the benzene metabolite 1 4 benzoquinone reduces t reg function a potential mechanism for tobacco smoke associated atopic dermatitis

The benzene metabolite 1,4-benzoquinone reduces Treg function – a potentialmechanism for tobacco smoke-associated atopic dermatitis Marcus Winter, MSc, Loreen Thürmann, MSc, Zuguang Gu,

The benzene metabolite 1,4-benzoquinone reduces Treg function – a potential

mechanism for tobacco smoke-associated atopic dermatitis

Marcus Winter, MSc, Loreen Thürmann, MSc, Zuguang Gu, PhD, Gerrit Schüürmann,

PhD, Gunda Herberth, PhD, Denise Hinz, PhD, Martin von Bergen, PhD, Hauke

Harms, PhD, Sven Olek, PhD, Stefan Röder, PhD, Michael Borte, MD, Roland Eils,

PhD, Irina Lehmann, PhD, Saskia Trump, PhD

DOI: 10.1016/j.jaci.2017.01.034

Received Date: 7 November 2016

Revised Date: 22 December 2016

Accepted Date: 23 January 2017

Please cite this article as: Winter M, Thürmann L, Gu Z, Schüürmann G, Herberth G, Hinz D, von Bergen M, Harms H, Olek S, Röder S, Borte M, Eils R, Lehmann I, Trump S, The benzene metabolite 1,4-benzoquinone reduces Treg function – a potential mechanism for tobacco smoke-associated atopic

dermatitis, Journal of Allergy and Clinical Immunology (2017), doi: 10.1016/j.jaci.2017.01.034.

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ENSG00000265479.1 0.000 0.000 DTX2P1-UPK3BP1-PMS2P11 processed_transcript

ENSG00000272752.1 0.309 0.433 STAG3L5P-PVRIG2P-PILRB processed_transcript

ENSG00000273333.1 0.000 0.000 XXbac-BPG300A18.13 3prime_overlapping_ncrna

Cytokine Tconv:Treg

16:1 40.6 (26.9-52.9) 55.2 (35.4-171.4)

Granzyme A

4:1 13.1 (8.6-24.8) 11.1 (7.3-33.7)

- 8:1 32.9 (28.3-38.6) 30.0 (26.5-41.8)

16:1 19.7 (17.3-27.7) 27.9 (13.6-41.2)

Granzyme B

4:1 34.6 (21.2-254.6) 283.5 (50.2-607.4)

0.004 8:1 149.2 (57.5-170.4) 292.1 (234.5-386.9)

16:1 191.5 (66.6-265.7) 301.5 (263.5-549.4)

*two-way ANOVA

Granzyme B concentrations in supernatants of CD3/CD28 & IL-2 stimulated and

(A) mothers and (B) children compared to the entire LINA cohort

(A)

Parameters Analysed subcohort Entire LINA cohort P value*

Mothers

n (%), N = 605 n (%), N = 629 Maternal age at birth

* P value from chi squared test for cross relationship, subgroup against total cohort

fever

“Mittlere Reife”; high, 12 yrs of schooling or more “(Fach-)hochschulreife”

* P value from chi squared test for cross relationship, subgroup against total cohort

fever

“Mittlere Reife”; high, 12 yrs of schooling or more “(Fach-)hochschulreife”

was available compared to the entire LINA cohort with respect to the development of atopic dermatitis

Parameters Analysed subcohort Entire LINA cohort P value*

Children

n (%), N = 500 n (%), N = 629 Atopic dermatitis (AD) up to the age of six

OOH

OOH

E,E-muconic acid

via GST

ONHS

OOHC

CYP2E1

OH

OHNQO1

MPOO

O

epoxidehydrolase

OH

OH

CYP

OHOH

O

benzene dihydrodiol benzene diol epoxide

dihydrodioldehydrogenase

OHOH

catechol

hydroquinone 1,4-benzoquinone

NQO1MPO

OO

BD FACS Aria SORP - IZKF Leipzig

in vitro expansion

cryopreservation

- after 2 weeks expansion

- FoxP3-TSDR methylation

- FoxP3 protein expression

in vitro exposure

24 h

- FoxP3-TSDR methylation

- FoxP3 protein expression

- cytokine detection (CBA)

- liquid nitrogen

plate cells/well beads:cell U/ml IL-2

1st 96

4:1

300

2nd 24

1:1 300

% CD25 + FoxP3 + median (IQR) DMSO 95.2 (92.3 – 97.5), n = 5

50 nM 1,4-BQ 94.9 (91.8 – 98.0), n = 5

97.40.82

FoxP3

97.70.58

1.670.06

A

B