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Understanding chemical allergen potency: role of NLRP12 and Blimp-1 in the induction of IL-18 in human keratinocytes

  • Immunotoxicology
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Abstract

Keratinocytes (KCs) play a key role in all phases of skin sensitization. We recently identified interleukin-18 (IL-18) production as useful end point for determination of contact sensitization potential of low molecular weight chemicals. The aim of this study was to identify genes involved in skin sensitizer-induced inflammasome activation and to establish their role in IL-18 production. For gene expression analysis, cells were treated for 6 h with p-phenylenediamine (PPD) as reference contact allergen; total RNA was extracted and examined with a commercially available Inflammasome Polymerase Chain Reaction (PCR) array. Among genes induced, NLRP12 (Nod-like receptor P12) was selected for further investigation. NLRP12 promoter region contains Blimp-1 (B-lymphocyte-induced maturation protein-1)/PRDM1 binding site, and from the literature, it is reported that Blimp-1 reduces NLRP12 activity and expression in monocytes/macrophages. Their expression and role in KCs are currently unknown. To confirm NLRP12 expression and to investigate its relationship with Blimp-1, cells were exposed for different times (3, 6 and 24 h) to the extreme sensitizer 2,4-dinitrochlorobenzene (DNCB) and the strong sensitizer PPD. Allergens were able to induce both genes, however, with different kinetic, with DNCB more rapidly upregulating Blimp-1 and inducing IL-18 production, compared to PPD. NLRP12 and Blimp-1 expression appeared to be inversely correlated: Blimp-1 silencing resulted in increased NLRP12 expression and reduced contact allergen-induced IL-18 production. Overall results indicate that contact allergens of different potency differently modulate Blimp-1/NLRP12 expression, with strong allergen more rapidly downregulating NLRP12, thus more rapidly inducing IL-18 production. Data confirm that also in KCs, NLRP12 has an inhibitory effect on inflammasome activation assessed by IL-18 maturation.

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Abbreviations

ACD:

Allergic contact dermatitis

AOO:

Acetone olive oil

ASC:

Apoptosis-associated speck-like protein

Blimp-1:

B-lymphocyte-induced maturation protein-1

CARD:

Caspase activation and recruitment domain

DAMPs:

Damage-associated molecular patterns

DiSFeB:

Dipartimento di Scienze Farmacologiche e Biomolecolari

DMSO:

Dimethyl sulfoxide

DNCB:

2,4-Dinitrochlorobenzene

ELISA:

Enzyme-linked Immunosorbent assay

IFN-γ:

Interferon-γ

IL-18:

Interleukin-18

IL-1β:

Interleukin-1β

KC:

Keratinocytes

LLNA:

Local lymph node assay

LLR:

Leucine-rich repeat protein

NF-κB:

Nuclear factor-κB

NLR:

NOD-like receptor

NLRP12:

Nod-like receptor P12

PAMPs:

Pathogen-associated molecular patterns

PBS:

Phosphate-buffered saline

PCR:

Polymerase chain reaction

PPD:

p-Phenylenediamine

PVDF:

Polyvinylidene difluoride

PYD:

Pyrin domain

REACH:

Registration evaluation authorization and restriction of chemicals

RHE:

Reconstituted human epidermis

SDS:

Sodium dodecyl sulfate

siRNA:

Small interference RNA

TLRs:

Toll-like receptors

TNF-α:

Tumor necrosis factor-α

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Acknowledgments

This research was partially supported by the Alternatives Research & Development Foundation, 2014—Alternatives Research Grant Program.

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Authors and Affiliations

  1. University of Milan, Milan, Italy

    Angela Papale, Elena Kummer, Valentina Galbiati, Marina Marinovich, Corrado L. Galli & Emanuela Corsini

Authors
  1. Angela Papale
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  2. Elena Kummer
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  3. Valentina Galbiati
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  4. Marina Marinovich
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  5. Corrado L. Galli
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  6. Emanuela Corsini
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Correspondence to Angela Papale.

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Papale, A., Kummer, E., Galbiati, V. et al. Understanding chemical allergen potency: role of NLRP12 and Blimp-1 in the induction of IL-18 in human keratinocytes. Arch Toxicol 91, 1783–1794 (2017). https://doi.org/10.1007/s00204-016-1806-8

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  • DOI: https://doi.org/10.1007/s00204-016-1806-8

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