RESUMEN
BACKGROUND AND AIMS: Patients suffering from inflammatory bowel disease show increased levels of the mast cell products histamine and tumour necrosis factor alpha (TNF-alpha). Treating these patients with antibodies against TNF-alpha diminishes the symptoms of diarrhoea. In this study, the effect of TNF-alpha on ion secretion induced by the mast cell mediator histamine in HT29cl.19A cells and mouse distal colon was investigated and the possible second messengers involved were studied. METHODS: Electrophysiology of filter grown HT29cl.19A cells and isolated mouse distal colon was used to monitor the secretory response to histamine with and without prior exposure to TNF-alpha for 3-24 hours. Phospholipase D (PLD) activity and phosphatidic acid levels were analysed by 32P(i) labelling of HT29cl.19A cells. RESULTS: In both experimental systems TNF-alpha was found to potentiate ion secretion induced by histamine. Phospholipid analysis of HT29cl.19A cells revealed that histamine activates the PLD pathway. Furthermore, TNF-alpha pretreated cells were found to have decreased phosphatidic acid levels, the intermediate product of the PLD pathway, which indicates upregulation of the enzyme phosphatidic acid phosphatase. CONCLUSIONS: The mast cell products TNF-alpha and histamine synergistically stimulate ion secretion in intestinal epithelium via upregulation of the PLD pathway.
Asunto(s)
Canales de Cloruro/efectos de los fármacos , Colon/efectos de los fármacos , Histamina/farmacología , Fosfolipasa D/fisiología , Factor de Necrosis Tumoral alfa/farmacología , Animales , Línea Celular , Células Cultivadas , Canales de Cloruro/metabolismo , Colon/fisiología , Sinergismo Farmacológico , Electrofisiología , Femenino , Humanos , Mucosa Intestinal/efectos de los fármacos , Mucosa Intestinal/fisiología , Ratones , Ratones Endogámicos BALB C , Ácidos Fosfatidicos/análisisRESUMEN
In a previous study, it was found that exposure to tumor necrosis factor-alpha (TNF-alpha) potentiated the electrophysiological response to carbachol in a time-dependent and cycloheximide-sensitive manner. It was deduced that the potentiation could be due to protein kinase C activity because of increased 1,2-diacylglycerol. It was also observed that propranolol could decrease the electrophysiological response to carbachol (Oprins JC, Meijer HP, and Groot JA. Am J Physiol Cell Physiol 278: C463-C472, 2000). The aim of the present study was to investigate whether the phospholipase D (PLD) pathway plays a role in the carbachol response and the potentiating effect of TNF-alpha. The transphosphatidylation reaction in the presence of the primary alcohol 1-butanol [leading to stable phosphatidylbutanol (Pbut) formation] was used to measure activity of PLD. The phosphatidic acid (PA) levels were also measured. Muscarinic stimulation resulted in an increased formation of Pbut and PA. TNF-alpha decreased levels of PA.
Asunto(s)
Carbacol/farmacología , Cloruros/metabolismo , Agonistas Colinérgicos/farmacología , Fosfolipasa D/metabolismo , Factor de Necrosis Tumoral alfa/metabolismo , 1-Butanol/farmacología , Antagonistas Adrenérgicos beta/farmacología , Carcinógenos/farmacología , Cromatografía en Capa Delgada , Diacilglicerol Quinasa/antagonistas & inhibidores , Diacilglicerol Quinasa/metabolismo , Diglicéridos/metabolismo , Relación Dosis-Respuesta a Droga , Inhibidores Enzimáticos/farmacología , Células HT29 , Humanos , Indoles/farmacología , Mucosa Intestinal/enzimología , Maleimidas/farmacología , Ésteres del Forbol/farmacología , Propranolol/farmacología , Proteína Quinasa C/antagonistas & inhibidores , Proteína Quinasa C/metabolismo , Pirimidinonas/farmacología , Receptores Muscarínicos/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/fisiología , Tiazoles/farmacología , Factor de Necrosis Tumoral alfa/farmacologíaRESUMEN
A comparison was made between the diagnostic value of assaying nickel-induced lymphocyte proliferation (lymphocyte transformation test, LTT) and migration inhibition factor (MIF) production in nickel contact sensitivity. Although lymphocyte proliferation was significantly increased in the group of patients with skin test reactivity to nickel, positive LTT were also frequently found in skin test-negative subjects: in 63% of subjects with and in 30% of subjects without a history of metal allergy. This would limit the value of the LTT as an in vitro correlate of skin test reactivity. However, in certain patients positive lymphocyte transformation may reveal nickel sensitization at a time of undetectable skin reactivity. Data obtained with the macrophage migration inhibition test (MMIT) showed a good correlation with nickel patch test reactions. Accurate determination of MIF became feasible by using cells from the human monocytoid cell line U937 as target cells in a microdroplet agarose assay. Using this MMIT, positive reactions occurred in 13% of the healthy controls and false-negative reactions were found in 26% of patients with positive skin test reactivity to nickel. As LTT and MMIT data appeared to be only weakly correlated in the individuals tested, a dual parameter analysis was performed. An excellent correlation [p = 1.8 (10(-8]] was found between skin test and in vitro reactivity for individuals with matching in vitro results (60% of all individuals tested). In those individuals with discordant in vitro data, skin testing will remain indispensable for diagnosing nickel allergy.
Asunto(s)
Dermatitis por Contacto/diagnóstico , Níquel/efectos adversos , Línea Celular , Dermatitis por Contacto/etiología , Humanos , Leucemia Experimental/inmunología , Leucemia Experimental/patología , Activación de Linfocitos , Factores Inhibidores de la Migración de Macrófagos/biosíntesis , Pruebas CutáneasRESUMEN
A prospective study of the development of hand eczema was initiated in 86 junior hairdressers and 217 junior nurses. Data obtained at the start of their apprenticeships are presented. None of the junior apprentices presented with hand eczema, but a history of hand eczema was reported by 22/303 (7%) of the students. Almost half of this group (10/22, 45%) could be classified as atopics. Students without a history of hand eczema showed a similar frequency of atopy (17%) as observed in the general population. History of hand eczema was not related to nickel hypersensitivity, as assessed by patch testing. The incidence of nickel hypersensitivity was high in junior hairdressers (26%), compared to junior nurses (12%). Information as to previous contacts with nickel suggested that development of nickel allergy had been promoted by ear-piercing. A lower incidence of nickel hypersensitivity was observed if this potentially sensitizing event had been proceeded by orthodontic treatment with nickel-containing materials. This phenomenon is discussed in terms of orally-induced tolerance.