RNAm en CMNS de genes relacionados con la inflamación, la inmunidad, el metabolismo lípidico, etc. Estos hallazgos moleculares afianzan el conocimiento sobre la respuesta de nuestro organismo a la dieta y abren la posibilidad del análisis rápido de nuevas vías diagnósticas e incluso de nuevas herramientas terapéuticas (AU)

The term nutrigenomics was created to describe how nutrition affects genes and the functions of the protein, at the transcriptional level, proteomic, and metabolic. Using changes in gene expression in blood mononuclear cells could be a model to assess the dietary intervention studies in order to understand the underlying mechanisms and impact of diet and nutrients in atherosclerosis, resistance insulin, obesity and diabetes mellitus. There are studies that have changed the dietary intake of cholesterol, polyunsaturated fat, monounsaturated, antioxidants and decreased caloric intake showing a variety of effects on the expression of mRNA in blood mononuclear cells related to inflammation, immunity, lipid metabolism genes, etc. These molecular findings entrench awareness of our body’s response to diet and open up the possibility of rapid analysis of new diagnostic pathways in this area of knowledge and even new therapeutic tools (AU)

Análisis de factores celulares y moleculares de la frontera tumoral relacionados con la afectación de los ganglios centinela y no centinela en el cáncer de mama / Analysis of cellular and molecular factors at the tumoral margin related to sentinel and non-sentinel lymph node involvement in breast cancer

Objetivos. Investigar el valor de predicción de afectación tumoral del ganglio linfático centinela (GLC) de factores celulares y moleculares determinados en la frontera tumoral de los tumores primarios, así como el valor de esas expresiones en los GLC para predecir la afectación de los ganglios linfáticos no centinela (GLNC) en el cáncer de mama. Pacientes y métodos. Se analizaron muestras tisulares de 59 pacientes que se sometieron a la fase de validación de la técnica de la biopsia selectiva del GLC por cáncer de mama. Se seleccionaron aquellas muestras tisulares de la frontera tumoral de los tumores primarios y de los GLC, sobre los que se realizaron estudios inmunohistoquímicos utilizando mallas de tejido y anticuerpos específicos frente a D2-40, CD3, CD20, CD68, CD138, metaloproteasa (MMP)-1, MMP-7, MMP-13 y inhibidor de metaloproteasas de tejido-1 (TIMP-1). Resultados. La invasión linfática tumoral, el número de células mononucleares inflamatorias (CMI) CD68-positivas o CD138-positivas, se asociaron de forma positiva y significativa con la afectación tumoral de los GLC; mientras que la expresión de TIMP-1, tanto por las células tumorales como por los CMI o fibroblastos del estroma tumoral, se asoció significativamente de forma negativa. La expresión de MMP-1 por las CMI del GLC neoplásico se asoció significativamente con la afectación tumoral de los GLNC. Conclusiones. La determinación de factores celulares y moleculares en la frontera tumoral puede contribuir a conocer mejor las diferentes fases de la metastatización ganglionar linfática en el cáncer de mama (AU)

Objectives. To investigate the predictive value of tumour involvement of the sentinel lymph node (SLN) in terms of specific cellular and molecular factors at the tumoural margin of primary tumours, as well as the value of expression of these factors in the SLNs to predict the involvement of non-SLNs in breast cancer. Patients and methods. Tissue samples from 59 patients who underwent the validation phase of SLN biopsy of breast cancer were analyzed. Tissue samples from the tumoural margin of primary tumours and from SLNs were selected. Immunohistochemical studies were performed using the tissue array technique and antibodies against D2-40, CD3, CD20, CD68, CD138, metalloprotease (MMP)-1, MMP-7, MMP-13 and the tissue inhibitor of metalloproteases-1 (TIMP-1). Results. Tumoural invasion of the lymph nodes and the number of CD68- or CD138-positive mononuclear inflammatory cells (MICs) were positively and significantly associated with tumoural involvement of the SLNs. TIMP-1 expression, both by tumour cells and by MICs or fibroblasts from the tumor stroma, was negatively and significantly associated with tumoural involvement. MMP-1 expression by MICs from neoplastic SLNs was significantly associated with tumoural involvement of non-SLNs. Conclusions. Determination of both cellular and molecular factors at the tumoural margin may contribute to better assessment of the different phases of lymph node metastases in breast cancer (AU)

Peripheral blood mononuclear cells from severe asthmatic children release lower amounts of IL-12 and IL-4 after LPS stimulation

INTRODUCTION: Asthma is an inflammatory disorder of the airways associated with bronchial hyperresponsiveness, airway obstruction, and increased mucus production, with a predominance of type 2 immune response (Th2). According to the hygiene hypothesis, exposure to environmental bacterial lipopolysaccharide (LPS) may induce a type 1 immune response (Th1), modulating the development of asthma. OBJECTIVE: In this study we investigated cytokine production by peripheral blood mononuclear cells (PBMC) from children and adolescents with severe asthma, in response to LPS stimulation in vitro. MATERIALS AND METHODS: 26 children were selected: 13 severe asthmatics and 13 healthy controls, aged between 5 and 18 years. They were evaluated through routine medical history, physical examination and lung function test to diagnose severe asthma. Allergy status was confirmed by skin prick test and specific IgE assay. We collected blood samples to analyse in vitro LPS-induced cytokines release by PBMC. RESULTS: PBMC from severe asthmatic children produced lower levels of IL-12p70 in basal conditions and after 12 and 24 h stimulation with LPS compared to healthy controls. PBMC from severe asthmatic children produced lower levels of IL-4 after 24 h LPS stimulation compared to healthy controls. PBMC from severe asthmatic children produced more levels IL-17 and IL-10 after stimulus with LPS compared to healthy controls. The release of IFN-γ, IL-5 and TNF-α by PBMC from severe asthmatic children was similar to healthy controls. CONCLUSION: Our results demonstrate that LPS directly influence the cytokine profile of PBMC in children with severe asthma. These observations may be potentially helpful in developing new treatment strategies

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Competitive apnea diving sessions induces an adaptative antioxidant response in mononucleated blood cells

The aim was evaluating the effects of hypoxia/reoxygenation repetitive episodes during 5 days of apnea diving (3-day training/2-day competition) on peripheral blood mononuclear cells (PBMCs) antioxidant defenses, oxidative damage, and plasma xanthine oxidase activity. Blood samples, from seven professional apnea divers, were taken under basal conditions the previous morning to the first training session (pre-diving basal), 4 h after ending the competition (4 h post-diving) and the following morning (15 h after last dive) in basal conditions (post-diving basal). Glucose levels significantly decreased whereas triglycerides increased at 4 h post-diving, both returning to basal values at post-diving basal. Glutathione reductase and catalase activity significantly increased after 4 h post-diving remaining elevated at post-diving basal. Glutathione peroxidase and superoxide dismutase activities and catalase protein levels progressively increased after diving with significant differences respect to initial values at post-diving basal. No significant differences were observed in circulating PBMCs and oxidative damage markers. Plasma xanthine oxidase activity and nitrite levels, but not the inducible nitric oxide synthetase, significantly increased 4 h post-diving, returning to the basal values after 15 h. In conclusion, chronic and repetitive episodes of diving apnea during five consecutive days increased plasma xanthine oxidase activity and nitric oxide production which could enhance the signalling role of reactive oxygen and nitrogen species for PBMCs antioxidant adaptation against hypoxia/reoxygenation

La eliminación de células prostáticas circulantes primarias en sangre posterior a la prostatectomía radical disminuye el riesgo de recidiva bioquímica / Elimination of primary circulating prostate cells after radical prostatectomy for prostate cancer decreases the risk of future biochemical failure

OBJETIVO: Las células prostáticas circulantes en sangre (CPCs) primarias son aquellas células prostáticas detectadas en pacientes con cáncer de próstata antes del tratamiento quirúrgico radical, por el contrario, las CPCs secundarias son aquellas detectadas posterior a este tratamiento. Pese a que las CPCs primarias son encontradas frecuentemente en pacientes con cáncer de próstata, solo unas pocas sobreviven y formarán metástasis. Evaluamos la asociación de las CPCs primarias y secundarias con la recidiva bioquímica en hombres con cáncer de próstata tratados con prostatectomía radical. MÉTODOS: Se tomaron muestras de sangre seriadas antes y después del tratamiento quirúrgico, se obtuvieron células mononucleares por centrifugación diferencial y se identificaron las CPCs utilizando inmunocitoquímica. Los datos de edad, estadio patológico, grado patológico, márgenes quirúrgicos, extensión extracapsular, perineural, vascular e infiltración linfática fueron comparados con la presencia o ausencia de CPCs en pacientes con o sin recidiva bioquímica. Se utilizo el método de Kaplan Meyer para comparar la sobrevida libre de enfermedad de los pacientes con o sin CPCs. RESULTADOS: 138 de 423 (32,6%) de los hombres que fueron sometidos a una biopsia prostática por un PSA elevado tuvieron cáncer de próstata, de estos hombres, 15 (10,9%) fueron negativos para CPCs. De los hombres positivos, 95 fueron sometidos a una prostatectomía radical, no existió relación entre la detección de CPCs primarias y los parámetros clínico - patológicos del cáncer, sin embargo, los pacientes con CPCs secundarias se asociaron con mayor tasa de recidiva bioquímica. CONCLUSIONES: Las CPCs primarias son frecuentemente detectadas en hombres con cáncer de próstata, pero no se asocian con recidiva bioquímica, por lo tanto pueden ser útiles para la detección de cáncer de próstata pero no para su pronóstico. La detección de CPCs posterior a la cirugía se asocia con mayores posibilidades de recidiva bioquímica

OBJECTIVES: Primary CPCs are those detected in the blood of prostate cancer patients before radical treatment; secondary CPCs are those detected afterwards. Although primary CPCs are frequently found, it has been suggested that only a few will survive and go on to form metastasis. We evaluate the frequency of primary and secondary CPC detection and the association with biochemical failure, relation with clinical-pathological parameters and clinical implications in men treated by radical prostatectomy (RP) for prostate cancer. METHODS: Serial blood samples were taken before surgery and during follow up after RP. Mononuclear cells were obtained by differential gel centrifugation, and CPCs were identified using standard immunocytochemistry using anti-PSA monoclonal antibodies. Age, pathological stage (organ confined, non organ confined), pathological grade, margin status (positive, negative), extracapsular extension, perineural, vascular, and lymphatic infiltration (positive, negative) were compared with the presence/absence of CPCs in patients with and without biochemical failure. Kaplan Meier method was used to compare the unadjusted biochemical failure free survival of patients with and without CPCs. RESULTS: 138 of 423 (32.6%) men undergoing prostate biopsy for an elevated serum PSA were diagnosed of prostate cancer. Of these men 15 (10.9%) were CPC negative. 95 CPC positive men underwent RP. There was no relation between primary CPC detection and clinical-pathological parameters; however, secondary CPCs were associated both with clinical-pathological parameters and biochemical failure. CONCLUSIONS: Primary CPCs are frequently detected in men with prostate cancer, but they are not associated with biochemical failure, so that they may be useful for prostate cancer detection but not for prognosis. The persistence of CPCs after surgery is associated with increased biochemical failure