Artesunate promotes Th1 differentiation from CD4+ T cells to enhance cell apoptosis in ovarian cancer via miR-142

The aim of this study was to evaluate the influence of artesunate on Th1 differentiation and its anti-tumor effect on ovarian cancer. A Murine ovarian cancer model was established by ID8 cells transplantation. The expression of miR-142 and Sirt1 proteins in peripheral CD4+ T cells were quantified with qRT-PCR and western blot, respectively. Peripheral CD4+ T cells were induced for Th1 differentiation. The percentages of apoptosis of Th1/CD4+ T cells and ovarian cancer cells were analyzed by flow cytometry. The IFN-γ level was examined through enzyme-linked immunosorbent assay. Artesunate promoted miR-142 expression in peripheral CD4+ T cells and Th1 differentiation from CD4+ T cells. Artesunate promoted cell apoptosis of ovarian cancer cells by inducing Th1 differentiation. By up-regulating miR-142, artesunate suppressed Sirt1 level and promoted Th1 differentiation. Artesunate enhanced the pro-apoptotic effects of Th1 cells on ovarian cancer via the miR-142/Sirt1 pathway. Artesunate promoted Th1 differentiation from CD4+ T cells by down-regulating Sirt1 through miR-142, thereby enhancing cell apoptosis in ovarian cancer.

Atorvastatin-medicated dentifrice significantly inhibits CD4+ T cell proliferation: in vitro pilot study / Pasta dental medicada con atorvastatina inhibe de forma significativa la proliferación de células T CD4+: un estudio piloto in vitro

Reports indicate that statins (cholesterol-lowering drugs), in addition to lowering cholesterol, have an immunomodulatory effect. This effect may be beneficial for the treatment of several diseases, including periodontal disease. The aim of the present study was to evaluate the immunomodulatory effect of an atorvastatin-medicated dentifrice on CD4+ T cell proliferation. CD4+ T cell proliferation assays and peripheral blood mononuclear cell (PBMC) viability assays were conducted on PBMCs from healthy donors cultured under the following conditions: control, atorvastatin solution, atorvastatin-medicated dentifrice, and dentifrice without atorvastatin at concentrations of 1, 5, 10, 50 and 100 µM. A Generalized Equation Estimation (GEE) model was used to analyze concentration versus proliferation and concentration versus percentage of dead cells within each group evaluated. Atorvastatin-medicated dentifrice (p-value <0.0001) and atorvastatin solution (p-value <0.0001) significantly inhibited CD4+ T cell proliferation in a dose-dependent manner compared with the dentifrice without atorvastatin and control conditions. Only the relationship between atorvastatin solution and percentage of dead cells was significant compared to the other conditions (p-value 0.019). The results revealed that atorvastatin-medicated dentifrice at concentrations of 1 to 100 µM had immunomodulatory effects, inhibiting CD4+ T cell proliferation without affecting PBMC viability. The other components of the dentifrice did not affect CD4+ T cell proliferation or cell viability, indicating its utility as a vehicle to achieve the desired effects of atorvastatin in periodontal tissue. Controlled clinical trials are still needed to evaluate the clinical effects of an atorvastatin-medicated dentifrice on the periodontium.

La literatura indica que las estatinas (medicamentos para bajar el colesterol), además de reducir el colesterol, tienen un efecto inmunomodulador. Este efecto puede ser beneficioso para el tratamiento de varias enfermedades, incluyendo la enfermedad periodontal. El objetivo de este estudio es evaluar el efecto inmunomodulador de una pasta dental medicada con atorvastatina sobre la proliferación celular de linfocitos T CD4+. A partir de células mononucleares de sangre periférica de donantes sanos (PBMC), se realizaron ensayos de proliferación y viabilidad de linfocitos T CD4+ bajo las siguientes condiciones: control, solución de atorvastatina, dentífrico medicado con atorvastatina y dentífrico sin atorvastatina, en concentraciones 1, 5, 10, 50 and 100 µM. Se realizó el análisis estadístico utilizando el modelo Generalized Equation Estimation (GEE) a fin de analizar la concentración versus la proliferación y la concentración versus el porcentaje de muerte celular para cada uno de los grupos. El dentífrico medicado con atorvastatina (valor p <0,0001) y solución de atorvastatina (valor p <0,0001) inhibieron significativamente la proliferación de células T CD4 + de una manera dependiente de la dosis en comparación con el dentífrico sin atorvastatina y condiciones de control. Sólo la relación entre la atorvastatina solución y el porcentaje de células muertas fue significativa en comparación con las otras condiciones (vale-p 0,019). Los resultados revelaron que el dentífrico medicado con atorvastatina en concentraciones de 1 a 100 mM tenía efectos inmunomoduladores, inhibiendo la proliferación de células T CD4 + sin afectar la viabilidad de PBMC. Los otros componentes del dentífrico no afectaron la proliferación de células T CD4 + o la viabilidad celular, indicando su utilidad como vehículo para conseguir los efectos deseados de atorvastatina en el tejido periodontal. Todavía se necesitan ensayos clínicos controlados para evaluar los efectos clínicos de un dentífrico medicado con atorvastatina sobre el periodonto.

Classical and alternative macrophages have impaired function during acute and chronic HIV-1 infection

Abstract Objectives: Three decades after HIV recognition and its association with AIDS development, many advances have emerged – especially related to prevention and treatment. Undoubtedly, the development of Highly Active Antiretroviral Therapy (HAART) dramatically changed the future of the syndrome that we know today. In the present study, we evaluate the impact of Highly Active Antiretroviral Therapy on macrophage function and its relevance to HIV pathogenesis. Methods: PBMCs were isolated from blood samples and monocytes (CD14+ cells) were purified. Monocyte-Derived Macrophages (MDMs) were activated on classical (MGM-CSF+IFN-γ) or alternative (MIL-4+IL13) patterns using human recombinant cytokines for six days. After this period, Monocyte-Derived Macrophages were stimulated with TLR2/Dectin-1 or TLR4 agonists and we evaluated the influence of HIV-1 infection and Highly Active Antiretroviral Therapy on the release of cytokines/chemokines by macrophages. Results: The data were obtained using Monocyte-Derived Macrophages derived from HIV naïve or from patients on regular Highly Active Antiretroviral Therapy. Classically Monocyte-Derived Macrophages obtained from HIV-1 infected patients on Highly Active Antiretroviral Therapy released higher levels of IL-6 and IL-12 even without PAMPs stimuli when compared to control group. On the other hand, alternative Monocyte-Derived Macrophages derived from HIV-1 infected patients on Highly Active Antiretroviral Therapy released lower levels of IL-6, IL-10, TNF-α, IP-10 and RANTES after LPS stimuli when compared to control group. Furthermore, healthy individuals have a complex network of cytokines/chemokines released by Monocyte-Derived Macrophages after PAMP stimuli, which was deeply affected in MDMs obtained from naïve HIV-1 infected patients and only partially restored in MDMs derived from HIV-1 infected patients even on regular Highly Active Antiretroviral Therapy. Conclusion: Our therapy protocols were not effective in restoring the functional alterations induced by HIV, especially those found on macrophages. These findings indicate that we still need to develop new approaches and improve the current therapy protocols, focusing on the reestablishment of cellular functions and prevention/treatment of opportunistic infections.

BJ-1108, a 6-Amino-2,4,5-trimethylpyridin-3-ol analogue, regulates differentiation of Th1 and Th17 cells to ameliorate experimental autoimmune encephalomyelitis

BACKGROUND: CD4+ T cells play an important role in the initiation of an immune response by providing help to other cells. Among the helper T subsets, interferon-γ (IFN-γ)-secreting T helper 1 (Th1) and IL-17-secreting T helper 17 (Th17) cells are indispensable for clearance of intracellular as well as extracellular pathogens. However, Th1 and Th17 cells are also associated with pathogenesis and contribute to the progression of multiple inflammatory conditions and autoimmune diseases. RESULTS: In the current study, we found that BJ-1108, a 6-aminopyridin-3-ol analogue, significantly inhibited Th1 and Th17 differentiation in vitro in a concentration-dependent manner, with no effect on proliferation or apoptosis of activated T cells. Moreover, BJ-1108 inhibited differentiation of Th1 and Th17 cells in ovalbumin (OVA)-specific OT II mice. A complete Freund's adjuvant (CFA)/OVA-induced inflammatory model revealed that BJ-1108 can reduce generation of proinflammatory Th1 and Th17 cells. Furthermore, in vivo studies showed that BJ-1108 delayed onset of disease and suppressed experimental autoimmune encephalomyelitis (EAE) disease progression by inhibiting differentiation of Th1 and Th17 cells. CONCLUSIONS: BJ-1108 treatment ameliorates inflammation and EAE by inhibiting Th1 and Th17 cells differentiation. Our findings suggest that BJ-1108 is a promising novel therapeutic agent for the treatment of inflammation and autoimmune disease.

Linfocitos TCD4+ y carga viral en pacientes con debut de sida que reciben tratamiento antirretroviral / CD4 + T lymphocytes and viral load in patients with AIDS debut receiving antiretroviral therapy

INTRODUCCIÓN: el debut de sida es una forma de presentación de la enfermedad causada por VIH que se caracteriza por alteración del estado general del paciente, síndrome de desgaste, aparición de graves infecciones oportunistas, neoplasias y alteraciones neurológicas. MÉTODOS: se estudió el comportamiento de los niveles de linfocitos T CD4+ y de carga viral en pacientes con debut de sida y terapia antirretroviral, al inicio y un año después del tratamiento. Se estudiaron 55 pacientes los cuales tuvieron al inicio del tratamiento conteos de linfocitos TCD4+ inferiores a 200cel/µL y carga viral elevada. RESULTADOS: después de un año de terapia, los valores de linfocitos T CD4+ aumentaron por encima de 200 cel/µL y la carga viral disminuyó a niveles no detectables en los pacientes estudiados. CONCLUSIONES: los resultados de esta investigación confirman los beneficios del tratamiento antirretroviral particularmente para los pacientes con debut de sida.

INTRODUCTION: the AIDS premiere is a form of presentation of the illness caused by HIV that is characterized by alteration of the patient's general state, waste syndrome, appearance of serious opportunists infections, neoplasia and neurological alterations. METHODS: the behavior of the CD4+ T lymphocytes levels were studied and of viral load in patient with AIDS premiere and antiretroviral therapy, to the beginning and one year after the treatment. 55 patients those were studied which had to the beginning of the treatment CD4+ T lymphocytes counts less to 200cel/µL and high viral load. RESULTS: after a year of therapy, the values of CD4+ T cells recovered and the viral load diminished at non detecting levels in the evaluated patients. CONCLUSIONS: the results of this study confirm the benefits of antiretroviral therapy, particularly for patient with AIDS premiere.

Attenuation of airway inflammation by simvastatin and the implications for asthma treatment: is the jury still out?

Although some studies have explained the immunomodulatory effects of statins, the exact mechanisms and the therapeutic significance of these molecules remain to be elucidated. This study not only evaluated the therapeutic potential and inhibitory mechanism of simvastatin in an ovalbumin (OVA)-specific asthma model in mice but also sought to clarify the future directions indicated by previous studies through a thorough review of the literature. BALB/c mice were sensitized to OVA and then administered three OVA challenges. On each challenge day, 40 mg kg-1 simvastatin was injected before the challenge. The airway responsiveness, inflammatory cell composition, and cytokine levels in bronchoalveolar lavage (BAL) fluid were assessed after the final challenge, and the T cell composition and adhesion molecule expression in lung homogenates were determined. The administration of simvastatin decreased the airway responsiveness, the number of airway inflammatory cells, and the interleukin (IL)-4, IL-5 and IL-13 concentrations in BAL fluid compared with vehicle-treated mice (P<0.05). Histologically, the number of inflammatory cells and mucus-containing goblet cells in lung tissues also decreased in the simvastatin-treated mice. Flow cytometry showed that simvastatin treatment significantly reduced the percentage of pulmonary CD4+ cells and the CD4+/CD8+ T-cell ratio (P<0.05). Simvastatin treatment also decreased the expression of the vascular cell adhesion molecule 1 and intercellular adhesion molecule 1 proteins, as measured in homogenized lung tissues (P<0.05) and human epithelial cells. The reduction in the T cell influx as a result of the decreased expression of cell adhesion molecules is one of the mechanisms by which simvastatin attenuates airway responsiveness and allergic inflammation. Rigorous review of the literature together with our findings suggested that simvastatin should be further developed as a potential therapeutic strategy for allergic asthma.

In vitro effects of meloxicam on the number, Foxp3 expression, production of selected cytokines, and apoptosis of bovine CD25+CD4+ and CD25-CD4+ cells

The purpose of this study was to evaluate the effect of meloxicam (MEL) on selected immune parameters of bovine CD25highCD4+, CD25lowCD4+, and CD25-CD4+ cells. Peripheral blood mononuclear cells (PBMCs) collected from 12-month-old heifers were treated with MEL at a concentration corresponding to the serum level of this medication following administration at the recommended dose (MEL 5 x 10(-6) M) and at a concentration 10 times lower (MEL 5 x 10(-7) M). After 12 and 24 h of incubation with the drug, the percentage of CD25highCD4+ cells decreased; however, this disturbance was quickly reversed. Furthermore, the absolute number of CD25highCD4+ cells in the PBMC populations treated with MEL 5 x 10(-6) M for 48 and 168 h was increased. Prolonged (168 h) exposure to the drug increased the percentage of Foxp3+ cells in the CD25highCD4+ cell subpopulation. The higher dose of MEL was found to significantly increase the percentage of IFN-gamma+ cells among the CD25-CD4+ cells. These results indicated that MEL does not exert an immunosuppressive effect by depleting CD4+ cells and suppression of IFN-gamma+ production by these cells. Furthermore, IL-10 and TGF-beta production was not changed following exposure to MEL.

Human Leukocyte Antigen-G Expression on Dendritic Cells Induced by Transforming Growth Factor-beta1 and CD4[+] T Cells Proliferation

During antigen capture and processing, mature dendritic cells [DC] express large amounts of peptide-MHC complexes and accessory molecules on their surface. DC are antigen-presenting cells that have an important role in tolerance and autoimmunity. The transforming growth factor-beta 1 [TGF-beta1] cytokine has a regulatory role on the immune and non-immune cells. The aim of this study is to evaluate the effect of TGF-beta1 on the induction of human leukocyte antigen-G [HLA-G] expression on the DC which is derived from monocyte. Methods: In this study, we evaluated the effect of TGF-beta1 in induction HLA-G expression on the monocyte-derived DC by flowcytometry and then CD4[+] T cell proliferative responses in the presence of DC-treated TGF-[beta1] was studied. Results: The results of this study showed that DC bearing HLA-G down-regulated activation of CD4[+] T cells and production of IL-6 and IL-17 in comparison with control [P<0.05]. Conclusion: It is concluded that TGF-beta1 has an important regulatory role in CD4[+] T cell proliferation by increasing HLA-G on DC and these cells can probably prevent unexpected immune responses in vivo

Prophylactic effect of Bovine lactoferrin against acute toxoplasmosis in immunocompetent and immunosuppressed mice

This study evaluated the immune-potentiating effect of administrating bovine Lactoferrin [LF] to immunocompetent [IC] and immunosuppressed [IS] mice prior to infection with tachyzoites of T. gondii RH strain. Mice were IS with cyclophosphamide. LF was given in seven of them as oral doses on alternate days. Immunological and parasitological assessments showed that LF induced statistical significance comparable resistance against acute toxoplasmosis in IC and IS mice. This was verified by elevated splenic CD4[+] T lymphocytes, reduced tachyzoites' viability and infectivity, with diminished parasite burdens. So, mice mortality declined and their survival was prolonged. This indicated that LF have prophylactic efficacy against human toxoplasmosis in risky persons with alleviating immune balance

Urine levels of rifampicin & isoniazid in asymptomatic HIV-positive individuals.

BACKGROUND & OBJECTIVE: AIDS and its associated gastrointestinal complications may impair the absorption of anti-tuberculosis (TB) drugs. Impaired absorption of anti-TB drugs could lead to low drug exposure, which might contribute to acquired drug resistance and reduced effectiveness of anti-TB treatment. The aim of this study was to obtain information on the status of absorption of rifampicin (RMP) and isoniazid (INH) in asymptomatic HIV- positive individuals, who are less immunocompromised. The D-xylose absorption test was also carried out to assess the absorptive capacity of intestive. METHODS: The absorption of RMP, INH and D-xylose was studied in 15 asymptomatic HIV-positive individuals with CD4 cell counts>350 cells/mm3 and 16 healthy volunteers, after oral administration of single doses of RMP (450 mg), INH (300 mg) and D-xylose (5 g). Urine was collected up to 8 h after drug administration. Percentage dose of the drugs and their metabolites and D-xylose excreted in urine were calculated. RESULTS: A significant reduction in the urinary excretion of INH and D-xylose in HIV-positive persons compared to healthy volunteers was observed. The per cent dose of RMP and its metabolite, desacetyl RMP was also lower in HIV-positive persons compared to healthy volunteers, but this difference was not statistically significant. INTERPRETATION & CONCLUSION: Decreased urinary excretion of D-xylose and INH are suggestive of intestinal malabsorption in HIV-positive individuals. HIV infection could cause malabsorption of anti-TB drugs even at an early stage of the disease. The clinical implications of these findings need to be confirmed in larger studies.

Unique features of bovine lymphocytes exposed to a staphylococcal enterotoxin

We previously demonstrated that stimulation of bovine peripheral blood mononuclear cells (PBMCs) with staphylococcal enterotoxin C (SEC), led to an inversion of the CD4(+):CD8(+) T cell ratio and generation of an atypical CD8(+) T cell subpopulation expressing CD26. In the present study, we examined T cell apoptosis and proliferation profiles of PBMC subpopulations in cultures stimulated with SEC. Unlike when stimulated with concanavalin A, nucleic acid synthesis in bovine PBMC cultures stimulated with SEC was low during the first four days but increased greatly on day 5. In contrast, nucleic acid synthesis in human PBMC cultures stimulated with SEC increased continuously. To investigate the mechanism of delayed bovine T cell proliferation, various cell phenotypes were monitored. The inversion of the bovine CD4(+):CD8(+) T cell ratio in PBMC cultures stimulated by SEC was associated with higher proliferation and lower apoptosis of CD8(+) T cells compared to CD4(+) T cells. The mRNA levels for interleukin (IL)-4 and IL-13 were sustained over 4 days but IL-12 mRNA levels dropped to background on day 2. These data suggest that SEC induces a prolonged Th-2- biased microenvironment, and together with the inversion of the bovine CD4(+):CD8(+) T cell ratios in bovine PBMC cultures with SEC, may in part explain the inability of the mammary immune system to establish an effective response to Staphylococcus aureus infections.

Prevention of chronic graft-versus-host disease by stimulation with glucocorticoid-induced TNF receptor

GITR (glucocorticoid-induced TNF receptor) is a recently identified member of the TNF receptor superfamily. The receptor is preferentially expressed on CD4+CD25+ regulatory T cells and GITR signals break the suppressive activity of the subset. In this study, we wanted to reveal the in vivo function of GITR in chronic graft-versus-host disease (cGVHD), a lupus-like autoimmune disease. A single injection of anti-GITR monoclonal antibody (DTA-1) was effective in blocking the progression of cGVHD in the parent-into-F1 model. Treatment of DTA-1 significantly decreased levels of IgG1 anti-DNA autoantibody, inhibited glomerulonephritis, and increased survival. The DTA-1-mediated inhibition of autoantibody production correlated with deletion of B cells and could occur independently of CD4+CD25+ regulatory T cells. Our results indicate that anti-GITR monoclonal antibody may be used as a potential immunotherapeutic agent for preventing cGVHD.

Presentación de cinco casos de linfocitopenia CD4+ idiopática / Report of five cases of idiopathic CD4+ lymphocytopenia

Introducción: el objetivo de este trabajo es llamar la atención sobre una inmunodeficiencia recientemente descripta y ofrecer una propuesta de estudio y tratamiento. Material y métodos: período de estudio: enero de 1998 a agosto de 2000. Presentamos 5 pacientes de entre 23 y 86 años de edad (media 51,2 años), todos de sexo femenino, que fueron estudiadas por presentar infecciones a repetición (micóticas, virales y bacterianas). Se les realizó determinación de VIH por ELISA, pruebas cutáneas para antígenos habitualmente reconocidos por el organismo, medición de linfocitos CD4+ y CD8+ por partículas magnéticas. Proteinograma electroforético, dosaje de inmunoglobulinas plasmáticas por inmunodifusión radial. Se efectuó tratamiento con timomodulina 60 mg por día. Conclusiones: al momento del diagnóstico se observó: pruebas cutáneas hipoérgicas, VIH negativo y disminución de linfocitos CD4+. Posterior al tratamiento con timomodulina, se observó buena respuesta clínica, mejoría de las pruebas cutáneas y elevación de los linfocitos CD4+ en todas las pacientes

Impacto de la infección por el virus de la inmunodeficiencia humana Tipo-1 en la población hemofílica / The impact of human immunodeficiency virus infection in haemophilic patients

En los últimos 3 años se han producido notables avances en el tratamiento de la enfermedad HIV/SIDA. En este trabajo se compara la evolución y respuesta al tratamiento con antirretrovirales en 175 pacientes hemofílicos HIV (+) evaluados entre los años 1983 y 1995 con 54 pacientes de las mismas características, pero que recibieron terapias combinadas de alta eficacia, entre los años 1996 y 1999. Se evaluaron los siguientes parámetros: influencia de la edad al momento de la infección con el tiempo de sobrevida libre de enfermedad, incidencia de complicaciones oportunistas, tipo y severidad de la hemofilia en relación con el riesgo de infección por el retrovirus, respuesta al tratamiento antirretroviral, efectos adversos relacionadas con el mismo y modificaciones acaecidas en los niveles de linfocitos T CD4 (+) y en los valores de la carga viral en plasma.

Effect of zinc supplementation on zinc status, copper status and cellular immunity in elderly patients with diabetes mellitus.

This preliminary study was to investigate the effects of zinc supplementation on Zn status, Cu status, serum macrominerals and lymphocyte subsets in elderly diabetic patients. The results of Zn supplementation can correct plasma Zn levels to normal values. However, this treatment did not affect the cellular Zn, copper status and serum macrominerals. Enhancement of the percentage of CD4 cells was observed after Zn therapy but had no effect on the percentage of CD8 cells and CD4/CD8 ratios. Our finding implicated that zinc supplementation might be useful to enhance the immune status in these patients.

Antiviral activity of interferon gamma in vivo during mouse hepatitis virus infection

A/J mice became resistant to experimental MHV3 infection after immunization with UV-inactivated MHV3 (0 percent mortality, 0/10). Depletion of interferon (IFN) gamma-producing CD4+ T lymphocytes with monoclonal antibodies to CD4+ led to susceptibility to virus infection (60 percent of mortality, 6/10). The resistance to MHV3 infection of CD4+ T lymphocyte-depleted-A/J mice was restored by treatment with 1000 U of IFN gamma on days -1, 0, 1, 2, 3 and 4 (10 percent of mortality, 1/10). The low virus titers observed in resistant mice (controls or CD4+ depleted plus IFN gamma treated) were cleared 6 days after infection and the virus titers observed among susceptible mice (CD4+ depleted) increased gradually and peaked on day 6, when the animals died. Previous data, taken together with the direct evidence presented in this paper, provide strong evidence supporting the concept of an in vivo antiviral role of IFN gamma through a central action on the mechanisms of resistance to MHV3 infection

Effect of high dose intravenous pulse dexamethasone on lymphocyte phenotypes.

The lymphocyte phenotypes were enumerated in 10 patients with collagen diseases at 0 h, 4 h, 24 h and 7 days after a megadose (100 mg) iv pulse dexamethasone. A significant decrease in CD3 (from a mean of 2324.3/mm3 to 705.9/mm3) and CD4 (from a mean of 1642.6 to 317.6/mm3) cells was observed at 4 h, which recovered partially by 24 h (186.7 and 1226.3/mm3 respectively) and completely at 7 days (2496.1 and 1838.4/mm3). A transient decrease in CD8 cells at 4 h was also observed. There was no significant effect on B cells.