YAKUGAKU ZASSHI 133(12) 14011409 (2013) 2013 The Pharmaceutical Society of Japan

1401

Review

toll-like receptor-2 (TLR2

Regulation of Expression, Function, and Inammatory Responses of Innate Immune Receptor

Toll-like Receptor-2 (TLR2) during Inammatory Responses against Infection
Tsuyoshi Shuto
Department of Molecular Medicine, Graduate School of Pharmaceutical Sciences, Kumamoto
University; 51 Oe-Honmachi, Chuo-ku, Kumamoto 8620973, Japan.
(Received July 23, 2013)
Toll-like receptor-2 (TLR2) is one of the important innate immune receptors that play an important role in recognizing the pathogens and producing inammatory cytokines. In this review, we focus on the regulatory mechanisms of
expression, function and inammatory responses of TLR2 during pathogenic infection in innate immune cells. We rst
showed that nontypeable Haemophilus inuenzae (NTHi), an important human pathogen that exacerbates otitis media
and chronic obstructive pulmonary diseases (COPD), induces inammatory responses through activation of NF-kB via
two distinct signals, NIK-IKKa/b-IkBa and MKK3/6-p38 pathways. Moreover, TLR2 was greatly up-regulated by
NTHi through the positive IKK-IkBa-dependent NF-kB pathway and the negative MKK3/6-p38a/b pathway. Importantly, glucocorticoids synergistically enhance NTHi-induced TLR2 up-regulation likely via a negative cross-talk
with the inhibitory p38 MAPK. The results provide novel insights into the role of glucocorticoids in regulating host
defense and innate immune responses through TLR2 regulation. We next focused on the pathogenesis of cystic brosis
(CF), a common lethal inherited disorder characterized by recurrent pulmonary infections and obstruction caused by
chronic mucus hypersecretion and inammation. We demonstrated an increased expression of TLR2, due to an enhanced DNA de-methylation and Sp1-dependent transcriptional activation in CF epithelial cells. Furthermore, a Th17
cytokine IL-17A synergistically increased TLR2 signal through an enhancement of p38 phosphorylation. These studies
suggest the importance of TLR2 and IL-17 signals in the pathogenesis of CF. Finally, by focusing on neutrophils and CF
airway epithelial cells, we identied curcumin as a potent inhibitor of TLR2-mediated inammatory responses.
Key wordstoll-like receptor-2 (TLR2); epithelial cell; cystic brosis (CF); curcumin; neutrophil; nontypeable Haemophilus inuenzae (NTHi)

1.

NK

2)

1)

The author declares no conict of interest.

8620973 51
e-mail: tshutogpo.kumamoto-u.ac.jp
24

2011

3)

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90

9) NTHi

NTHi

pattern recognition receptors; PRRs

NTHi

NTHi

PRRs toll-like receptorsTLRs

NTHi

nuclear factor-kB NF-kB )10)

4)

TLRs

NTHi NF-

kB NTHi

TLRs

5 NF-kB

70

5)

NF-kB-inducing kinase (NIK)-IkB kinase

NTHi

TLRs

IKKa/b inhibitor of NF-kBIkBa

TLR26)

MAP

kinase

kinase

(MKK) 3/6-p38 mitogen activated protein kinase

MAPK 2

2.

NF-kB

NTHi

TGF-b-activated kinase 1 (TAK1)11)

IkBa p38 MAPK

NTHi NF-kB

NTHi NF-

kB

TLR2 NTHi NF-kB

NTHi NF-kB

NTHi

P612) NF-kB

chronic obstructive pulmonary disease;

COPD

nontypeable Hae7,8) NTHi

mophilus inuenzae; NTHi

Hi

99 01

06
10
13 99 00
04
05 CPMCRI

No. 12

1403

NTHi NF-kB

cafeic acid phenethyl ester (CAPE), IkBa

13)

MG-132 TLR2

NTHi TLR2-TAK1

reverse transcriptase-polymerase chain reactionRT-

IkBa p38 MAPK

PCR

NF-kB

TLR2

NF-kB p65

3.

NTHi TLR2

TLR2
IkBa IKKb

TLR2

NTHi IKKb-IkBa

TLRs

NF-kB NTHi TLR2

TLR2

p38 MAPK TLR2

6)

TLR2

p38 MAPK SB203580

TLR2 RT-PCR

SB203580 NTHi

TLR2

TLR2 2 5

p38 MAP

NTHi

MKK3 / 6

TLR2

NTHi MKK3 / 6-p38

MAPK NTHi TLR2

TLR2 NTHi 3

4 5 10

Fig. 1, Left
14)

NTHi TLR2

NTHi IKKb-IkBa

14)

NF-kB TLR2

NF-kB TLR2

NTHi

NF-kB

p38 MAPK TLR2

Fig. 1. Schematic Representation of the Signaling Pathway Involved in Dexamethasone (DEX)-mediated Enhancement of NTHi-induced TLR2 Expression via Inhibition of p38 MAPK in Human Epithelial Cells
NTHi up-regulates TLR2 expression via a positive IKK-IkBa-dependent NF-kB pathway and a likely negative MKK3/6-p38a/b pathway. Glucocorticoids, well
known potent anti-inammatory agents, synergistically enhance NTHi-induced TLR2 up-regulation likely via a negative cross-talk with the inhibitory p38 MAPK
pathway. The up-regulated TLR2 leads to enhanced immune and inammatory responses.

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Fig. 1,

Fig. 1

Left NTHi TLR2

TLR2

NF-kB

p38 MAPK NTHi

TLR2

TLR2

4.

NTHi

TLR2
TLR2

NTHi

NTHi TLR2

5.

CF TLR2

TLR2 NF-kB

p38 MAPK

cystic fibrosis; CF CF

NF-kB

CF transmembrane conductance

dexametha-

regulaterCFTRcAMP Cl

sone DEX )15) NTHi TLR2

16) CF

30

NF-kB p38 MAPK

16)

DEX SB203580

CF

NTHi TLR2

2 5 DEX

NTHi TLR2

p38 MAPK

CF TLR2

TLR2

CF

DEX NTHi

17,18) CF

TLR2

TLR2

GR

CF

DEX/GR p38 MAPK

TLR2

CF TLR2

14)
Fig. 1, Right

NTHi

TLR2

16HBE14o-

TLR2

CF CFBE41o- TLR2

TLR2

3.5

TLR2

CFBE41o- TLR2

14)

CF

NTHi

3.5

TLR2

CFTR CF

NTHi TLR2

CFBE41o-/WT-CFTR

CFBE41o- TLR2

No. 12

1405

19)

TLR2

CFTR CLR2

CF

Fig. 2

(B)

DNA 5-azacyti-

CF

dine 5-AC 16HBE14o- TLR2

Fig. 2. A Potential Model Representing the Mechanisms Underlying Hypomethylation-dependent TLR2 Promoter Activation in Human Epithelial Cells
A) Schematic representation showing that the specic CpG sites are highly demethylated in CF epithelial cells and these CpG sites overlap with a minimal region
that is necessary and sucient to maintain basal TLR2 promoter activity. Binding sites for dierent transcription factors are indicated (binding site for: SP1, open
circle; NF-kB, solid circle; ETS, gray circle). Indicated black lines are CpG sites that can be methylated. Transcriptional start site is indicated as 1. Methylation
status of CpGs that are highly demethylated and modestly demethylated in CF cells are indicated as open box and grey box, respectively. Minimal region that is
necessary for basal TLR2 expression is indicated as solid box. B) A model for the mechanisms of DNA demethylation-dependent TLR2 promoter activation in human epithelial cells. As shown in left panel, increased methylation of CpG#18
20 within TLR2 promoter is observed in non-CF epithelial cells, followed by the enhanced recruitment of unknown factors ``X'' that could recognize methylated CpGs and suppress SP1-dependent transcriptional activation without aecting the
binding of SP1 to those CpGs. 5-Azacytidine (5-AC) treatment of non-CF cells leads to inducing demethylation of CpG#1820. On the other hand, CF epithelial
20, which might abolish the recruitment and activity of factors ``X'' in these cells.
cells express high level of TLR2 gene through decreased methylation of CpG#18
The existence of CFTR-dependent epigenetic regulation at specic CpGs in human epithelial cells is intimated by the fact that the introduction of WT-CFTR gene
into CFBE41o- (CF) cells increased the methylation of CpG#18
20.

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DNA

IL-17A

TLR2 bisul-

te sequence 16HBE14o-

CFBE41o- TLR2

IL-17A KO

DNA CFBE41o-

Citrobacter rodentium, Staphylococcus aureus, Kleb-

TLR2

siella pneumoniae, Candida albicans

CF

IL-17A

TLR2 TLR2

DNA

22)

COPD

Fig.

23) IL-17A

19)

2(B)

TLR2

TLR2

IL-17A

TLR2

SP1 SP1 2

CF

TLR2

TLR2 IL-8

Fig. 2 ( A )

IL-17A

SP1 2 SP1

CFBE41o- DHBE-CF

DNA

CF IL-17A

TLR2

IL-17A

Fig.

20)
2(B)

IL-8

CF

IL-8 IL-17A p38 ERK

TLR2

MAPKs

TLR2

IL-17A CFBE41o-

19) DNA

DHBE-CF TLR2

TLR2

IL-8

SP1

IL-17A CFTR

20) CF

16HBE14o-, Calu-3,

A549

6.

CF TLR2

IL-8 IL-17A

NHBEIL-8
CF

IL-17 20 30 kD

IL-17A IL-8 TLR2

IL-17A p38 MAPK

6 IL-17A, IL-17B, IL-

21)
17C, IL-17D, IL-17E, IL-17F

IL-17A p38 MAPK

IL-17A IL-17

CF

Th1 Th2

IL-8

Th17 2005 T

Fig. 3 24) CF

IL-17A

IL-17A

IL-8

p38 MAPK

No. 12

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curcumin
TLR2
PMN
curcumin TLR2
IL-8
curcumin TLR2
Fig. 3. Schematic Representation of the Mechanism Responsible for the Synergism between IL-17A and TLR2 Signal to
Induce IL-8 Expression in CF Airway Epithelial Cells
IL-17A synergistically increases IL-8 production induced by TLR2
agonist peptidoglycan (PGN) in a human CF bronchial epithelial cell line
and primary cells. IL-17A-dependent synergism appears to be the result of
enhanced PGN-induced phosphorylation of p38 MAPK.

N-acetylcysteine NAC
curcumin TLR2
27)
curcumin CF
TLR2
CFBE41o- DHBE-CF

CF

CFBE41o- DHBE-CF

curcumin TLR2

curcumin

mRNA

CF TLR2

CF

curcumin TLR2

SP1 TLR2

CF

7.

TLR2

curcumin

TLR2

27) Curcumin

DMSO

bioavailability

HL-60 dHL-60

polymor-

TLR2

phonuclear leukocyte; PMN

8.

dHL-60 PMN

TLR2

dHL-60 TLR2

PMN

COPD

25)

CF TLR2

dHL60

THP-1 TLR2

14,19) CF IL-17A

24)

TLR2

dHL60, THP-1

25)

curcumin TLR2

curcumin CF TLR2

curcumin 40 mM

TLR2

27)

curcumin

40 mM TLR2

TLR2

curcumin

26)

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11)

COPD CF
28)
TLRs TLRs

12)

13)

14)

Mary Ann Suico

Jian-Dong Li
Dieter Gruenert

15)

16)

17)

18)

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