Anti-melanocortin-4 receptor autoantibodies in obesity.

BACKGROUND: The melanocortin-4 receptor (MC4R) is part of an important pathway regulating energy balance. Here we report the existence of autoantibodies (autoAbs) against the MC4R in sera of obese patients. METHODS: The autoAbs were detected after screening of 216 patients' sera by using direct and inhibition ELISA with an N-terminal sequence of the MC4R. Binding to the native MC4R was evaluated by flow cytometry, and pharmacological effects were evaluated by measuring adenylyl cyclase activity. RESULTS: Positive results in all tests were obtained in patients with overweight or obesity (prevalence, 3.6%) but not in normal weight patients. The selective binding properties of anti-MC4R autoAbs were confirmed by surface plasmon resonance and by immunoprecipitation with the native MC4R. Finally, it was demonstrated that these autoAbs increased food intake in rats after passive transfer via intracerebroventricular injection. CONCLUSION: These observations suggest that inhibitory anti-MC4R autoAbs might contribute to the development of obesity in a small subpopulation of patients.

Expression of CD34 in pulmonary endothelial cells in vivo.

OBJECTIVES: The morphological phenotype of endothelial cells (EC) is specific for individual types of vessels. There are, however, no studies on the phenotypical diversity of EC in human lung tissue. The influences exerted by physiological factors (e.g. age, sex) or pathological factors (e.g. pulmonary hypertension) on EC have not been ascertained up to now. METHODS: In order to determine the influence of pulmonary hypertension (PH), age and sex on EC, we localized the expression of the endothelial marker CD34 immunohistologically by light microscopy and laser scanning microscopy in lung tissue specimens from children and adults. RESULTS: Capillary EC showed a stronger staining reaction than EC in arteries, veins, arterioles or venules. The staining intensity increased with age in veins and arteries and decreased with age in venules, arterioles and capillaries. Sex exerted no statistically significant influence. CD34 was generally more strongly expressed in specimens with PH than in those without. CONCLUSION: The study demonstrates for the first time that (1) CD34 is heterogeneously expressed by human pulmonary EC, and (2) the physiological/pathophysiological factors age/PH influence CD34 expression. Hence a correlation between CD34 expression and its role as adhesion molecule and a link between CD34 expression and maturation are subject of discussion.

Expression of the endothelial markers PECAM-1, vWf, and CD34 in vivo and in vitro.

EC culture models are essential to study pathological alterations of endothelial cells (ECs) in pulmonary vascular diseases under standardized conditions. Nevertheless, little is known about the spectrum of alterations of vessel-specific endothelial phenotypes in monolayer cultures. For the comparative study of endothelial markers in vivo and in vitro we investigated immunohistochemically the expression of PECAM-1, vWf, and CD34 by pulmonary ECs in vivo and in stimulated/unstimulated human umbilical vein endothelial cells (HU-VEC) and human pulmonary microvascular endothelial cells (HPMEC). In vivo, vessel type-specific expression patterns were found for vWf and CD34, while PECAM-1 was homogeneously and strongly expressed. While all HUVEC showed a marked vWf staining, about two-thirds of HPMEC exhibited a strong and the rest a moderate vWf staining. In both in vitro models all ECs were clearly PECAM-1-positive. However, only about 20% of the HUVEC and HPMEC were CD34-positive. Our results demonstrate the reduced expression of vessel type-specific endothelial phenotypes by endothelial monolayer cultures, stressing the need to improve culture conditions as well as develop cocultures and three-dimensional culture models. Moreover, the need for endothelial markers specific for single microvascular type ECs becomes obvious in order to establish cultures consisting of only one microvascular ECs subpopulation.

Correlation of age with in vivo expression of endothelial markers.

The importance of endothelial senescence as a pathogenetic factor in age-related vascular alterations has almost exclusively been studied in vitro. However, the in vitro-findings have rarely been compared with histomorphological changes in aging human tissue or in age-related degenerative diseases. Therefore, we compared the expression of the endothelial marker CD34 and the procoagulant protein von Willebrand factor (vWf) in lung endothelium by conventional immunohistochemistry and confocal laser scan microscopy (CLSM), taking the age of the patients into consideration. The staining reactions were statistically analysed by covariance analysis. With age the endothelial staining intensity of CD34 increased in arteries and veins, but decreased in arterioles, capillaries and venules. For vWf, on the contrary, the endothelial staining intensity increased with age in all types of vessels. CLSM confirmed a mosaic staining pattern. This study demonstrates age-associated phenotypical alterations of CD34 and vWf. Whether the down-regulation of CD34 correlates with an age-associated reduction of the angiogenetic properties of EC or an age-related over-expression of vWf as a relevant cofactor for the raised coagulatory activity and the increase in thrombotic diseases resp coronary heart disease in older patients, remains subject to debate.