The role of phagocytic NADPH oxidase and innate immune cells in the immune activation in toluene diisocyanate (TDI)-induced lung inflammation

0 1 2 3 4 5 To tal NCR + ILC3 (x10 4 ce lls )

SOL TDI SOL TDI 0

10 20 30 40 50 * To tal ex-ILC3 (x10 4 ce lls )

SOL TDI SOL TDI * 0 5 10 15 20 25 30

*

SOL TDI SOL TDI

*

SOL TDI SOL TDI

ILC1 ILC2 Total ILC3

NCR+ILC3 NCR-ILC3 ex ILC3

0 50 100 150 200 250 300 350 400 450 500 IL -18 (p g/ mg p rotein)

SOL TDI SOL TDI 0

50 100 150 200 250 300 350 400 450 500

*

SOL TDI SOL TDI

*

SOL TDI SOL TDI

IL-18 IL-33 IL-1 IL-17A

The role of phagocytic NADPH oxidase and innate immune cells in the immune

activation in toluene diisocyanate (TDI)-induced lung inflammation

Innate lymphoid cells (ILCs) act as an orchestrator of immunity. They respond to

epithelium derived signals by expressing cytokines and cell surface receptors to mediate

immune regulation. In recent researches, group 2 ILCs (ILC2s) were found to mediate

allergic lung inflammation. ILC2s can be activated by epithelial cell-derived cytokines IL-33,

25 or TSLP, and release mediators associated with a Th2 immune response including

IL-5, IL-9 and IL-13 in allergic lung inflammation. Isocyanates are low-molecular-weight

compounds noted for inducing occupational and environmental asthma.

Isocyanate-induced lung disease, an oxidant stress-dependent pulmonary inflammation, is the leading

cause of occupational asthma. Our previous studies indicated that phagocytic NADPH

oxidase is an essential regulator in TDI-induced airway inflammation characterized by a

server lung inflammation with both Th17 and Th2 immune components.

• Innate lymphoid cells (ILCs) activation may play an important role in phagocytic NADPH

oxidase mediated downstream immune mechanism in TDI-induced airway inflammation.

Figure1. TDI exposure induced airway allergic inflammatory phenotypes in wild

type (WT) but not in Ncf1

mice.

Chia-Ying Cheng

, Chi-Chang Shieh

1

_{Department of Microbiology & Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan}

Figure4. IL-1



IL-23 stimulation induced change of exILC3 phenotype to NCR- ILC3 which

produce more IL-17A.

Figure3. The numbers of ILC2 and NCR-ILC3 significantly increased in lung tissue after

TDI exposure in WT but not in Ncf1

mice.

Figure2. TDI exposure induced more IL-1



and IL-17A production in lung inflammation

of WT when compared with Ncf1

mice.

Introduction

Hypothesis

Results

Conclusion

Purified NKp46-_{ILC3 (NCR}-_{ILC3), NKp46}+_{ILC3 (exILC3), and NKp46}+_CCR6+ _(NCR+ _{ILC3) from lung tissue were cultured in}

RPMI medium with IL-2 (50 ng/ml)and IL-7 (10 ng/ml), and then cultured for 4 days with IL-1 (50 ng/ml) and IL-23 (50

ng/ml) condition medium. (A)Cells were phenotyped for expression of CCR6 and NKp46. (B)The percentage of ILC3 subsets were normalized with total ILC3 cells.

The severity of allergic lung inflammation was measured on days 0- 19 with body weight (A) and airway resistance (B)

after TDI exposure WT and Ncf1-/- mice. (WT, n=8; Ncf1-/-, n=6; Results are shown as mean  SEM. **P  0.01, ***P

 0.001.) The characteristic histopathological images of HE stained (C) and Tri-chrome stained (D) lung tissue from

wild-type and Ncf1-/- _{mice with TDI-induced lung inflammation. The reference scale bars stand for 0.2 mm. Original}

magnification ×100. The mice lung histology photographs represent 1 of 4 mice in each group.

(A)

(B)

(C)

(D)

The expression of inflammatory cytokines in lung tissue homogenates after TDI-challenged WT and Ncf1–/– mice were

detected by ELISA measurement. (WT, n=6; Ncf1-/-, n=4; Results are shown as mean  SEM, *P < 0.05.)

Lung ILC cells in the setting of AAI exacerbation caused by TDI stimulation. A population of lineage negative, CD45.2

positive has in identify in the inflammation of lung tissue of WT and Ncf1-/- mice. Data are from 2 independent experiments

(n= 3 mice per group) expressed as mean  SEM, *P < .05.

(A)

_(B)