The Expression of Toll-like Receptors in Idiopathic Inflammatory Myopathies

Geun-Tae Kim; Jun-Hee Lee; Seung-Hoon Baek; Joung-Wook Lee; Mi-Ra Cho; Ju-In Kim; Sun-Hee Lee; Dae-Seong Kim; Sung-Il Kim

doi:10.4078/jkra.2007.14.2.118

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Kim, Lee, Baek, Lee, Cho, Kim, Lee, Kim, and Kim: The Expression of Toll-like Receptors in Idiopathic Inflammatory Myopathies

Original Article

J Korean Rheum Assoc 2007;14(2):118-124.

Published online: 23 June 2007

DOI: https://doi.org/10.4078/jkra.2007.14.2.118

The Expression of Toll-like Receptors in Idiopathic Inflammatory Myopathies

Geun-Tae Kim^∗∗∗, Jun-Hee Lee, Seung-Hoon Baek, Joung-Wook Lee, Mi-Ra Cho, Ju-In Kim^∗∗, Sun-Hee Lee, Dae-Seong Kim^∗, Sung-Il Kim

^∗Departments of Internal Medicine and Neurology, Busan, Korea

^∗∗Research Institute, Busan, Korea

College of Medicine, Pusan National University, Busan, Korea

^∗∗∗Department of Internal Medicine, Pusan Medical Center, Busan, Korea

Received 24 January 200719 March 2007

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Methods:

The expression of TLR-2, 4 and 9 was measured by real-time RT-PCR and immunohistochemical stain (IHS) from muscle tissues in patients with IIMs and controls.

Results:

The expression levels of TLR-2, 4 and 9 in IIMs were significantly higher than controls. TLR-2, 4 and 9 were mainly expressed on sarcolemma of muscle fibers, perimysial vascular endothelium and infiltrating inflammatory cells in dermatomyositis, whereas, they were mainly expressed on sarcolemma of muscle fibers, destructed muscle fibers, and enodmysial infiltrating inflammatory cells in polymyositis.

Conclusion:

TLR-2, 4 and 9 were highly expressed in muscle tissue of IIMs. These results suggest that TLR-2, 4 and 9 play a role in pathogenesis of IIMs.

Keywords: Toll-like receptors, Idiopathic inflammatory myopathy

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Fig. 1.

Immunohistochemical stain of TLR-2 in muscle tissues from healthy control (A) & patients with IIMs (B, C). (B) In DM, TLR-2 is expressed by sarcolemma of muscle fibers, perimysial vascular endothelium and infiltrating inflammatory cells (arrow). (C) In PM, TLR-2 is expressed by sarcolemma of muscle fibers, destructed muscle fibers, and enodmysial infiltrating inflammatory cells (arrow). Magnification: ∗200.

Fig. 2.

Immunohistochemical detection of TLR-4 in muscle tissues from healthy control (A) & patients with IIMs (B, C). (B) In DM, TLR-4 is expressed by sarcolemma of muscle fibers, perimysial vascular endothelium and infiltrating inflammatory cells (arrow). (C) In PM, TLR-4 is expressed by sarcolemma of muscle fibers, destructed muscle fibers, and enodmysial infiltrating inflammatory cells (arrow). Magnification: ∗200.

Fig. 3.

Immunohistochemical detection of TLR-9 in muscle tissues from healthy control (A) & patients with IIMs (B, C). (B) In DM, TLR-9 is expressed by sarcolemma of muscle fibers, perimysial vascular endothelium and infiltrating inflammatory cells (arrow). (C) In PM, TLR-9 is expressed by sarcolemma of muscle fibers, destructed muscle fibers, and enodmysial infiltrating inflammatory cells (arrow). Magnification: ∗200.

Table 1.

Primers used for real-time RT-PCR

Primer	Stream	Sequence
β-actin	Sense	5 ‘-AACACCCCAGCCATGTACG-3 ‘
	Anti-sense	5 ‘ -ATGTCACGCACGATTTCCC-3 ‘
TLR-2	Sense	5’-CAATGATGCTGCCATTCTCAT-3’
	Anti-sense	5 ‘ -ATTATCTTCCGCAGCTTGCA-3’
TLR-4	Sense	5’-AGTTTCCTGCAATGGATCAAGG-3’
	Anti-sense	5’-CTGCTTATCTGAAGGTGTTGCAC-3’
TLR-9	Sense	5’-AGTCAATGGCTCCCAGTTCCT-3’
	Anti-sense	5’-CGTGAATGAGTGCTCGTGGTA-3’
TNF-α	Sense	5’-ACATACTGACCCACGGCTTC-3’
	Anti-sense	5’-GCACTCACCTCTTCCCTCTG-3’
INF- γ	Sense	5 ‘ -TCCCATGGGTTGTGTGTTTA-3’
	Anti-sense	5 ‘ -AAGCACCAGGCATGAAATCT-3 ‘

Table 2.

Characteristics of 14 patients with IIMs and the expression levels of TLR-2, 4, 9 and TNF-α, IFN-γ

Patient	Sex	Age	Diagnosis	Relative density of mRNA expression compared to controls
Patient	Sex	Age	Diagnosis	TLR-2/β -actin	TLR4/β -actin	TLR-9/β -actin	TNF-α /β -actin	IFN- γ/β -actin
1	F	33	PM	16	48	11	10	587
2	F	59	PM	19	140	15	27	1,431
3	F	30	DM	25	164	6	4	278
4	M	55	DM	15	106	12	14	484
5	M	37	DM	48	61	5	5	273
6	M	22	PM	19	169	15	31	674
7	F	47	PM	16	149	10	8	382
8	M	58	DM	57	81	6	22	463
9	M	37	DM	13	153	37	17	885
10	M	39	DM	70	232	26	20	2,756
11	F	43	PM	50	228	35	24	1,825
12	F	8	DM	40	444	44	51	2,179
13	M	36	DM	36	108	5	3	334
14	F	66	DM	29	297	35	83	3,141
				32±18	170±105	19± 14	23±22	121±978

F: female, M: male, DM: dermatomyositis, PM: polymyositis, IFN- γ : interferon- γ, TNF- α : tumor necrosis factor- α, TLR: toll-like receptor

Table 3.

The expression of TLR-2, 4, 9 and TNF- α, IFN- γ according to disease (PM, DM) and sex (male, female)

	Disease		Sex
	PM (n=5)	DM (n=9)	Male(n=7)	Female (n=7)
TLR-2/β-actin	24±15	37±19	37±22	28±13
TLR-4/β-actin	147±65	183±123	130±59	210±129
TLR-9/β-actin	17±10	20±16	15±12	22±15
TNF- α/γ-actin	20±10	24±26	16±10	30±28
IFN- γ/β-actin	980±617	1,199±1,160	839±870	1,403 ±1,063

F: female, M: male, DM: dermatomyositis, PM: polymyositis, IFN- γ : interferon- γ, TNF- α : tumor necrosis factor- α, TLR: toll-like receptor

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