TSH-R expression and cytokine profile in orbital tissue of active vs. inactive Graves' ophthalmopathy patients.
OBJECTIVE
From in vitro studies using cultures of orbital fibroblasts, it has become clear that cytokines play an important role in the orbital inflammation in Graves' ophthalmopathy (GO). Orbital fibroblasts seem to be the key target cells of the autoimmune attack, and they are able to express the TSH receptor (TSH-R). In vivo data on the presence of cytokines in orbital tissues are sparse, and mostly limited to samples obtained from patients with endstage, inactive GO; the same holds true for the presence of the TSH-R. The aim of the present study was to determine whether the cytokine profile and TSH-R expression differ in the active vs. the inactive stage of GO.


METHODS
Orbital fat/connective tissue was obtained from six patients with active, untreated GO undergoing emergency orbital decompression, and from 11 patients with inactive GO subjected to rehabilitative decompressive surgery. The mRNA levels of various cytokines and the TSH-R were assessed by real-time polymerase chain reaction (PCR) using the LightCycler. Data are expressed as ratios (unknown mRNA/beta-actin mRNA).


RESULTS
Active GO patients had much higher TSH-R expression than inactive patients: 4/0-24 (median value/range) vs. 0/0-9, P = 0.01. TSH-R expression was related to the Clinical Activity Score (r = 0.595, P = 0.015). Patients with active GO compared to those with inactive GO had higher mRNA levels of the proinflammatory cytokines interleukin-1beta (IL-1beta) (445/153-877 vs. 0/0-455, P = 0.001), IL-6 (1583/968-18825 vs. 559/0-7181, P = 0.01), IL-8 (1422/38-7579 vs. 32/0-1081, P = 0.046) and IL-10 (145/58-318 vs. 27/0-189, P = 0.002). In active GO there also existed a trend towards a predominance of T helper 1 (Th1)-derived cytokines as evident from higher IL-2 (37/0-158 vs. 0/0-68, P = 0.043), interferon-gamma (IFN-gamma) (20/0-79 vs. 0/0-16, P = 0.12) and IL-12 (2.3/0-14.8 vs. 0/0-1.6, P = 0.10) mRNAs. IL-1 receptor agonist (IL-1RA), IL-2 receptor (IL-2R), IL-3, IL-4, IL-5, IL-13, IL-18 and tumour necrosis factor-alpha (TNF-alpha) mRNAs were similar in both groups.


CONCLUSIONS
These data show that at the mRNA level, TSH-R expression is largely present only during the active stages of GO. The active phase is characterized by the presence of proinflammatory and Th1-derived cytokines, whereas other cytokines, among them Th2-derived cytokines, do not seem to be linked to a specific stage of GO.
Wakelkamp
I M M J
IM
Department of Endocrinology, Academic Medical Centre, University of Amsterdam, Amsterdam, the Netherlands. I.M.M.J.Wakelkamp@amc.uva.nl
Bakker
O
O
Baldeschi
L
L
Wiersinga
W M
WM
Prummel
M F
MF
D000208:Acute Disease; D017667:Adipocytes / Q000378:metabolism*; D000328:Adult; D003239:Connective Tissue Cells / Q000378:metabolism*; D016207:Cytokines / Q000378:metabolism*; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D006111:Graves Disease / Q000378:metabolism* / Q000601:surgery; D006801:Humans; D007371:Interferon-gamma / Q000235:genetics; D007378:Interleukins / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D009915:Orbit; D012333:RNA, Messenger / Q000032:analysis; D011989:Receptors, Thyrotropin / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018709:Statistics, Nonparametric
D016207:Cytokines; D007378:Interleukins; D012333:RNA, Messenger; D011989:Receptors, Thyrotropin; D007371:Interferon-gamma