Modulación de marcadores de inflamación y estrés oxidativo en pacientes con diabetes tipo 1 y sobrepeso que participan en un programa de cambio de estilo de vida / Modulation of Inflammatory and oxidative stress markers in overweight type 1 Diabetes patients participating in a lifestyle change program

Introduction: Reduction in the expression of inflammatory markers and oxidative stress associated with exercise will protect against cardiovascular complications in Diabetes Mellitus (DM). Aim: The aim of this study was evaluated cardiovascular fitness (VO2 Max), interleukin-6 (IL-6), monocyte chemo-attractant protein 1 (MCP-1) and serum lipid peroxidation (TBARS) in overweight patients with Type-1 diabetes (T1DM) participating in a lifestyle-change program. Results: 20 T1DM overweight patients (43.3 ± 13.8 years), with BMI= 29.6 ± 3.5 kg/m2 , initial HbA1c 7.9 ± 0.91% and treated with multiple insulin injections, were included in this work. The lifestyle-change program consisted of: a) walking 10,000 steps/day, b) sequence of exercises of 24 minutes, 3-5 times/week, c) ¨healthy-plate¨ (and counting carbohydrates, and d) prandial insulin as blood-glucose levels. VO2 max, HbA1c, TBARS, IL6, MCP-1 were determined before starting the lifestyle-change program. Six months of adherence later, participants showed an average number of steps of 8242 ± 1834, a significant increase in VO2 max, (33.4 ±1.3 vs 36.2 ±1.5 ml.Kg-1.min-1 p= 0.008), a significant decrease in serum MCP-1 (314 ±42 vs 235 ±43 MFI p= 0.02), and less TBARS (3.01 ±0.44 vs 2.12 ±0.22 µmol/mL p= 0.015). IL-6 and HbA1c showed no significant decrease. Conclusion: Our results showed that a 6-month systemized and simple exercise plan improves cardiorespiratory fitness (VO2 max), and reduces both circulating oxidative stress and inflammation markers in overweight patients with T1DM.

Introducción: La reducción en la expresión de marcadores inflamatorios y de estrés oxidativo asociado con el ejercicio podría proteger contra las complicaciones cardiovasculares de la diabetes mellitus (DM). Objetivo: El objetivo de este estudio fue evaluar en pacientes con DM tipo1 (DMT1) y sobrepeso, la capacidad cardiorespiratoria (VO2 Max), la expresión sérica de marcadores inflamatorios (IL-6 y MCP-1) y la peroxidación lipídica sérica (TBARS), luego de participar por 6 meses de un programa de cambios de estilo de vida. Resultados: Veinte pacientes adultos (43.3 ± 13.8 años), de ambos sexos, con un Índice de Masa Corporal de 29.6 ± 3.5 kg/m2 , HbA1c inicial de 7,9% ± 0,91, en tratamiento con inyecciones múltiples de insulina participaron del estudio. Se indicó: 1) caminar 10.000 pasos/día, 2) realizar en domicilio una secuencia de ejercicios de 20 minutos, 3-5 veces/semana, 3) plato saludable (consumo de 1 fruta antes de las 3 comidas principales), 4) Insulina prandial según glucemia y conteo de carbohidratos. Se registraron parámetros antropométricos, presión arterial, se determinó VO2 max, y se midieron los niveles séricos de HbA1c, IL6, MCP-1 y TBARs. Luego de seis meses, los participantes alcanzaron un número promedio de pasos de 8242 ± 1834 y mostraron un aumento significativo en VO2 max, (33.4 ±1.3 vs 36.2 ±1.5 ml.Kg-1.min-1 p= 0.008). Además, se encontró una disminución significativa de MCP-1 (314 ±42 vs 235 ±43 MFI p=0.02) y TBARs (3.01 ±0.44 vs 2.12 ±0.22 µmol/mL p= 0.015) en comparación con el día 0. No se observaron modificaciones en los niveles de IL-6 y HbA1c. Conclusión: Nuestros resultados demuestran que el ejercicio, implementado como un plan accesible y acompañado, es adecuado para reducir los riesgos de inflamación y estado pro-oxidativo en pacientes con DM tipo1.

relationship between some endometrial secretion cytokines and in vitro fertilization

Endometrial secretion analysis is a non-invasive and promising method in evaluation of endometrial receptivity. The aim of the present study was to assess the relationship between the success rate of IVF procedures and some endometrial secretion cytokines, including interleukin-1beta [IL-1beta], tumor necrosis factor [TNF-alpha], interferon gamma-induced protein 10 [IP-10], and monocyte chemoattractant protein [MCP]. In a prospective cohort study, 50 women selected for IVF met the study inclusion criteria. All the patients underwent endometrial secretion aspiration prior to embryo transfer. The level of IL-1beta, TNF-alpha, IP-10 and MCP were analyzed by enzyme-linked immunosorbent assay method using special standard kits. To detect successful implantation and pregnancy patients underwent serum human chorionic gonadotropin measurements and ultrasound evaluation. Five samples were excluded. Nine women [20%] had successful clinical pregnancies, which resulted in live birth. Other 36 women [80%] were classified as failed pregnancy. Comparison of cytokine levels showed lower concentrations of TNF-alpha, IP-10, and MCP in the group with successful clinical pregnancy compared to the group with failed pregnancy [p=0.007, 0.005 and 0.001, respectively]. However, no significant difference was revealed in IL-1beta levels between two groups [p=0.614]. The current study suggested that lower concentrations of TNF-alpha, IP- 10, and MCP in endometrial secretions might be associated with improved endometrial receptivity and IVF outcome. Regarding IL-1beta, no statistically significant differences were seen between the groups with and without successful pregnancy

Gene - Gene Interactions Among MCP Genes Polymorphisms in Asthma

PURPOSE: Monocyte chemoattractant proteins (MCPs) are important cytokines that involved in cellular activation and releasing of inflammatoy mediators by basophils and eosinophils in allergic disease. Some MCP gene variants implicate in asthma and monoclonal antibody for MCP-3 blocks allergic inflammations in the patients with asthma. Detection of interactions between gene and environment or between genes for complex disease such as asthma is important. We searched for an evidence of genetic effect of single nucleotide polymorphisms (SNPs) of MCP genes as well as gene - gene interactions involved in asthma. METHODS: Four hundreds asthmatics and four hundreds normal controls were enrolled. Asthma was defined as a positive bronchodilator response or positive methacholine provocation test with compatible clinical symptoms. Seven MCP gene SNPs (2 SNPs in MCP-1, 1 in MCP-2, and 4 in MCP-3) were included. Association analyses between SNP and asthma, and the tests for gene - gene interaction were performed. RESULTS: Strong linkage disequilibria were found among 7 MCP gene polymorphisms. There was no SNP that showed a significant association with asthma among 7 SNPs of 3 MCP genes. No haplotype was associated with asthma, either. The combination of MCP1-2518G>A, MCP2+46A>C, and MCP3+563C>T was the best predictive model for asthma as compared to the control in tests for gene - gene interaction. The MCP1-2518G>A and MCP2+46A>C was the second best predictive combination and this had the highest synergistic interaction effect on the subject's status than any other combination of polymorphisms. Complete linkages were not associated with the gene - gene interactions models. CONCLUSIONS: MCP gene polymorphisms probably interact with each other; thus, these findings may help in developing a possible genetic marker to predict asthma.

Gene - Gene Interactions Among MCP Genes Polymorphisms in Asthma

PURPOSE: Monocyte chemoattractant proteins (MCPs) are important cytokines that involved in cellular activation and releasing of inflammatoy mediators by basophils and eosinophils in allergic disease. Some MCP gene variants implicate in asthma and monoclonal antibody for MCP-3 blocks allergic inflammations in the patients with asthma. Detection of interactions between gene and environment or between genes for complex disease such as asthma is important. We searched for an evidence of genetic effect of single nucleotide polymorphisms (SNPs) of MCP genes as well as gene - gene interactions involved in asthma. METHODS: Four hundreds asthmatics and four hundreds normal controls were enrolled. Asthma was defined as a positive bronchodilator response or positive methacholine provocation test with compatible clinical symptoms. Seven MCP gene SNPs (2 SNPs in MCP-1, 1 in MCP-2, and 4 in MCP-3) were included. Association analyses between SNP and asthma, and the tests for gene - gene interaction were performed. RESULTS: Strong linkage disequilibria were found among 7 MCP gene polymorphisms. There was no SNP that showed a significant association with asthma among 7 SNPs of 3 MCP genes. No haplotype was associated with asthma, either. The combination of MCP1-2518G>A, MCP2+46A>C, and MCP3+563C>T was the best predictive model for asthma as compared to the control in tests for gene - gene interaction. The MCP1-2518G>A and MCP2+46A>C was the second best predictive combination and this had the highest synergistic interaction effect on the subject's status than any other combination of polymorphisms. Complete linkages were not associated with the gene - gene interactions models. CONCLUSIONS: MCP gene polymorphisms probably interact with each other; thus, these findings may help in developing a possible genetic marker to predict asthma.

Circulating MCP-1 level and tilde 2518 gene polymorphism as a marker of nephropathy development in Egyptian patients

Monocyte chemoattractant protein-1 [MCP-1] is a member of CC chemokine that plays an important role in the recruitment of monocytes/macrophages into renal tubulointerstitium. A biallelic A/G polymorphism at position tilde 2518 in the MCP-1 gene was found to regulate MCP-1 expression. MCP-1 and its A/G gene polymorphism have been implicated in the pathogenesis of some renal diseases. The aim of this study was to evaluate the role of circulating MCP-1 level and the relevance of functional genetic variations of MCP-1 as early predictors of the development of glomerulonephropathy [GN] in Egyptian patients. This is a case control study that was conducted in 50 GN patients, 20 non-GN cases and 20 ethnically matched healthy controls. MCP-1 serum level was detected by ELISA technique, while genotyping of polymorphisms in the MCP-1 genes was performed using a polymerase chain reaction [PCR] followed by restriction fragment length polymorphism [RFLP] detection High MCP-1 circulating levels and subsequently MCP-1tilde 2518G polymorphism are associated with the developing of nephropathy irrespective to the underlying etiology. MCP-1 serum level was significantly high when compared with healthy controls [P = 0.0007] and non-GN cases [P = 0.01]. There was predominance of A allele at tilde 2518 of MCP-1 gene in healthy controls [87.5%] and non-GN cases [77.5%]. The frequency of the tilde 2518G MCP-1 polymorphism was significantly higher in GN patients than in healthy controls [P <0.0001; OR= 15.6] and non-GN cases [P < 0.0001; OR = 7.7]. Interestingly, homozygosity for G allele plays the main role in such association. A/G polymorphism in MCP-1 gene and subsequently high circulating MCP-1 level confer a relevant role in the susceptibility to the development of nephropathy in the Egyptian population denoting that MCP-1 system could be an early predictor of such renal complication

Monocyte chemotactic protein-4 [MCP-4/CCL-13] and CC chemokine receptor 3 [CCR3] in the sputum of asthmatic children

Monocyte chemotactic protein-4 [MCP-4/CCL-13] is a potent chemoattractant to eosinophils, monocytes and lymphocytes. We aimed to investigate MCP-4 and its CC chemokine receptor 3 [CCR3] expression on cells of induced sputum during acute asthma exacerbation. Immunohistochemistry was used to assess MCP-4 and CCR3 expression on induced sputum cells of 30 children during asthma exacerbation and 20 healthy matched controls. Patients were divided into three groups according to exacerbation severity; mild, moderate and severe [n = 10 for each]. Patients were followed until quiescence, when sputum was re-examined. MCP-4 and CCR3 were expressed on eosinophils and monocytes. Lymphocytes expressed only MCP-4. The percentages of sputum total cells, eosinophils and lymphocytes expressing MCP-4 and/or CCR3 were significantly higher during asthma exacerbation than in controls and negatively correlated with peak expiratory flow rate, whereas that of monocytes was not. The percentages of sputum total cells, eosinophils, monocytes and lymphocytes expressing MCP-4; and total cells and eosinophils expressing CCR3 were significantly higher in patients with severe than those with mild and moderate exacerbations. When patients were followed till remission, the percentages of sputum cells expressing MCP-4 and CCR3 decreased. Sputum eosinophil percentage correlated positively with the percentage of eosinophils expressing MCP-4 and CCR3 [r = 0.69, p < 0.0001; r = 0.62, p < 0.001, respectively]. The percentage of sputum eosinophils expressing MCP-4 correlated positively with that of cells expressing CCR3 [r = 0.95, p < 0.0001]. The expression of MCP-4 and CCR3 on sputum cells increases during acute asthma exacerbation and this increase correlates with exacerbation severity, and it decreases during remission. Modification of their expression could be a potential target for asthma therapy

Alteração nos níveis de metaloproteinases da matriz e quimiocinas no fluido gengival durante o movimento dentário ortodôntico / Levels of the matrix metalloproteinases and chemokines in the gingival crevicular fluid of teeth under orthodontic forces

O objetivo do presente estudo foi avaliar os níveis das metaloproteinases da matriz MMP-9, MMP-3 e MMP-13, e das quimiocinas MCP-1, MIP-1β e RANTES, no fluido gengival de dentes sob força ortodôntica. Foram recrutados 14 pacientes (3 homens e 11 mulheres) que foram submetidos à movimentação ortodôntica dos caninos superiores. Amostras do fluido gengival foram coletadas com tiras de Periopaper em diferentes tempos. O volume do FG foi determinado com o uso do Periotron® e os niveis das MMPs e das quimiocinas quantificados usando-se uma multianálise imunoenzimática com microesferas. Os resultados mostraram que existe um aumento significativo no volume do FG na área de pressão em quase todos os tempos analisados, quando comparados às áreas de tensão. Os níveis da MMP-9 foram muito superiores aos das MMP-13 e MMP-3. Na análise da evolução do tempo pode ser observada uma alteração significativa nos níveis da quantidade total de MMP-9, MMP-13 e MMP-3 e na concentração de MMP-13 e MMP-3 durante o período de movimentação dentária no lado de pressão. A elevação dos níveis de expressão destas MMPs 1 hora após a aplicação da força ortodôntica sugere que estas enzimas estejam envolvidas na remodelação periodontal induzida. As quimiocinas MCP-1, MIP-1β e RANTES foram detectadas no sulco gengival em todos os diferentes intervalos de tempo analisados, nas áreas de tensão e de pressão. Porém, diversas amostras estavam abaixo do nível de detecção do ensaio e, os níveis dessas quimiocinas no fluido gengival não parecem ser alterados pelas forças ortodônticas.

The goal of the present study was to evaluate the levels of the matrix metalloproteinases MMP-9, MMP-3 and MMP-13 and of the chemokines MCP-1, MIP-1β and RANTES in the gingival crevicular fluid of teeth under orthodontic forces. Fourteen subjects (3 males and 11 females) were enrolled and subjected to orthodontic tooth movement of their maxillary canines. Samples of gingival crevicular fluid were collected from both tension and pressure sides using Periopaper strips at different time points. The volume of GCF was determined using a Periotron® and the levels of MMPs and chemokines quantified using a multiplex microbead immunoassay. The results demonstrated that the levels of MMP-9 were higher than the levels of MMP-13 and MMP-3. Statistically significant fluctuations during the orthodontic tooth movement could be detected for the total amount of MMP-9, MMP-13 and MMP-3 and for the concentration of MMP-13 and MMP-3 at the pressure side. The elevation in the levels of these MMPs, 1 hour after the application of the orthodontic force, suggests that these enzymes are involved in the induced periodontal remodeling. The chemokines MCP-1, MIP-1β and RANTES were detected in the GCF in both tension and pressure sides at different time points. However, several samples were below the level of detection of the assay and the levels of theses mediators in GCF did not seem to be altered by the orthodontic forces.

Activated natural killer cell-mediated immunity is required for the inhibition of tumor metastasis by dendritic cell vaccination

Immunization with dendritic cells (DCs) pulsed with tumor antigen can activate tumor-specific cytotoxic T lymphocytes (CTL), which is responsible for tumor protection and regression. In this study, we examined whether DCs pulsed with necrotic tumor lysates can efficiently prevent malignant melanoma tumor cell metastasis to the lung. DCs derived from mouse bone marrow were found to produce remarkably elevated levels of IL-12 after being pulsed with the tumor lysates. Moreover, immunization with these DCs induced CTL activation and protected mice from metastasis development by intravenously inoculated tumor cells. In addition, these DCs activated NK cells in vitro in a contact-dependent manner, and induced NK activities in vivo. Furthermore, NK cell depletion before DC vaccination significantly reduced the tumor-specific CTL activity, IFN-g production, and IFN-gamma- inducible gene expression, and eventually interfered with the antitumor effect of tumor-pulsed DCs. Finally, similar findings with respect to NK cell dependency were obtained in the C57BL/ 6J-bg/bg mice, which have severe deficiency in cytolytic activity of NK cells. These data suggest that the antitumor effect elicited by DC vaccination, at least in a B16 melanoma model, requires the participation of both cytolytic NK and CD8+ T cells. The findings of this study would provide important data for the effective design of DC vaccines for cancer immunotherapy.

Trypanosoma cruzi infection: a continuous invader-host cell cross talk with participation of extracellular matrix and adhesion and chemoattractant molecules

Several lines of evidence have shown that Trypanosoma cruzi interacts with host extracellular matrix (ECM) components producing breakdown products that play an important role in parasite mobilization and infectivity. Parasite-released antigens also modulate ECM expression that could participate in cell-cell and/or cell-parasite interactions. Increased expression of ECM components has been described in the cardiac tissue of chronic chagasic patients and diverse target tissues including heart, thymus, central nervous system and skeletal muscle of experimentally T. cruzi-infected mice. ECM components may adsorb parasite antigens and cytokines that could contribute to the establishment and perpetuation of inflammation. Furthermore, T. cruzi-infected mammalian cells produce cytokines and chemokines that not only participate in the control of parasitism but also contribute to the establishment of chronic inflammatory lesions in several target tissues and most frequently lead to severe myocarditis. T. cruzi-driven cytokines and chemokines may also modulate VCAM-1 and ICAM-1 adhesion molecules on endothelial cells of target tissues and play a key role in cell recruitment, especially of activated VLA-4+LFA-1+CD8+ T lymphocytes, resulting in a predominance of this cell population in the inflamed heart, central nervous system and skeletal muscle. The VLA-4+-invading cells are surrounded by a fine network of fibronectin that could contribute to cell anchorage, activation and effector functions. Since persistent "danger signals" triggered by the parasite and its antigens are required for the establishment of inflammation and ECM alterations, therapeutic interventions that control parasitism and selectively modulate cell migration improve ECM abnormalities, paving the way for the development of new therapeutic strategies improving the prognosis of T. cruzi-infected individuals

The experimental study on idiotypic DNA vaccine against human B-cell lymphoma to induce antitumor immune response / 中国实验血液学杂志

The purpose of this study was to evaluate whether the DNA vaccine containing idiotypic gene fragment of human B-cell lymphoma cell line Namalwa could elicit the specific anti-idiotypic immune response in vivo. The candidate gene fragment of the lymphoma cell, variable region of heavy chain (VH) of the membranous immunoglobulin, was amplified using Ig superfamily primers by means of RT-PCR. Also, the intact cDNA of murine monocyte chemoattractant protein (MCP-3) was cloned and used as the adjuvant molecular. The two gene fragments of VH and MCP-3 were fused together by 8aa linker peptide with recombinant PCR. Subsequently, the fusion gene fragment was cloned into eukaryonic expression vector pcDNA3.1 to construct DNA vaccine plasmid. Prior to the immunization, the transient transfection coupled with RT-PCR was performed to prove that the recombinant plasmid could express in eukaryonic cells in right way. Then two groups of mice were immunized by intramuscular injection with DNA vaccine and mock plasmid pcDNA3.1 respectively. Three times of injection were performed with 100 micro g plasmid respectively at the beginning of the experiment and 2, 4 weeks after the first injection for all the groups. FACS analysis was chosen to detect the antibodies recognizing lymphoma cells at different time following vaccination. The results demonstrated that specific anti-idiotypic antibody could be detected in the group of DNA vaccine immunized mice as early as eight weeks after the first immunization. Further test demonstrated that the anti-idiotypic antibody could maintain for at least twenty weeks with high titer. Anti-idiotypic antibodies were elicited in three of five mice of the DNA vaccine immunized group. The Abs of DNA vaccine immunized mice could only recognize Namalwa cell line instead of another unrelated human cell line A549. There is no cellular response detected in the DNA vaccine immunized mice. It is concluded that the DNA vaccine containing fused MCP3-VH sequence could elicit specific anti-idiotypic antibody against B-cell lymphoma in vivo and could be used in further study of DNA vaccine against B-cell lymphoma. The results would provide the basis for further studies and optimization of this therapeutic strategy on patients with B-lymphoproliferative disease.

Description of an in vivo model for assessment of eosinophil chemoattractants in the mouse

Chemokines (chemoattractant cytokines) induce potent and selective chemotaxis of leukocyte subsets in vitro. Here, we review briefly the chemokines shown to induce eosinophil chemotaxis in vitro and describe a novel model for the study of the ability of chemokines to stimulate eosinophil migration in vivo. Eosinophils were purified from the blood of mice over-expressing the IL-5 gene and labelled with 111In. Only the C-C chemokines, eotaxin and MIP-1 alpha, but not RANTES, MCP-1, MCP-3, MIP-1ß. KC and MIP-2, effectively induced the recruitment of 111In-eosinophils in mouse skin. We suggest that this mouse model will be useful in assessing the role of endogenously-generated chemokines in mediating eosinophil migration to sites of allergic inflammation in vivo.