Bahn, R. S.

Heufelder, A. E.

Scriba, Peter Christian

English

Open Access LMU

European Journal of Endocrinology Formerly Acta Endocrinologica Advance Abstracts of Papers SCANDINAVIAN CONGRESS IN ENDOCRINOLOGY 3rd Annual Meeting, 2 0 - 2 1 April 1994, Skagen, Denmark Organizing committee: Henning Beck-Nielsen Ole Hother-Nielsen Laszlo Hegedüs Karine Bech Claus Hagen Peter Laurberg Jens Sandahl Christiansen Erik Fink Eriksen Niels Erik Skakkebask Jannik Hilsted Niels Juel Christensen Lars 0stergaard Kristensen Supplement 1, Vol. 130 Oslo 1994 Contributors Abstract Abstract 1 , 4 . . . . 19 14 . . . . 9, 12 2 . . . . 10 10 1 2 . . . . 16 3 . . . . 11 21 6 3 . . . . 7, 14 . . . . 13 11 . . . . 12 8 . . . . 20 . . . 9 , 1 2 , 1 8 , 2 0 . . . . 17 5, 17 . . . . 1 ,4 . . . . 10 21 . . . . 13 17 . . . . 10 16 . . . . 14 3 17 . . . . 15 20 7 2, 3 . . . . 16 4 . . 9, 12, 18 2 13 5 4 . . . . 16 2. 3 5 . . . . 6 , 1 1 13 . . . . 15 16 9 . . . . 17 19 16 . . . . 18 . . . . 17 19 13 7 19 10 14 . . . 9, 12, 18, 20 20 3 18 20 6 16 21 11 20 7 15 8 8 EHR I KNDOCRINOL 1 9 9 4 ; 1 3 0 . SUPPL 2 P4.017 P4.019 Poster presentations 237 P4.018 P4.020 I S O L A T I O N A N D I N I T I A L C H A R A C T E R I Z A T I O N O F MICROVASCULAR E N D O T H E L I A L CELLS DERIVED FROM T H E R E T R O O R B I T A L C O N N E C T I V E TISSUE O F PATIENTS WITH GRAVES' O P H T H A L M O P A T H Y . A.Ii. Heufelder. P .C. Scriba, and R.S. Balm*. Molecular Thyroid Research Group, Med. Klinik. Klinikum Innenstadl, München, Germany, and Division of Endocrinology*. Mayo Clinic, Rochester/MN, USA. Activation of certain adhesion molecules in vascular endothelial cells and in the surrounding cxtravascular tissue likely plays an central role in the site-specific recrui tment of immunocompetent cells in Graves' ophthalmopathy (GO). Transendothelial migration of lymphocytes into the cxtravascular reiroorbital space depends upon the expression of a finely tuned cascade of cytokine-act ivatcd adhesion receptors by endothel ia l cells of the retroorbital microvasculature. Systematic investigation of the cellular mechanisms involved in these interactions requires carefully contro l led in vitro experiments , using purified populations of the different cell types involved. To characterize in detail the in teract ions be tween orbi ta l endothel ia l cells (OEC) and immunocompetent cells in vitro, we designed a procedure that allows isolation of microvascular endothelial cells from small volumes of orbital connective tissue. Of the various techniques tested, an indirect approach using successive rounds of incubation with anti-Ulex europaeus I (UEA I) lectin and anti-human endothelial cell (CD31) monoclonal antibody coupled to immunomagnetic polystyrene beads resulted in the recovery of viable O E C with high yield and purity (>99%). Following isolation of OEC, roseited magnetic beads were detached using excess fucosc (0.01 M). Endothelial origin of the resulting cell population was demonstrated by the characteristic cobblestone-like appearance of OEC monolayers, and by specific positive immunorcactivity when using antibodies directed against von Willebrand factor, thrombomodulin, and human endothelial cells (CD31). Following exposure to tumor necrosis fac tor-a( l00 U/ml). immunoperoxidase staining of primary cell monolayers revealed positive immunoreactivity for endothel ial cell-specific adhesion molecule-1 (ELAM- l ) and vascular cell adhesion molecule (VCAM-l ) . Negative immunos ta in ing us ing m o n o c l o n a l ant ibodies directed against cytokcrat in, vimentin and fibroblast antigen further confirmed the homogenei ty of the O E C populat ion obtained. In conclusion, this technique allows highly purified OEC to be isolated from small volumes of orbital connect ive t issue with high yield and reproducibili ty. Availability of OEC provides an important tool for the analysis of the cellular mechanisms involved in the site-directed recruitment of lymphocytes in GO. Further, experiments using OEC in vitro may help to explore the potential benefits of therapy specifically designed to block lymphocyte access to the retroorbital space at the level of the endothelial cell. Supported by DFG (He 1485/3-1) 

Isolation and initial characterization of microvascular endothelial cells derived from the retoorbital connective tissue of patients with Graves ophthalmopathy

Universität München: Elektronischen Publikationen

http://epub.ub.uni-muenchen.de/8867/1/8867.pdf

Isolation and initial characterization of microvascular endothelial cells derived from the retoorbital connective tissue of patients with Graves ophthalmopathy

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