RESUMEN
Transferon® is a complex drug based on a mixture of low molecular weight peptides. This biotherapeutic is employed as a coadjuvant in clinical trials of several diseases, including viral infections and allergies. Given that macrophages play key roles in pathogen recognition, phagocytosis, processing, and antigen presentation, we evaluated the effect of Transferon® on phenotype and function of macrophage-like cells derived from THP-1 monocytes. We determined the surface expression of CD80 and CD86 by flow cytometry and IL-1ß, TNF-α, and IL-6 levels by ELISA. Transferon® alone did not alter the steady state of PMA-differentiated macrophage-like THP-1 cells. On the contrary, simultaneous stimulation of cells with Transferon® and LPS elicited a significant increase in CD80 (P ≤ 0.001) and CD86 (P ≤ 0.001) expression, as well as in IL-6 production (P ≤ 0.05) compared to the LPS control. CD80 expression and IL-6 production exhibited a positive correlation (r = 0.6, P ≤ 0.05) in cells exposed to Transferon® and LPS. Our results suggest that the administration of Transferon® induces the expression of costimulatory molecules and the secretion of cytokines in LPS-activated macrophages. Further studies are necessary to determine the implication of these findings in the therapeutic properties of Transferon®.
Asunto(s)
Antígeno B7-1/genética , Interleucina-6/inmunología , Lipopolisacáridos/farmacología , Macrófagos/efectos de los fármacos , Macrófagos/inmunología , Factor de Transferencia/farmacología , Antígeno B7-1/inmunología , Antígeno B7-2/genética , Antígeno B7-2/inmunología , Diferenciación Celular/efectos de los fármacos , Citocinas/inmunología , Citometría de Flujo , Humanos , Recuento de Leucocitos , Monocitos/efectos de los fármacos , Células THP-1RESUMEN
Prostate cancer (PCa) is the second most frequently diagnosed cancer in men worldwide. Dialyzed Leukocyte Extracts (DLEs) are heterogeneous mixtures of low-molecular-weight peptides that improve clinical responses in various diseases. Here, we analyzed the effects of TransferonTM, a commercial DLE with characterized active pharmaceutical ingredient and proven batch-to-batch reproducibility, in preclinical models of PCa. We employed v-Src-transformed murine prostate epithelial (PEC-Src) cells, which recapitulate the transcriptional profiles in human PCa, can be grown in immunocompetent mice, and consistently form bone and brain metastases. In vitro, TransferonTM did not induce cytotoxicity nor alterations in migration /invasion of PEC-Src cells. In vivo, TransferonTM reduced metastatic dissemination after intracardiac injection of PEC-Src and inhibited tumor growth of subcutaneous isotransplants. The antineoplastic effect of TransferonTM correlated with changes in tumor infiltration, increased serum concentrations of IL-12 and CXCL1, and reduced levels of VEGF. Our results suggest that the antineoplastic effect produced by TransferonTM is due to its immunomodulatory activity and not by a direct effect on cancer cells, and indicate that TransferonTM could be beneficial as adjuvant therapy in PCa patients.
Asunto(s)
Neoplasias Encefálicas/secundario , Neoplasias de la Próstata/tratamiento farmacológico , Neoplasias de la Próstata/patología , Factor de Transferencia/uso terapéutico , Animales , Antineoplásicos/farmacología , Muerte Celular/efectos de los fármacos , Línea Celular Tumoral , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Modelos Animales de Enfermedad , Interleucina-1/metabolismo , Interleucina-12/metabolismo , Masculino , Ratones , Invasividad Neoplásica , Factor de Transferencia/farmacología , Factor A de Crecimiento Endotelial Vascular/metabolismoRESUMEN
OBJECTIVES: To evaluate the effect of dialysable leucocyte extract (DLE) on pro- and anti-inflammatory profiles in a rat model of osteoarthritis (OA). METHOD: Forty-eight male Wistar rats were divided into three groups: normal rats without treatment, OA rats treated with placebo, and OA rats treated with DLE. After treatment, the animals were killed to obtain cartilage for histological analysis and to determine the expression of pro- and anti-inflammatory cytokines by reverse transcription multiplex polymerase chain reaction (RT-MPCR) and immunohistofluorescence analyses. RESULTS: Histological analysis revealed that OA cartilage from rats treated with DLE displayed similar characteristics to non-OA cartilage from the control group. The OA cartilage treated with placebo showed alterations in the cellular architecture and in chondrocyte cluster formation. Analysis of cytokine expression by RT-MPCR showed that OA cartilage from DLE-treated rats expressed platelet-derived growth factor (PDGF), interferon (IFN)-γ, and fibroblast growth factor (FGF)-2, similar to non-OA cartilage from the control group. However, OA cartilage from rats treated with placebo expressed interleukin (IL)-1, PDGF, and I kappa B (IκB). Confocal immunodetection of FGF-2, PDGF, and non-phosphorylated IκB showed that they were distributed in the cytoplasm of most chondrocytes in OA cartilage from DLE-treated rats whereas no nuclear factor kappa B (NF-κB) expression was observed in the nuclei. Instead, in OA cartilage from the placebo group, only weak FGF-2 staining was observed, PDGF and IκB were not detected, and NF-κB was strongly observed in both cytoplasm and nuclei. CONCLUSIONS: Our findings suggest that DLE treatment modifies the OA process, promoting the expression of anti-inflammatory cytokines and diminishing the inflammatory effects, avoiding the nuclear translocation of NF-κB in chondrocytes.
Asunto(s)
Artritis Experimental/tratamiento farmacológico , Osteoartritis/tratamiento farmacológico , Factor de Transferencia/uso terapéutico , Animales , Artritis Experimental/metabolismo , Artritis Experimental/patología , Condrocitos/efectos de los fármacos , Condrocitos/metabolismo , Citocinas/metabolismo , Evaluación Preclínica de Medicamentos , Factor 2 de Crecimiento de Fibroblastos/metabolismo , Proteínas I-kappa B/metabolismo , Masculino , FN-kappa B/metabolismo , Osteoartritis/metabolismo , Osteoartritis/patología , Factor de Crecimiento Derivado de Plaquetas/metabolismo , Ratas Wistar , Factor de Transferencia/farmacologíaRESUMEN
1. Slow inotropic response following a sudden myocardium stretch seems to be an autocrine/paracrine mechanism the basis of which is not yet completely defined. 2. We compared the canine left ventricle (LV) response to sudden dilation when the LV was supported by the arterial blood of a support dog with when it was supported by an oxygenator + haemodialyser system. 3. A slow inotropic response (SIR) after dilation was seen in all six hearts supported by the donor dog, attaining 87 +/- 6% of immediate increase, whereas a mere 10% SIR occurred in only one out of seven hearts maintained by the oxygenator + haemodialyser. 4. These results indicate that SIR genesis involves one or more renewable components essential to the intracellular calcium gain elicited by stretch.
Asunto(s)
Dilatación Patológica/fisiopatología , Corazón/fisiopatología , Contracción Miocárdica/fisiología , Función Ventricular Izquierda/fisiología , Animales , Comunicación Autocrina/efectos de los fármacos , Comunicación Autocrina/fisiología , Presión Sanguínea/efectos de los fármacos , Soluciones para Diálisis/química , Soluciones para Diálisis/farmacología , Perros , Glucosa/administración & dosificación , Glucosa/farmacología , Corazón/efectos de los fármacos , Frecuencia Cardíaca/efectos de los fármacos , Técnicas In Vitro , Masculino , Contracción Miocárdica/efectos de los fármacos , Oxigenadores de Membrana , Comunicación Paracrina/efectos de los fármacos , Comunicación Paracrina/fisiología , Perfusión/métodos , Diálisis Renal/instrumentación , Diálisis Renal/métodos , Factor de Transferencia/química , Factor de Transferencia/farmacología , Trometamina/administración & dosificación , Trometamina/farmacología , Función Ventricular Izquierda/efectos de los fármacos , Presión Ventricular/efectos de los fármacos , Presión Ventricular/fisiologíaRESUMEN
OBJECTIVE: To investigate i) whether the Dialyzable Leukocyte Extract (DLE) modulates the production of proinflammatory cytokines in leukocytes activated by the bacterial cell wall components lipopolysaccharide (LPS), lipoteichoic acid (LTA), and peptidoglycan (PGN); ii) the effect of DLE on LPS-stimulated endothelial cells; and iii) whether the regulatory effect of DLE on inflammatory mediators is related to the modulation of Toll-like receptors (TLRs), NF-kappaB and cAMP signaling pathways. METHODS: Leukocytes were stimulated with LPS, LTA, and PGN in the presence of DLE. Endothelial cells were stimulated with LPS and treated with DLE. The levels of Tumor Necrosis Factor-alpha(TNFalpha), Interleukin-6 (IL-6), and IL-8 in culture supernatants were evaluated by ELISA. The expression of Toll-like receptor 2 (TLR2) and 4 (TLR4), NF-kappaB activity and cAMP levels were evaluated by flow cytometry, EMSA, and EIA, respectively. RESULTS: The addition of DLE to leukocytes stimulated with cell wall constituents suppressed the production of TNFalpha. However, DLE induced IL-8 release in monocytes and enhanced IL-6 and IL-8 production by activated monocytes and endothelial cells. Also, DLE induced TLR2 and TLR4 expression, and increased cAMP levels, whereas NF-kappaB activity was inhibited. CONCLUSIONS: The present data indicate the differential regulation by DLE of the production of TNFalpha, IL-6, and IL-8 cytokines, associated with effects on TLR2 and TLR4 expression and NF-kappaB and cAMP activities. We suggest a putative mechanism for the biological effects of DLE in activated leukocytes and endothelial cells.
Asunto(s)
Células Endoteliales/metabolismo , Mediadores de Inflamación/metabolismo , Leucocitos/metabolismo , Lipopolisacáridos/farmacología , Peptidoglicano/farmacología , Ácidos Teicoicos/farmacología , Factor de Transferencia/farmacología , Células Cultivadas , AMP Cíclico/metabolismo , Células Endoteliales/efectos de los fármacos , Humanos , Interleucina-6/biosíntesis , Interleucina-8/biosíntesis , Leucocitos/efectos de los fármacos , Glicoproteínas de Membrana/metabolismo , Monocitos/efectos de los fármacos , Monocitos/metabolismo , FN-kappa B/efectos de los fármacos , FN-kappa B/metabolismo , Receptores de Superficie Celular/metabolismo , Receptor Toll-Like 2 , Receptor Toll-Like 4 , Receptores Toll-Like , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
According to UNAIDS, the global HIV/AIDS epidemic increased to 40 million the number of people living with the virus around the world. Dialyzable leukocyte extract obtained by our group is a low molecular weight dialyzable material from peripheral human leukocytes previously in vitro induced with Sendai virus (DLE-ind), and more recently, from non-induced leukocytes (DLE n/i). Previous results have shown the ability of DLE-ind to inhibit HIV in vitro replication in MT4 cell; to reduce TNFalpha secretion, and to delay in vivo progression to AIDS in early stage of HIV infection. In this work we present evidences that DLE n/i also inhibits HIV in vitro replication and reduces TNFalpha secretion in human whole blood like DLE obtained from induced leukocytes. Taking together these results show that both properties of DLE, HIV in vitro inhibition and TNF production modulation, are not dependent on in vitro Sendai virus induction of leukocytes.
Asunto(s)
Células Sanguíneas/virología , Fibroblastos/virología , VIH-1/efectos de los fármacos , VIH-1/fisiología , Linfocitos T/virología , Factor de Transferencia/farmacología , Factor de Necrosis Tumoral alfa/metabolismo , Células Sanguíneas/efectos de los fármacos , Línea Celular , Relación Dosis-Respuesta a Droga , Fibroblastos/efectos de los fármacos , Humanos , Lipopolisacáridos/farmacología , Masculino , Linfocitos T/efectos de los fármacos , Replicación Viral/efectos de los fármacosRESUMEN
The pathophysiology of endotoxic shock is characterized by the activation of multiple pro-inflammatory genes and their products which initiate the inflammatory process. Endotoxic shock is a serious condition with high mortality. Bovine dialyzable leukocyte extract (bDLE) is a dialyzate of a heterogeneous mixture of low molecular weight substances released from disintegrated leukocytes of the blood or lymphoid tissue obtained from homogenized bovine spleen. bDLE is clinically effective for a broad spectrum of diseases. To determine whether bDLE improves survival and modulates the expression of pro-inflammatory cytokine genes in LPS-induced, murine endotoxic shock, Balb/C mice were treated with bDLE (1 U) after pretreatment with LPS (17 mg/kg). The bDLE improved survival (90%), suppressed IL-10 and IL-6, and decreased IL-1beta, TNF-alpha, and IL-12p40 mRNA expression; and decreased the production of IL-10 (P<0.01), TNF-alpha (P<0.01), and IL-6 (P<0.01) in LPS-induced, murine endotoxic shock. Our results demonstrate that bDLE leads to improved survival in LPS-induced endotoxic shock in mice, modulating the pro-inflammatory cytokine gene expression, suggesting that bDLE is an effective therapeutic agent for inflammatory illnesses associated with an unbalanced expression of pro-inflammatory cytokine genes such as in endotoxic shock, rheumatic arthritis and other diseases.
Asunto(s)
Endotoxinas/efectos adversos , Lipopolisacáridos/efectos adversos , Lipopolisacáridos/antagonistas & inhibidores , Choque Séptico/mortalidad , Choque Séptico/prevención & control , Factor de Transferencia/uso terapéutico , Animales , Bovinos , Citocinas/clasificación , Citocinas/genética , Citocinas/inmunología , Relación Dosis-Respuesta a Droga , Endotoxinas/antagonistas & inhibidores , Inflamación/genética , Inflamación/inmunología , Mediadores de Inflamación/inmunología , Mediadores de Inflamación/metabolismo , Inyecciones Intraperitoneales , Leucocitos/inmunología , México , Ratones , Ratones Endogámicos BALB C , Choque Séptico/inducido químicamente , Factor de Transferencia/farmacologíaRESUMEN
The human immunodeficiency virus type 1 (HIV-1) contains regulatory regions in its long terminal repeat (LTR) implicated in the control of viral gene expression. We previously demonstrated that Dialyzable Leukocyte Extract (DLE), a preparation derived from immune leukocytes, is able to inhibit HIV-1 replication in MT-4 cell cultures. Here, we examined the effect of DLE on the activation of NF-kappaB and Sp1 transcription factors. NF-kappaB activity was completely suppressed after seven days of treatment with 2.5 U/mL of DLE, with a parallel large reduction in the amounts of Sp1 complexes. These findings correlate with the maximum inhibitory effect on HIV-1 replication described in a previous report. IkappaBalpha and NF-kappaB p65(RelA) gene expression are not regulated by DLE in MT-4 cells. Although up to day, the precise molecular mechanism of DLE biological activity in HIV-1 infection remains unclear, this report presents data that indicate a potential downregulatory effect of DLE on HIV-1 gene expression.
Asunto(s)
Regulación Viral de la Expresión Génica/fisiología , VIH-1/genética , Factores de Transcripción/fisiología , Factor de Transferencia/farmacología , Secuencia de Bases , Línea Celular , Cartilla de ADN , Humanos , FN-kappa B/metabolismoRESUMEN
Dialysable leucocyte extract (DLE), obtained from lysed leucocytes, provide clinical effectiveness in a broad spectrum of diseases. Tumour necrosis factor (TNF) is raised in AIDS patients leading to an increase in human immunodeficiency virus (HIV) replication in vitro [1,2], whereas progression to AIDS in asymptomatic HIV infected individuals is retarded under treatment with DLE. In the present study we tested the DLE effect in vitro on both TNF biological activity (cytotoxicity) in L929 cells and its induction by lipopolysaccharide (LPS) in human monocytes as well as in whole blood from healthy donors. When monocytic cells were simultaneously exposed to LPS and DLE during a period of 5 1/2 hours, the induction of TNF was strongly diminished. The same inhibitory effect of DLE on TNF induction was observed when LPS was added to the culture medium prior to DLE. No significant effect of DLE on TNF-mediated cytotoxicity, even in the presence of the highest concentrations of DLE tested, was detected. DLE treatment of whole human blood regulates responses to LPS: simultaneous in vitro exposure to endotoxin provokes a remarkable decrease (4- and 1.6-fold) of TNF release. In pre-incubation experiments, TNF production was largely reduced or completed abrogated. These results could, in part, explain the in vivo observed effect, when under treatment with this extract, the progression to AIDS of HIV-infected individuals was retarded. The results suggest that "natural' substances like DLE may be important immunomodulators in inflammatory diseases.
Asunto(s)
Leucocitos/efectos de los fármacos , Leucocitos/metabolismo , Lipopolisacáridos/farmacología , Factor de Necrosis Tumoral alfa/metabolismo , Animales , Extractos Celulares/farmacología , Células Cultivadas , Citotoxicidad Inmunológica , Diálisis , Fibroblastos/efectos de los fármacos , Fibroblastos/inmunología , Humanos , Leucocitos/química , Activación de Linfocitos , Ratones , Factor de Transferencia/farmacología , Factor de Necrosis Tumoral alfa/biosíntesisRESUMEN
A group of 90 external asthmatics with cellular immunodeficiency or not was studied and treated during 10 months with transfer factor or double blind placebo. Total immunoglobulin serum studies (A, G, M and E), spontaneous rosette and intradermal tests, were made a month before the treatment's beginning and a month after the ending of the treatment. The patients were clinically evaluated every day in accordance to the intensity and the frequency of their crisis and with the immunological point of view of the tests made. There was not significant differences between the study groups treated with the transfer factor or not. Adverse reactions were not noticed.
Asunto(s)
Adyuvantes Inmunológicos/uso terapéutico , Asma/terapia , Factor de Transferencia/uso terapéutico , Adyuvantes Inmunológicos/efectos adversos , Adyuvantes Inmunológicos/farmacología , Adolescente , Adulto , Método Doble Ciego , Femenino , Volumen Espiratorio Forzado/efectos de los fármacos , Humanos , Inmunidad Celular/efectos de los fármacos , Huésped Inmunocomprometido , Inmunoglobulina E/sangre , Inmunoglobulinas/análisis , Pruebas Intradérmicas , Recuento de Linfocitos , Masculino , Persona de Mediana Edad , Formación de Roseta , Subgrupos de Linfocitos T/efectos de los fármacos , Factor de Transferencia/efectos adversos , Factor de Transferencia/farmacología , Resultado del TratamientoRESUMEN
The therapeutic panorama of immunomodulation and its effects on the modification of the immune reaction is reviewed. Particular reference is made to the transfer factor as a therapeutic element in bronchial asthma, which insures its efficacy or innocuity.