Sodium Selenite Diminished the Regulatory T Cell Differentiation In Vitro.

Sodium selenite modulates the activity of lymphocytes. It negatively regulates the suppressive activity of cells and increases the immune response. In this study, we evaluated whether the regulatory T cell differentiation was modulated by sodium selenite. The percentages of CD4+CD25+Foxp3+, CD4+CD25+, and CD4+CTLA-4+ cells in CD4+ T cells cultures stimulated with IL-2 and TGF-ß in the presence or absence of selenium, in the form of sodium selenite (2.0×10-6M), were evaluated by flow cytometry. The mRNA expression of TET2/3 enzymes and IL-10 was analyzed by RT-qPCR and the levels of IL-10 were measured by an ELISA. We observed a decrease in CD4+CD25+Foxp3+ and CD4+CTLA-4+ cells in presence of selenium. However, normal percentages were reached again after selenium removal. An increase in CD4+CTL4-4+ cells was detected in selenium-primed cell cultures in absence of IL-2 and TGF-ß. In addition, we observed a decrease in TET3 in presence of selenium. Finally, we observed an augment in IL-10 transcription and protein levels and relative expression of TET2 in cultures exposed to selenium. We suggest that selenium reversibly affects the regulatory T cell differentiation in vitro. Likewise, selenium may modulate Treg percentages promoting optimal immune responses and, at the same time, the expression of specific suppressor molecules.

AIM2-inflammasome role in systemic lupus erythematous and rheumatoid arthritis.

The inflammasome AIM2 regulates multiple aspects of innate immune functions and serves as a critical mediator of inflammatory responses. AIM2 inflammasome activation leads to the production of pro-inflammatory cytokines, IL-1ß and IL-18 and participates triggering a pyroptosis response needed to counteract excessive cell proliferation. In addition, AIM2 expression and activation is wide regulated since alteration in its activity may derived in pathological consequences. Consequently, deregulated AIM2 activation contributes to the pathogenic processes of various inflammatory diseases. In this review, we will discuss the activation and function of AIM2 inflammasome, as well as its contribution in rheumatoid arthritis and systemic lupus erythematous pathology. Finally, we highlight the participation of the AIM2-inflammasome at the level of joint in rheumatoid arthritis and at kidney in systemic lupus erythematous. The development of therapeutic strategies based on modulation of AIM2-inflammasome activity should have a tissue-specific focus.

Expression of mBD4, mBD3 and CRAMP during type II collagen-induced arthritis/CIA and their association with inflammation and bone-remodeling markers.

The aim of this study was to analyze the expression of mBD4, mBD3 and CRAMP in joint of mice with type II collagen-induced arthritis/CIA and to explore its possible association with IL-10, IL-4, IFN-γ, IL-17, MMP3, RANK/RANKL/OPG and histological parameters. METHODS: CIA was induced in 44 DBA/1 J mice. The joints from mice were classified into the onset, peak and remission phase of CIA. Histological sections were stained with hematoxylin-eosin and safranin O. The expression of CRAMP, mBD-3, mBD-4, and MMP-3 was evaluated using reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The expression of IL-10, IL-4, IFN-γ, IL-17, RANK/RANKL/OPG was analyzed by RT-PCR. RESULTS: We observed that inflammation and immunostained cells for CRAMP increased in the peak and remission phases compared to the control group. In addition, increments in relative expressions of CRAMP were detected for the remission phase and in IL-4 and IL-17 in the peak phase compared to the control and onset phase. In addition, an increase in IL-10 in a peak phase compared to the control, as well as the relative expression of IFN-γ in remission phase was higher than in the onset phase. This was accompanied by an increase in cartilage damage in the peak phase compared to the control. Cells immunostained to MMP3 increased in the peak phase compared to the onset and control group, and relative expression of MMP3 was detected in the peak phase compared to the onset, remission, and control group. We observed that the relative expression of RANK and RANKL in the peak phase was higher than in control and onset phase. Finally, the relative expression of OPG in the peak phase compared to the onset, remission, and control group was detected. Regarding CRAMP behavior in the different phases studied, it was positively correlated with IL-4 and RANK, and showed a negative correlation with IFN-γ, IL-17, IL-10, RANKL, OPG and RANKL/OPG ratio in the control group. Also was positively correlated with IFN-γ, IL-17, IL-4, IL-10, as well as with RANK, RANKL, and OPG in the onset and peak phases of the CIA. In the peak phase, CRAMP showed a positive association with MMP3, and we observed a direct correlation between CRAMP and IFN-γ and RANKL/OPG ratio in remission phase. mBD3 correlates positively with IFN-γ, IL-17, IL-10, RANKL, OPG and RANKL/OPG ratio, and showed a negative correlation with CRAMP, MMP3, and RANK in the control group. Also, it was directly associated with IFN-γ, IL-17, IL-4, IL-10 and RANKL in the onset phase while it was inversely associated with CRAMP, MMP-3, RANK, RANKL, and OPG in the peak phase. Finally, mBD3 was inversely correlated with MMP3 in the remission phase and was directly associated with CRAMP, IFN-γ and RANKL/OPG ratio in this phase. mBD4 was directly associated with CRAMP, IFN-γ, IL-17, IL-4, IL-10, RANKL / OPG in the onset phase, and with CRAMP, IFN-γ, IL-17, IL-4, IL-10, MMP3, RANK, RANKL and OPG in the peak phase. Finally, mBD4 was positively associated with mBD3, IFN-γ, IL-17, IL-10, RANK, RANKL OPG and RANKL/OPG in the CIA remission phase. CONCLUSIONS: Our results demonstrate that CRAMP plays an important role in CIA progress and suggest that its abundance is associated with local pro- and anti-inflammatory status. This makes us propose CRAMP as a possible contributor of bone reconstruction in the last stage of CIA.

B regulatory cells associated with changes in biochemical and inflammatory parameters in normal-glycemic individuals, pre-diabetes and T2DM patients.

AIM: To determine the percentages of (CD19 + CD24 + CD38+, CD19 + CD24 + CD27+, CD19 + IL-10+)-Breg cells, IL-17 single and IL-17+/IFN-γ double producers T cells and IFN-γ+ T cells, in normal-glycemic individuals, prediabetes and T2DM patients, and to analyze the association of Breg cells with metabolic parameters of T2DM. METHODS: percentages of Breg cells, IL-17+ and IL-17 + IFN-γ+ T cells, IFN-γ+ T cells and IL-10 were determined by flow cytometry. IL-6 levels were evaluated by ELISA assay. RESULTS: increased IL-6 levels, IL-17+ and IL-17 + IFN-γ+ T cells and a diminution of IL-10 levels and CD19 + IL-10+ cells in T2DM patients were observed. We found that CD19 + CD24 + CD27+ cells and CD19 + CD24 + CD38+ cells were increased in T2DM patients. The percentages of CD19 + CD24 + CD38+ cells were associated with HOMA-B, TyG index, HDL and cholesterol values. In normal-glycemic individuals, CD19 + CD24 + CD27+ cells were inversely associated to triglycerides and TyG index. In prediabetes patients, CD19 + CD24 + CD38+ cells were inversely related with cholesterol and LDL. Finally, CD19 + CD24 + CD38+ cells were inversely related with HDL values in T2DM patients. CONCLUSION: Our results suggest that increased percentages of IL-17 single and IL-17/IFN-γ double producers T cells in T2DM patients may be a consequence of the initial CD19 + IL-10+ cells reduction. Furthermore, dyslipidemia could play an important role in percentages and activity of B regulatory cells.

The inflammatory state of adipose tissue is not affected by the anti-inflammatory response of the A2a-adenosine system and miR-221/PTEN.

Obesity is a chronic inflammatory state with cytokines, adipokines, and miRNAs. The A2a-adenosine system decreases activation and cytokine release in immune cells. MiR-221 is upregulated in carcinogenesis and inflammatory processes, where its targets PTEN and ETS-1, negatively regulates the Akt pathway and induces the release of pro-inflammatory cytokines, respectively. However, the roles of the A2a-adenosine system and miR-221 in adipose tissue are unknown. The aim of this work was to evaluate the A2a-adenosine and miRNA pathways as immune modulators in adipose tissue. We collected aspirate of adipose tissue from patients with BMI < 25 kg/m2 (BMI < 25) and BMI ≥ 25 kg/m2 (BMI ≥ 25) who underwent liposuction; the adipose tissue was digested with collagenase, and then a Ficoll gradient was performed to obtain mononuclear cells from adipose tissue (MCAT). We evaluated the A2a levels by quantitative Retro-transcriptase Polymerase Chain Reaction (RT-qPCR) and flow cytometry and the A2a-adenosine function with a proliferation assay or cytokine levels in the presence or absence of NAD+, activators, and inhibitors of the system. We also analyzed miR-221, ETS-1 and PTEN levels by qRT-PCR. First, we detected that MCAT presented higher basal proliferation than mononuclear cells from peripheral blood; however, activation of the A2a receptor downregulated cell proliferation and cytokine release. Interestingly, while miR-221 was downregulated in MCAT from subjects with BMI ≥ 25 compared to BMI < 25, their targets ETS-1 and PTEN, were increased. In conclusion, the A2a-adenosine system is decreased in MCAT, but it maintains its function; moreover, miR-221 could participate in promoting inflammation in adipose tissue.

Estatus funcional de adultos mayores de Tláhuac, Ciudad de México / Functional status of senior adults in Tlahuac, Mexico City / Status funcional de idosos de Tláhuac, Cidade do México

Resumen La identificación de la capacidad de funcionalidad en el adulto mayor es una valoración conveniente. Se concibe que la funcionalidad está relacionada con lograr satisfacer sus necesidades básicas, bienestar emocional y social, significa independencia y, por lo tanto, con calidad de vida. El objetivo de este estudio fue identificar el grado de funcionalidad que presentan los adultos mayores de 60 años y mayores que asisten a consulta en 4 centros de salud ubicados en la delegación Tláhuac, Ciudad de México durante el periodo de junio a diciembre de 2014. Fue una investigación exploratoria, en una muestra por conveniencia de 754 adultos mayores; se utilizaron 5 ítems del apartado A del instrumento "Evaluación Funcional del Adulto Mayor" el cual fue aplicado en los centros de salud. Para el procesamiento de la información se utilizó estadística descriptiva. Resultados: El 82% de los adultos mayores fueron independientes y el 18% están en riesgo de dependencia. Las actividades que presentaron menor porcentaje de dependencia fueron preparar sus alimentos y realizar tareas domésticas. Conclusión: Los resultados arrojados en el presente estudio implican sensibilizar a los profesionales de los centros de salud para que incorporen en sus programas propuestas para mantener la funcionalidad en este grupo etario como son ejercicio mental y físico, hábitos alimentarios, ocupación del tiempo y el manejo de situaciones estresantes.

Abstract It is important to address the functional status of senior adults because this is an indicator of their capacities to fulfill their basic needs, as well as to have emotional and social wellbeing, and thus to live a life with quality and independence. The objective of this study was to identify the degree of functionality in seniors 60 years and older who visit four health centers in Tlahuac, Mexico City. Data were collected from June to December 2014. This was an exploratory study on a convenience sample of 754 senior adults. Five items from the section A of the instrument Senior Adult Functional Status (Evaluación Funcional del Adulto Mayor (EFAM)) were used. The questions were asked in the health centers. Descriptive statistics were performed. Results: 82% of the seniors showed being independent while 18% were at risk of being dependent. The activities which demonstrated lower percentage of dependence were preparation of their food and performing their domestic tasks, Conclusion: The findings of this study suggest that health professionals should be sensitized to incorporate proposals in their programs to help maintain the functional status of this population. These activities should focus on: physical and mental exercise, good dietary habits, adequate use of time, and stress management.

Resumo A identificação da capacidade de funcionalidade em idosos é uma valorização conveniente. Concebendo que a funcionalidade está relacionada em conseguir satisfazer suas necessidades básicas, bem-estar emocional e social, significa independência, neste conceito a qualidade de vida está considerada. O objetivo deste estudo foi identificar o grau de funcionalidade que apresentam os idosos de 60 anos e mais velhos, que assistem a consulta em quatro centros de saúde localizados no município Tláhuac, Cidade de México, durante o período de junho a dezembro de 2014. Foi uma pesquisa exploratória, em uma amostra de conveniência de 754 idosos, utilizaram-se cinco itens do apartado A do instrumento "Avaliação Funcional do Idoso" (EFAM), o qual foi aplicado nos centros de saúde. Para o processamento da informação utilizou-se estatística descritiva. Resultados: O 82% dos idosos foram independentes e o 18% estão em risco de dependência. As atividades que apresentaram menor porcentagem de dependência foram preparar seus alimentos e realizar fazeres domésticos. Conclusão: Os resultados obtidos no presente estudo procuram sensibilizar aos professionais dos centros de saúde para que incorporem nos seus programas, propostas para manter a funcionalidade neste grupo etário como são o exercício físico e mental, hábitos alimentares, ocupação do tempo e o manejo de situações estressantes.

Regulatory T-cell subsets in response to specific Mycobacterium tuberculosis antigens in vitro distinguish among individuals with different QTF and TST reactivity.

Regulatory T cells (Tregs), a subset of CD4+ T cells related with immune regulation, have been associated with active and latent tuberculosis infection (LTBI). Treg frequencies were evaluated by multicolor flow cytometry (FC) in peripheral blood mononuclear cells (PBMCs) stimulated with mycobacterial antigens ESAT-6, CFP-10, and TB7.7 to assess their capacity to distinguish subjects with different reactivity to the QuantiFERON-TB® Gold In-Tube (QFT-IT) test and the tuberculin skin test (TST). Increased frequencies of CD4+CD25highCD39+ cells were found for the [TST+, QTF+] compared with the [TST+, QTF-] group. Also, higher frequencies were observed for the [TST+, QTF+] compared with the [TST+, QTF-] and [TST-, QTF-] groups in CD4+CD25highFoxp3+ and CD4+CD25highCD39+Foxp3+ populations. Receiver operating characteristics (ROC curve) analysis confirmed these discriminating results. QFT-IT and TST quantitative values correlated with several Treg population frequencies.

Immunomodulatory effect of diethylcarbamazine in mice infected with Nocardia brasiliensis.

We tested whether diethylcarbamazine (DEC) or ivermectin (IVM), both antiparasitic drugs with reported immunomodulatory properties, were able to affect the immune system to potentiate host defense mechanisms and protect against actinomycetoma in a mouse model. Male BALB/c mice of 10-12 weeks of age were injected with either Nocardia brasiliensis or saline solution. Recorded were the effects of a treatment by DEC (6 mg/kg per os daily for one week) or IVM (200 µg/kg subcutaneously on days 1 and 3) on (i) the development of mycetoma lesion, (ii) the expression of reactive oxygen intermediates (ROI) by phagocytes, (iii) the proliferation index of lymphocytes and (iv) antibody production of IgG and IgM. After an initial lesion in all mice, DEC inhibited a full development and progression of actinomycetoma resulting in a reduced lesion size (p < 0.001). IVM had no inhibitory effect on the development of mycetoma. Furthermore, DEC treatment was associated with a significant enhancement of ROI expression (p < 0.05) by polymorphonuclear neutrophils at day 3 after infection. Lymphocyte proliferation in response to N. brasiliensis antigens and concanavalin A in DEC-treated group was higher than in non-treated group at day 21 and 28 postinfection (p < 0.01). Significant changes in antibody response were not observed. By all parameters tested, DEC was superior to IVM regarding immunostimulatory potency. In conclusion, DEC expressed an in vivo influence on the immune status during the infection by N. brasiliensis leading to retrogression of the mycetoma and increasing cellular immune responses. Our findings may indicate a potential use of DEC as a putative adjuvant in infectious disease or vaccination.

Analysis of Th1, Th17 and regulatory T cells in tuberculosis case contacts.

We have hypothesized that individuals infected with Mycobacteriumtuberculosis that exhibit different patterns of immune reactivity in serial interferon (IFN)-γ release assays (IGRA's) correspond to different status within the immune spectrum of latent tuberculosis (TB). Accordingly, we analyzed the possible association between the consistent results (negative or positive) in an IGRA test and relevant immune parameters, mainly the levels of Th1 and Th17 lymphocytes and T regulatory (Treg) cells in the peripheral blood of TB case contacts. We found that individuals with a persistently positive IGRA showed increased levels of Th1 and Th17 lymphocytes upon in vitro stimulation with MTB antigens. In addition, a significant increase in the proportion of CD4+CTLA-4+ and CD4+Foxp3+ cells was detected in assays with blood samples from these individuals. Our data support that different immune phenotypes can be identified into the spectrum of latent TB, by combining different parameters of immune reactivity against MTB.

In vitro assessment of agave fructans (Agave salmiana) as prebiotics and immune system activators.

The prebiotic effect of agave fructans (Agave salmiana) was evaluated through the growth of two lactic acid bacterial (LAB) strains (Lactobacillus casei and Bifidobacterium lactis). The immune system was activated through the stimulation of peripheral blood mononuclear cells (PBMC) of healthy subjects testing fructans, LAB or a mixture of these compounds at different concentrations. Immune responses, such as early cell activation (CD69), cell cycle progression, nitric oxide (NO) production and the expression of transcription factors for lymphocyte differentiation, were analyzed. Compared with other fructans, the extracted agave fructans showed the highest prebiotic activity and increased levels of CD69 expression, proliferative activity and NO production when administered with the probiotic L. casei. The Th1 lymphocyte differentiation produced through LAB stimulation was greatly diminished after the incorporation of agave fructans. In conclusion, these types of fructans (A. salmiana) are involved in the activation and selective differentiation of cells of the immune system through interactions with probiotics. Thus, agave fructans represent a novel immunomodulator that might benefit the functional food industry.

Unsupervised cardiac image segmentation via multiswarm active contours with a shape prior.

This paper presents a new unsupervised image segmentation method based on particle swarm optimization and scaled active contours with shape prior. The proposed method uses particle swarm optimization over a polar coordinate system to perform the segmentation task, increasing the searching capability on medical images with respect to different interactive segmentation techniques. This method is used to segment the human heart and ventricular areas from datasets of computed tomography and magnetic resonance images, where the shape prior is acquired by cardiologists, and it is utilized as the initial active contour. Moreover, to assess the performance of the cardiac medical image segmentations obtained by the proposed method and by the interactive techniques regarding the regions delineated by experts, a set of validation metrics has been adopted. The experimental results are promising and suggest that the proposed method is capable of segmenting human heart and ventricular areas accurately, which can significantly help cardiologists in clinical decision support.

Quantitative analysis of regulatory T cells in kidney graft recipients: a relationship with calcineurin inhibitor level.

BACKGROUND: Monitoring of immunosuppressive drug levels can prevent some adverse effects in patients with solid organ transplantation. However, the possible relationship between these drug levels and their biological effect on immune cells has not been studied in depth. The aim of this work was to assess the possible effect of immunosuppressive therapy with calcineurin inhibitors on the levels of regulatory T cells in patients with kidney transplantation. METHODS: Serial samples of peripheral blood were obtained from six patients that underwent kidney transplantation and received a triple pharmacological therapy, which included a calcineurin inhibitor (Cyclosporine A or Tacrolimus). Levels of CD4+CD25(high), CD4+Foxp3+ and CD8+CD28- cells and calcineurin inhibitors were evaluated by flow cytometry, and by radioimmunoassay, respectively. In addition, we assessed the in vitro effect of cyclosporine and tacrolimus on the number of T regulatory cells in peripheral blood mononuclear cells from healthy subjects. RESULTS: Diminished levels of total CD8+ T cells were found at week 1 after kidney transplantation, with a recovery of this subpopulation at week twelve. In addition, a significant decrease in CD8+CD28- T cell levels was observed at weeks 4 and 12 after transplantation. In contrast, no significant differences were observed in the levels and function of CD4+CD25(high) or CD4+Foxp3+ regulatory T cells. Furthermore, no significant correlation between the level/dose ratios of calcineurin inhibitors and the percentages of regulatory cells was detected. On the other hand, in vitro assays showed that cyclosporine (5.0 ug/mL) and tacrolimus (100 ng/mL) diminished the percentages of CD8+CD28- cells, with no significant effect on natural T regulatory cells. CONCLUSIONS: Our results suggest that although calcineurin inhibitors do not have a significant effect on the level and function of CD4+CD25(high) or CD4+Foxp3+ regulatory T cells in patients with kidney transplantation, these drugs seem to exert a down-regulatory effect on CD8+CD28- lymphocytes.

Transposicion corregida de los grandes vasos. Tratamiento quirurgico de la comunicacion interventricular. / Corrected transposition of the great vessels. Surgical treatment of interventricular communication

Se informa de tres casos de transposicion corregida de los grandes vasos que acompana a la comunicacion interventricular, y que fueron enmendados por el ventriculo sistemico, con dos sobrevivientes. No se encontro evidencia de lesion en los tejidos de conduccion de los pacientes que sobrevivieron. Esta via de acceso ofrece la ventaja de poder cerrar la comunicacion interventricular con los tejidos de conduccion lejanos al borde por este ventriculo, especialmente cuando no se tiene un aparato identificador de haz de Hiss