RESUMEN
Previous research has shown that the immunomodulatory peptide alpha-melanocyte-stimulating hormone (alpha-MSH) and its carboxy-terminal tripeptide KPV (Lys-Pro-Val alpha-MSH11-13) have antimicrobial influences. By inserting a Cys-Cys linker between two units of KPV, we designed the dimer [Ac-CKPV]2 that showed excellent candidacidal effects in pilot tests and was the subject of further investigations. [Ac-CKPV]2 was active against azole-resistant Candida spp. Therefore, the molecule appeared a promising candidate for therapy of fungal infections and was the subject of a structural study. 1H-NMR and restrained mechanic and dynamic calculations suggest that the peptide adopts an extended backbone structure with a beta-turn-like structure. These results open a pathway to development of additional novel compounds that have candidacidal effects potentially useful against clinical infections.
Asunto(s)
Antifúngicos/farmacología , Candida/efectos de los fármacos , alfa-MSH/química , Espectroscopía de Resonancia Magnética , Oligopéptidos/química , Conformación Proteica , alfa-MSH/análogos & derivados , alfa-MSH/farmacologíaAsunto(s)
Cromosomas Humanos Par 21 , Amplificación de Genes , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Proteínas Proto-Oncogénicas , Adolescente , Factores de Edad , Niño , Preescolar , Subunidad alfa 2 del Factor de Unión al Sitio Principal , Análisis Citogenético , Proteínas de Unión al ADN/genética , Femenino , Humanos , Masculino , Estudios Prospectivos , Factores de Transcripción/genéticaRESUMEN
Diamond-Blackfan anemia (DBA) is a heterogeneous genetic disorder characterized by red cell aplasia and congenital anomalies. One of what appears to be multiple DBA genes has been cloned. Affected individuals in the same family may vary dramatically as to the degree of anemia, response to corticosteroids, and the presence of congenital anomalies. The epidemiology of DBA has been gleaned largely from literature reviews. This approach is limited because of the two-fold disadvantage of the reporting bias inherent in the literature and the lack of an active patient database. The Diamond Blackfan Anemia Registry of North America (DBAR) is designed to overcome these disadvantages to study the epidemiology and biology of DBA. The DBAR is a comprehensive database of patients with DBA who are enrolled after informed consent is obtained. Identification of patients is made through outreach to pediatric and adult hematologists and the Diamond Blackfan Anemia Foundation. The patients and/or their families complete a detailed questionnaire. A review of medical records and telephone interviews are performed to complete and clarify the information provided. To date, 354 patients have been enrolled in the DBAR. Using this database, important epidemiologic, clinical, and laboratory observations have been made with regard to the clinical presentation, the inheritance of DBA, the genetics of congenital malformations, the therapeutic outcome, including the efficacy of hematopoietic stem cell transplantation, and the recognition of DBA as a cancer predisposition syndrome. In particular, the database is an essential substrate for DBA gene discovery.
Asunto(s)
Anemia de Fanconi/epidemiología , Niño , Preescolar , Anemia de Fanconi/genética , Anemia de Fanconi/terapia , Femenino , Trasplante de Células Madre Hematopoyéticas , Humanos , Lactante , Recién Nacido , Masculino , Linaje , Sistema de RegistrosRESUMEN
Thiopental is one of the intravenous anesthetics used widely. Several reports have demonstrated that thiopental inhibits the immune responses. We investigated whether or not thiopental inhibits the production of tumor necrosis factor-alpha (TNF-alpha) induced by lipopolysaccharide (LPS) in human glioma cells (A-172). Moreover, we determined whether or not thiopental modulates activation of the transcription factor NF-kappaB, a factor that regulates expression of the genes that code for proinflammatory cytokines in A-172 cells and in experimental murine brain inflammation. Thiopental inhibited TNF-alpha production induced by LPS in A-172 cells. Electrophoretic mobility shift assays demonstrated that thiopental inhibited NF-kappaB activation induced by LPS in A-172 cells. In experimental murine brain inflammation induced by intracerebroventricular injection of LPS, intraperitoneal injection of thiopental inhibited NF-kappaB activation. Western blot analysis indicated that this inhibition was linked to preservation of IkappaBalpha protein expression in A-172 cells. The chloramphenicol acetyltransferase assay revealed that NF-kappaB-dependent reporter gene expression was suppressed in A-172 cells exposed to thiopental. These findings are consistent with the idea that thiopental exerts antiinflammatory effects in cultured cells and experimental murine brain inflammation, through suppression of TNF-alpha production via inhibition of NF-kappaB activation.
Asunto(s)
Anestésicos Intravenosos/farmacología , Encéfalo/efectos de los fármacos , Encefalitis/tratamiento farmacológico , Proteínas I-kappa B , FN-kappa B/efectos de los fármacos , Tiopental/farmacología , Células Tumorales Cultivadas/efectos de los fármacos , Factor de Necrosis Tumoral alfa/efectos de los fármacos , Animales , Encéfalo/inmunología , Encéfalo/fisiopatología , Neoplasias Encefálicas , Proteínas de Unión al ADN/metabolismo , Encefalitis/inducido químicamente , Encefalitis/metabolismo , Regulación de la Expresión Génica/efectos de los fármacos , Regulación de la Expresión Génica/fisiología , Genes Reporteros/efectos de los fármacos , Genes Reporteros/fisiología , Glioma , Humanos , Sistema Inmunológico/efectos de los fármacos , Sistema Inmunológico/metabolismo , Lipopolisacáridos/farmacología , Masculino , Ratones , Ratones Endogámicos BALB C , Inhibidor NF-kappaB alfa , FN-kappa B/genética , FN-kappa B/metabolismo , Células Tumorales Cultivadas/inmunología , Células Tumorales Cultivadas/metabolismo , Factor de Necrosis Tumoral alfa/metabolismoRESUMEN
Lymphomatoid papulosis (LyP) is a benign, self-healing, papular eruption that can wax and wane over the course of time. Transformation to T-cell lymphoma has been well documented in 10% to 20% of adults with LyP, but there are have been no cases reported in patients younger than age 26 years. We describe the first pediatric patient, a 16-year-old girl, who had clinical features of LyP and concurrently was found to have a lesion diagnosed as Ki-1+ anaplastic large cell lymphoma. After treatment with chemotherapy, she has been in continuous remission for 16 months.
Asunto(s)
Linfoma Anaplásico de Células Grandes/complicaciones , Papulosis Linfomatoide/complicaciones , Adolescente , Protocolos de Quimioterapia Combinada Antineoplásica/administración & dosificación , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Biomarcadores de Tumor , Células Clonales/química , Células Clonales/patología , Ciclofosfamida/administración & dosificación , Doxorrubicina/administración & dosificación , Femenino , Reordenamiento Génico de la Cadena gamma de los Receptores de Antígenos de los Linfocitos T , Humanos , Linfoma Anaplásico de Células Grandes/tratamiento farmacológico , Linfoma Anaplásico de Células Grandes/patología , Papulosis Linfomatoide/tratamiento farmacológico , Papulosis Linfomatoide/patología , Invasividad Neoplásica , Células Madre Neoplásicas/química , Células Madre Neoplásicas/patología , Prednisona/administración & dosificación , Inducción de Remisión , Hombro , Vincristina/administración & dosificaciónRESUMEN
We report on maternal first cousins with bilateral microtia, micrognathia, cleft palate and hematologic findings of Diamond-Blackfan anemia (DBA). The similarity of findings shared between our cases and a female reported by Hasan and Inoue [1993] suggests that this is a distinctive syndrome, rather than a chance association. DBA is a heterogeneous disorder, caused in about 25% of cases by heterozygous mutations in the RPS19 gene (DBA1). Mutation analysis in our cases did not show an RPS19 mutation, and 2 alleles were present in each. Segregation analysis for DBA1 on chromosome 19 and DBA2 on 8p23 was not consistent with linkage. We conclude that this syndrome of microtia, cleft palate and DBA is not allelic to known DBA loci.
Asunto(s)
Anomalías Múltiples/patología , Fisura del Paladar/patología , Oído Externo/anomalías , Anemia de Fanconi/patología , Anomalías Múltiples/genética , Niño , Preescolar , Salud de la Familia , Femenino , Humanos , Lactante , Masculino , Linaje , Proteínas Ribosómicas/genéticaRESUMEN
The Diamond Blackfan Anemia (DBA) Registry of North America is a detailed database of patients with DBA from the United States and Canada. To date, 354 patients have been registered. From this database an analysis of the outcome of hematopoietic stem cell transplantation for DBA was undertaken. Of the 20 transplanted patients who met criteria for the diagnosis of DBA, eight underwent an allogeneic HLA-matched sibling hematopoietic stem cell transplant (SCT) and 12 an alternative donor SCT. The median age at transplant for all patients was 6 years 2 months; 3 years 10 months vs 9 years 1 month for HLA-matched sibling and alternative donor SCT, respectively. All of the HLA-matched sibling transplants were done using a non-irradiation-containing regimen, whereas the majority of alternative donor transplants were performed using total body irradiation. The survival for HLA-matched sibling vsalternative donor transplant was 87.5% +/- 11.7% vs 14.1% +/- 12.1% at greater than 5 years from SCT (P = 0.015). The use of HLA-matched sibling SCT should be considered for all patients with suitable donors. However, alternative donor SCT in DBA must be approached cautiously, the potential for severe aplastic anemia (SAA) or hematopoietic malignancy not withstanding.
Asunto(s)
Anemia de Fanconi/terapia , Trasplante de Células Madre Hematopoyéticas/normas , Trasplante Homólogo/métodos , Adolescente , Adulto , Causas de Muerte , Niño , Preescolar , Anemia de Fanconi/complicaciones , Anemia de Fanconi/mortalidad , Enfermedad Injerto contra Huésped/etiología , Trasplante de Células Madre Hematopoyéticas/efectos adversos , Trasplante de Células Madre Hematopoyéticas/métodos , Prueba de Histocompatibilidad , Humanos , Lactante , Núcleo Familiar , Sistema de Registros , Tasa de Supervivencia , Acondicionamiento Pretrasplante/métodos , Trasplante Homólogo/efectos adversos , Trasplante Homólogo/normas , Resultado del TratamientoRESUMEN
Diamond-Blackfan anemia (DBA) is a rare congenital hypoplastic anemia that usually presents early in infancy and is inherited in 10% to 20% of cases. Linkage analysis has shown that DBA in many of both dominant and recessive DBA families mapped to chromosome 19q13.2 leading to the cloning of a gene on chromosome 19q13.2 that encodes a ribosomal protein, RPS19. However, subsequently, mutations of the RPS19 gene have only been identified in 25% of all patients with DBA. This study analyzed 14 multiplex DBA families, 9 of which had 19q13.2 haplotypes inconsistent with 19q linkage. A genome-wide search for linked loci suggested the presence of a second DBA locus in a 26.4-centimorgan (cM) interval on human chromosome 8p. Subsequently, 24 additional DBA families were ascertained and all 38 families were analyzed with additional polymorphic markers on chromosome 8p. In total, 18 of 38 families were consistent with linkage to chromosome 8p with a maximal LOD score with heterogeneity of 3.55 at D8S277 assuming 90% penetrance. The results indicate the existence of a second DBA gene in the 26.4-cM telomeric region of human chromosome 8p23.3-p22, most likely within an 8.1-cM interval flanked by D8S518 and D8S1825. Seven families were inconsistent with linkage to 8p or 19q and did not reveal mutations in the RPS19 gene, suggesting further genetic heterogeneity. (Blood. 2001;97:2145-2150)
Asunto(s)
Cromosomas Humanos Par 8/genética , Anemia de Fanconi/genética , Heterogeneidad Genética , Cromosomas Humanos Par 19/genética , Cromosomas Humanos Par 8/ultraestructura , Análisis Mutacional de ADN , Femenino , Marcadores Genéticos , Pruebas Genéticas , Haplotipos/genética , Humanos , Escala de Lod , Masculino , Linaje , FenotipoRESUMEN
PURPOSE: Diamond-Blackfan anemia (DBA) is a congenital pure red cell aplasia, usually presenting in infancy or early childhood. A review of the literature strongly supports a predisposition to hematopoietic malignancy. Recently, solid tumors have been reported, some attributable to hemosiderosis and/or androgen therapy. Two cases of osteogenic sarcoma have also been documented. An analysis from the Diamond-Blackfan Anemia Registry was performed to evaluate the cancer risk in patients with DBA. METHODS: The Diamond-Blackfan Anemia Registry of North America (DBAR) is a comprehensive database of patients with DBA enrolled, after informed consent, through outreach to pediatric hematologists and family groups. The patients and/or their families complete a detailed questionnaire, and a review of medical records and telephone interviews are performed to complete and clarify the information provided. RESULTS: Of the 354 patients registered in the DBAR, there were six patients meeting the accepted diagnostic criteria for DBA who were found to have malignancies. Three patients had osteogenic sarcoma diagnosed, one with myelodysplastic syndrome, one with colon carcinoma, and one with a soft tissue sarcoma. CONCLUSION: There appears to be an association of osteogenic sarcoma with DBA. A young age at presentation may be a feature of DBA-associated osteogenic sarcoma. Because of the immaturity of the database, the actuarial risk for osteogenic sarcoma and other cancers in individuals with DBA cannot be ascertained. Speculation is made regarding the nature of the molecular defect leading to the association of DBA and osteogenic sarcoma.
Asunto(s)
Neoplasias Óseas/epidemiología , Anemia de Fanconi/epidemiología , Osteosarcoma/epidemiología , Sistema de Registros , Adolescente , Adulto , Neoplasias Óseas/complicaciones , Preescolar , Neoplasias del Colon/complicaciones , Bases de Datos Factuales , Susceptibilidad a Enfermedades , Anemia de Fanconi/complicaciones , Femenino , Humanos , Masculino , Síndromes Mielodisplásicos/complicaciones , Osteosarcoma/complicaciones , Factores de Riesgo , Sarcoma/complicaciones , Estados UnidosRESUMEN
Thrombocytopenia absent corpus callosum, characterized by refractory thrombocytopenia, agenesis of the corpus callosum, hypoplastic cerebellum, abnormal facies, and developmental delay, represents a relatively newly described clinical entity. An 18-month-old girl with agenesis of the corpus callosum, hypoplasia of the cerebellar vermis, hypotonia, and severe developmental delay presented with thrombocytopenia. She had a distinctive facies with microcephaly, broad nasal root with upturned nose, small upper lip, and micrognathia. A bone marrow aspirate and biopsy showed normal cellularity with dysplastic megakaryocytes. Thrombocytopenia absent corpus callosum is compared with other conditions characterized by congenital non-immune thrombocytopenia.
Asunto(s)
Anomalías Múltiples/diagnóstico , Agenesia del Cuerpo Calloso , Discapacidades del Desarrollo/diagnóstico , Cara/anomalías , Trombocitopenia/congénito , Adulto , Niño , Preescolar , Femenino , Humanos , Lactante , Masculino , Síndrome , Tomografía Computarizada por Rayos XRESUMEN
alpha-Melanocyte-stimulating hormone (alpha-MSH) modulates inflammation. We investigated the influence of alpha-MSH on NF-kappaB activation in human pulmonary epithelial cells (A549) using a plasmid vector encoding alpha-MSH (pCMV-ssMSH). Electrophoretic mobility shift assays demonstrated that NF-kappaB activation induced by lipopolysaccharide was inhibited in A549 cells transfected with pCMV-ssMSH. Western blot analysis revealed that this inhibition was linked to preservation of expression of IkappaBalpha protein. Chloramphenicol acetyltransferase assay indicated that NF-kappaB-dependent reporter gene expression was suppressed in A549 cells transfected with pCMV-ssMSH. The findings indicate that anti-inflammatory actions are exerted via modulation of NF-kappaB activation by preservation of IkappaBalpha protein in human pulmonary epithelial cells transfected with alpha-MSH vector. We showed a possibility of gene therapy for chronic inflammatory lung diseases.
Asunto(s)
Células Epiteliales/metabolismo , Proteínas I-kappa B , Pulmón/metabolismo , FN-kappa B/antagonistas & inhibidores , alfa-MSH/metabolismo , Secuencia de Aminoácidos , Secuencia de Bases , Western Blotting , Línea Celular , Medios de Cultivo Condicionados , Proteínas de Unión al ADN/análisis , Proteínas de Unión al ADN/metabolismo , Células Epiteliales/efectos de los fármacos , Regulación de la Expresión Génica/efectos de los fármacos , Genes Reporteros , Terapia Genética , Vectores Genéticos , Humanos , Lipopolisacáridos/farmacología , Pulmón/citología , Pulmón/efectos de los fármacos , Datos de Secuencia Molecular , Inhibidor NF-kappaB alfa , FN-kappa B/metabolismo , Proteínas Nucleares/metabolismo , Neumonía/genética , Neumonía/terapia , Transporte de Proteínas/efectos de los fármacos , Transfección , alfa-MSH/genéticaRESUMEN
The purpose of the present research was to determine if alpha-melanocyte-stimulating hormone (alpha-MSH) and its C-terminal tripeptide [alpha-MSH (11-13), KPV] alter HIV expression in infected cells. The results indicate that chronically HIV-1-infected promonocytic U1 cells produce alpha-MSH and that immunoneutralization of the endogenous peptide enhances HIV expression. Because U1 cells express the alpha-MSH receptor 1 (MC1R), an autocrine-inhibitory circuit based on the peptide and its receptor likely occurs in these cells. To determine effects of pharmacological concentrations of alpha-MSH peptides on HIV expression, we measured p24 antigen release by TNF-alpha-stimulated U1 cells exposed to a wide range of concentrations of synthetic alpha-MSH and KPV. Viral expression was reduced by both peptides. KPV also effectively reduced HIV replication in acutely infected monocyte-derived macrophages (MDM). The basis of the peptide influence on viral replication is at the transcriptional level; KPV inhibited activation of NF-kappaB that is known to enhance viral expression. Endogenous alpha-MSH likely contributes to natural defense against HIV. However, greater concentrations of synthetic peptide are much more effective in reducing HIV expression in infected cells.
Asunto(s)
VIH-1/efectos de los fármacos , Hormonas Estimuladoras de los Melanocitos/farmacología , Monocitos/virología , Fragmentos de Péptidos/farmacología , Replicación Viral/efectos de los fármacos , alfa-MSH/análogos & derivados , alfa-MSH/farmacología , ADN Viral/metabolismo , Proteína p24 del Núcleo del VIH/metabolismo , VIH-1/metabolismo , VIH-1/fisiología , Humanos , Monocitos/efectos de los fármacos , Monocitos/metabolismo , FN-kappa B/metabolismo , Transcripción Genética/efectos de los fármacos , Factor de Necrosis Tumoral alfa/farmacología , alfa-MSH/biosíntesis , alfa-MSH/fisiologíaRESUMEN
PURPOSE: Ketamine reduces endotoxin-induced production of proinflammatory cytokines, including tumour necrosis factor-alpha (TNF), in several types of inflammatory cells, including monocytes and macrophages. Transcription of the genes that encode production of these proinflammatory cytokines is regulated by nuclear factor-kappa B (NF-kappaB). Cytoplasmic B protein is activated by endotoxin (LPS) as well as by TNF, allowing B protein to migrate into the cell nucleus to activate gene transcription for these inflammatory mediators. Because NF-kappaB is likely involved in brain injury and inflammatory neurodegenerative disease, such as multiple sclerosis, we examined whether ketamine inhibits LPS-induced activation of NF-kappaB in human glioma cells in vitro and intact mouse brain cells in vivo. METHODS: Endotoxin-induced NF-kappaB expression in both the human glioma cells in vitro and the intact mouse brain cells in vivo was determined by electrophoretic mobility shift assays (EMSA) of nuclear extracts and measurement of NF-kappaB expression by densitometry. Endotoxin was injected intracerebroventricularly in vivo and intact brain was harvested. Klenow fragment labeling was used to identify NF-kappaB protein for both the in vivo and vitro experiments. RESULTS: Endotoxin treatment increased NF-kappaB expression (P < 0.05) both in vivo and vitro compared with control (untreated) cells. Ketamine suppressed endotoxin-induced neuronal NF-kappaB activation in a dose-dependent manner (P < 0.05, except for the 10(-5) M concentration in vitro) both in vivo and vitro. CONCLUSION: Ketamine inhibits endotoxin-induced NF-kappaB expression in brain cells in vivo and vitro and it is suggested that this may have implications in the neuroprotective effects of ketamine reported by other investigators.
Asunto(s)
Encéfalo/efectos de los fármacos , Ketamina/farmacología , Lipopolisacáridos/toxicidad , FN-kappa B/metabolismo , Animales , Encéfalo/metabolismo , Relación Dosis-Respuesta a Droga , Humanos , Masculino , Ratones , Ratones Endogámicos BALB C , Óxido Nítrico/biosíntesis , Células Tumorales CultivadasRESUMEN
Over the past two decades, research in animal models has indicated that alpha-melanocyte-stimulating hormone (alpha-MSH) has potent anti-inflammatory properties. Furthermore, recent data show that the peptide has antimicrobial effects and probably contributes to innate immunity. alpha-MSH, which is produced by many extrapituitary human cells, should no longer be considered solely a pituitary hormone; rather, it should be viewed as a ubiquitous modulatory peptide.
Asunto(s)
alfa-MSH/fisiología , Animales , Humanos , Infecciones/metabolismo , Inflamación/metabolismo , alfa-MSH/metabolismoRESUMEN
BACKGROUND: Clinical and/or laboratory signs of systemic inflammation occur frequently in patients undergoing long-term haemodialysis. It is likely, therefore, that a compensatory release of endogenous anti-inflammatory molecules occurs to limit host reactions. The aim of the present research was to determine if the potent anti-inflammatory peptide alpha-melanocyte-stimulating hormone (alpha-MSH), a pro-opiomelanocortin derivative, is increased in plasma of haemodialysis patients. Because endotoxin and cytokines induce alpha-MSH in vivo and in vitro, we also measured plasma concentrations of endotoxin, interleukin-6 (IL-6), and tumour necrosis factor alpha (TNF-alpha), and the two circulating products of activated monocytes, nitric oxide (NO) and neopterin. METHODS: Thirty-five chronic haemodialysis patients, 20 patients with chronic renal failure not yet on dialysis, and 35 normal controls were included in the study. In the haemodialysis group, blood samples were obtained before and at the end of a dialysis session. Plasma alpha-MSH was measured using a double antibody radioimmunoassay, and IL-6, TNF-alpha, and neopterin using specific enzyme-linked immunosorbent assays. Plasma nitrites were determined by a colorimetric method, and endotoxin with the quantitative chromogenic LAL (limulus amoebocyte lysate) method. RESULTS: Mean plasma alpha-MSH was higher in haemodialysis patients than in control subjects, with the peptide concentrations being particularly elevated in dialysed patients with detectable endotoxin. High alpha-MSH concentrations were observed in the pre-dialysis samples, with no substantial change at the end of the dialysis session. Plasma concentrations of IL-6, TNF-alpha, neopterin, and NO were generally elevated in chronic haemodialysis patients and there was a negative correlation between circulating alpha-MSH and IL-6. In patients with renal failure not yet on dialysis, mean plasma alpha-MSH was similar to that of normal subjects. CONCLUSIONS: alpha-MSH is increased in the circulation of chronic haemodialysis patients and particularly so in case of detectable endotoxaemia. Reduction of renal clearance is unlikely to contribute to the observed rise of the peptide because alpha-MSH concentration is not increased in patients with chronic renal failure who are not yet on dialysis. It is likely that dialysis-associated endotoxaemia, directly and/or through cytokine release, enhances the production of the anti-inflammatory mediator alpha-MSH that limits host reactions.
Asunto(s)
Diálisis Renal , alfa-MSH/sangre , Adulto , Anciano , Endotoxinas/sangre , Femenino , Humanos , Mediadores de Inflamación/sangre , Interleucina-6/sangre , Fallo Renal Crónico/sangre , Fallo Renal Crónico/terapia , Masculino , Persona de Mediana Edad , Neopterin/sangre , Óxido Nítrico/sangre , Concentración Osmolar , Factores de Tiempo , Factor de Necrosis Tumoral alfa/análisisRESUMEN
OBJECTIVES: The aim of this research was to investigate endogenous concentrations and anti-cytokine effects of the antiinflammatory peptide alpha-melanocyte stimulating hormone (alpha-MSH) in patients with systemic inflammation. The objectives were to determine the following: changes over time of plasma alpha-MSH and relationship with patient outcome, correlation between plasma alpha-MSH and tumor necrosis factor (TNF)-alpha plasma concentration and production in whole blood samples, and influences of alpha-MSH on production of TNF-alpha and interleukin (IL)-1beta in whole blood samples stimulated with lipopolysaccharide (LPS). DESIGN: Prospective, nonrandomized, clinical study. SETTING: Intensive care unit of a university hospital. PATIENTS: A total of 21 patients with sepsis syndrome/septic shock and an equal number of healthy volunteers. INTERVENTIONS: Circulating alpha-MSH and TNF-alpha concentrations and TNF-alpha production in supernatants of LPS (1 ng/mL)-stimulated whole blood were measured repeatedly. To determine whether alpha-MSH can modulate production of TNF-alpha and IL-1 beta, these cytokines were measured in whole blood samples stimulated with LPS (1 ng/mL) in the presence or absence of concentrations of the peptide. MEASUREMENTS AND MAIN RESULTS: Plasma alpha-MSH was low in early samples and gradually increased in patients who recovered but not in those who died. There was a negative correlation between plasma concentrations of alpha-MSH and TNF-alpha. In blood samples taken at early phases of sepsis syndrome, production of TNF-alpha was reduced relative to control values; such production increased in patients who recovered but not in those who died. Addition of alpha-MSH to LPS-stimulated whole blood samples inhibited production of TNF-alpha and IL-1beta in a concentration-dependent manner. CONCLUSIONS: In patients with systemic inflammation, there are substantial changes over time in plasma concentrations of alpha-MSH that are reduced in early phases of the disease. Reduction of this endogenous modulator of inflammation could be detrimental to the host. Addition of alpha-MSH to LPS-stimulated blood samples reduces production of cytokines involved in development of septic syndrome. This inhibition by alpha-MSH, a peptide that is beneficial in treatment of experimental models of sepsis, might therefore be useful to treat sepsis syndrome in humans.
Asunto(s)
Citocinas/antagonistas & inhibidores , Choque Séptico/inmunología , Síndrome de Respuesta Inflamatoria Sistémica/inmunología , alfa-MSH/sangre , Adulto , Anciano , Citocinas/inmunología , Femenino , Mortalidad Hospitalaria , Humanos , Interleucina-1/antagonistas & inhibidores , Interleucina-1/inmunología , Lipopolisacáridos/inmunología , Masculino , Persona de Mediana Edad , Pronóstico , Choque Séptico/mortalidad , Tasa de Supervivencia , Síndrome de Respuesta Inflamatoria Sistémica/mortalidad , Factor de Necrosis Tumoral alfa/antagonistas & inhibidores , Factor de Necrosis Tumoral alfa/inmunologíaRESUMEN
Sodium valproate (VPA) is frequently used to treat epilepsy and convulsive disorders. Several reports have indicated that anti-epileptic drugs (AED) affect the immune system, but the mechanism has not been clear. We examined whether the commonly used AEDs, diazepam (DZP), carbamazepine (CBZ), phenobarbital (PB), phenytoin (PHT), and VPA, can inhibit activation of the nuclear transcription factor kappa B (NF-kappaB), in human monocytic leukemia cells (THP-1) and in human glioma cells (A-172). NF-kappaB is essential to the expression of the kappa light chain of immunoglobulin and proinflammatory cytokines. Electrophoretic mobility shift assays (EMSA) of nuclear extracts demonstrated that VPA inhibits NF-kappaB activation induced by lipopolysaccharide (LPS), but the other AEDs do not. Western blot analysis revealed that this inhibition is not linked to preservation of expression of IkappaBalpha protein. Chloramphenicol acetyltransferase (CAT) assay indicated that NF-kappaB-dependent reporter gene expression is suppressed in glioma cells pretreated with VPA. VPA significantly inhibited LPS-induced production of TNF-alpha and IL-6 by THP-1 cells, whereas other AEDs did not. The findings are consistent with the idea that VPA suppresses TNF-alpha and IL-6 production via inhibition of NF-kappaB activation. Our results suggest that VPA can modulate immune responses in vitro. These findings raise the possibility that such modulation might occur with clinical use of VPA.
Asunto(s)
Anticonvulsivantes/farmacología , Interleucina-6/biosíntesis , FN-kappa B/efectos de los fármacos , FN-kappa B/metabolismo , Factor de Necrosis Tumoral alfa/biosíntesis , Ácido Valproico/farmacología , Células Cultivadas , Humanos , Lipopolisacáridos/farmacología , Factor de Necrosis Tumoral alfa/efectos de los fármacosRESUMEN
The presence of the ancient anti-inflammatory peptide alpha-melanocyte-stimulating hormone [alpha-MSH (1-13), SYSMEHFRWGKPV] in barrier organs such as gut and skin suggests a role in the nonspecific (innate) host defense. alpha-MSH and and its carboxy-terminal tripeptide (11-13, KPV) were determined to have antimicrobial influences against two major and representative pathogens: Staphylococcus aureus and Candida albicans. alpha-MSH peptides significantly inhibited S. aureus colony formation and reversed the enhancing effect of urokinase on colony formation. Antimicrobial effects occurred over a broad range of concentrations including the physiological (picomolar) range. Small concentrations of alpha-MSH peptides likewise reduced viability and germ tube formation of the yeast C. albicans. Antimicrobial influences of alpha-MSH peptides could be mediated by their capacity to increase cellular cAMP. Indeed, this messenger was significantly augmented in peptide-treated yeast and the potent adenylyl cyclase inhibitor dideoxyadenosine (ddAdo) partly reversed the killing activity of alpha-MSH peptides. Reduced killing of pathogens is a detrimental consequence of therapy with anti-inflammatory drugs. Because alpha-MSH has potent anti-inflammatory effects we determined influences of alpha-MSH on C. albicans and S. aureus killing by human neutrophils. alpha-MSH peptides did not reduce killing but rather enhanced it, likely as a consequence of the direct antimicrobial activity. alpha-MSH peptides that combine antipyretic, anti-inflammatory, and antimicrobial effects could be useful in treatment of disorders in which infection and inflammation coexist.
Asunto(s)
Antibacterianos/farmacología , Antifúngicos/farmacología , Candida albicans/efectos de los fármacos , Fragmentos de Péptidos/farmacología , Staphylococcus aureus/efectos de los fármacos , alfa-MSH/farmacología , Antibacterianos/administración & dosificación , Antibacterianos/química , Antifúngicos/administración & dosificación , Antifúngicos/química , AMP Cíclico/fisiología , Citotoxicidad Inmunológica , Relación Dosis-Respuesta a Droga , Humanos , Neutrófilos/fisiología , Fragmentos de Péptidos/administración & dosificación , Fragmentos de Péptidos/química , Sistemas de Mensajero Secundario/efectos de los fármacos , Relación Estructura-Actividad , Activador de Plasminógeno de Tipo Uroquinasa/antagonistas & inhibidores , alfa-MSH/administración & dosificación , alfa-MSH/análogos & derivados , alfa-MSH/químicaRESUMEN
Inflammation in peripheral tissues should benefit from a novel focus on descending neuroimmunomodulatory anti-inflammatory influences stemming from CNS receptors for the peptide alpha-MSH. Because activation of glial and peripheral cell receptors modulates inflammatory activity, alpha-MSH peptides may aid treatment of a variety of CNS and peripheral inflammatory disorders. Activation of melanocortin receptors should thus promote control of inflammatory diseases.