Innate Immunity, Microbiology, Inflammation | ABSTRACTS
1006
Type 2 immune cells selectively interact with skin sensory nerve fibers in
atopic dermatitis
L Oetjen, L Yang, T Whelan, S Hamilton, P Wang, M Miller and B Kim Washington University
School of Medicine, St. Louis, MO
Chronic itch is frequently cited as the most debilitating symptom of atopic dermatitis (AD).
Although the type 2 cytokine IL-4 has recently been shown to be a critical molecular mediator
of chronic itch, the cellular interactions between type 2 immune cells and sensory neurons in
AD are not well understood. To explore this neuroimmune interface, we generated a dual
fluorescent reporter mouse line in which sensory neurons express tdTomato and immune cells
express IL-4-specific enhanced GFP (eGFP). Using intravital two-photon imaging of skin le-
sions in a mouse model of AD, we observed that a subset of IL-4-eGFP+ type 2 immune cells
markedly reduced their instantaneous speed upon interacting with sensory nerve fibers but
resumed rapid transit after leaving the fibers. Overall, IL-4-eGFP+ immune cells that associ-
ated with sensory nerve fibers had a lower mean speed over the course of imaging compared
to IL-4-eGFP+ cells that did not make contact with sensory fibers. In contrast, type 1 and type
3 immune cells did not demonstrate similar neuroimmune interactions. Collectively, we have
identified a selective behavior of type 2 immune cells towards sensory nerve fibers in the skin.
These observations provide a novel platform to identify key mediators of neuroimmune in-
teractions that underlie AD.
1008
Type I immunity induces a senescence-like growth arrest in malaria parasites
D Sossau1, D Mazier2 and M Rocken1 1 Department of Dermatology, University Hospital
Tubingen, Tubingen, Germany and 2 UPMC UMRS CR7, Paris, Ile-de-France, France
Type I immunity kills and contains plasmodium species in mammalians. However, malaria
tends to recur, showing that the natural type I immune response strongly reduces the path-
ogens but fails to eradicate them. Analyzing the effect of type I immunity on plasmodium-
infected hepatocytes, we confirmed that indeed IFN (interferon)-ã strongly reduces parasite
number. Surprisingly, we also found that IFN-g induces in addition a senescence-like growth
arrest in the remaining plasmodium parasites. One day of treatment with IFN-g caused a
stable growth-arrest over 14 days in surviving Plasmodium (P.) falciparum liver stages. IFN-g
induced small non-proliferating plasmodium forms in human, monkey and mouse hepato-
cytes. Atovaquone treatment killed the large majority of the remaining parasites confirming
that these small forms are alive. In vitro data showed that mammalian p21 can interact with
parasite cyclin-dependent kinases (CDK) PFMRK, PFPK5 and PFPK6. Inhibition of these CDKs
with Artemisinin severely impairs the growth of P.falciparum, without killing the parasites.
Therefore, we asked whether IFN-g and plasmodium parasites induce p21 in mammalian
hepatocytes and whether mammalian p21 can arrest P.falciparum in vivo. Liver-infection
with plasmodium parasites leads to an interferon-response and induced p21 but not p16
protein levels inside primary hepatocytes. The p21 protein was further enhanced by treatment
with IFN-g and tumor necrosis factor. Most importantly, p210/0 hepatocytes had a major
defect in containing malaria parasites in vitro. In consequence, infection of p210/0 mice had a
major defect in controlling the onset and the extent parasitemia. Hence, type I immunity did
not only kill malaria parasites but induced a p21-dependent senescence-like growth arrest in
the remaining plasmodium stages. This IFN-g-induced senescence-like growth arrest signif-
icantly contributed to the containment of the disease.
1010
Nummular eczema is a distinct clinical entity with overlapping features of
both, psoriasis and atopic eczema
A Boehner1, F Lauffer2, T Biedermann2 and K Eyerich3 1 Department of Dermatology and
Allergy, Technical University of Munich, Munich, Germany, 2 1, Munich, Germany and 3
Department of Dermatology and Allergy, Technische Universität Munich, Munich, Germany
Background: Nummular eczema (NE) is a chronic inflammatory skin disease that is charac-
terized by multiple, pruritic, discoid-shaped eczematous lesions. Objectives: We sought to
better understand the disease characteristics of NE and compare it to Atopic eczema (AE) and
psoriasis. Methods: 31 patients with NE, 40 patients with AE and 65 patients with psoriasis
confirmed by clinical and histological evaluations were included in this study. We compared
clinical, histopathological and immunohistochemical as well as RNA sequencing patterns
between these groups. Results: The atopic characteristics, serum IgE levels and blood eo-
sinophils were highly significantly elevated in AE (median total IgE: 627 kU/L; eosinophilia:
71%) compared to NE (105 kU/L; 32%), while there was no significant difference between NE
and psoriasis (67,4 kU/L; 19%), although both diseases presented higher IgE levels compared
to healthy controls (27,9 kU/L). As expected, colonization with Staphylococcus aureus (SA)
on lesional skin was most pronounced in AE (85%). However, NE (52%) was significantly
more often colonized by SA than psoriasis (17%). Histologically, lesional epidermis showed
greater acanthosis in NE (322 mm) than in AE (230 mm), but was less pronounced compared to
psoriasis (484 mm). Significant intralesional neutrophilic infiltration was more often present in
NE (52%) compared to AE (8%), while there was no difference considering intralesional
eosinophils between AE and NE. Consistent with the clinical and histological data, immu-
nohistochemistry revealed a higher expression of neutrophil elastase (14 vs. 2 cells/HPF) and
Ki67 (188 vs. 129 cells/HPF) and a lower expression of the Fcå-recepter (106 vs. 52 cells/HPF)
in NE compared to AE. Furthermore, RNA sequencing revealed a significant upregulation of
the IL-17 pathway in NE, but not in AE. Conclusion: Clinical and histological evidence
suggest that NE is a distinct clinical entity rather than a variant of AE with overlapping features
of both AE and psoriasis.
1007
Analysis of cutaneous microbiota between two age-group of Caucasian women
P Rouaud-Tinguely1, R Jugé1, C Mainzer2, D Boudier3, M Roth4, H Coppin4 and B Closs1 1
SILAB R&D Department, St. Viance, Limousin, France, 2 SILAB Inc., Hazlet, NJ, 3 SILAB
R&D Department, Saint Viance, Limousin, France and 4 IRSD, Université de Toulouse,
INSERM, INRA, ENUT, Toulouse, France, Toulouse, Midi-Pyrenees, France
Although skin aging was deeply investigated, no study has identified potential age-related
changes in cutaneous microbiota for Caucasian ethnicity. In this context, the aim of our study
was to compare the microbiota diversity between two different age groups of Caucasian
women. For this purpose, skin-swab samples were collected directly on the forehead of 34
healthy Caucasian women: 17 younger (aged 2131 years) and 17 older individuals (aged
5469 years). Bacterial communities were evaluated using the 16S rRNA gene sequencing.
Data revealed a significant higher Shannon alphadiversity index in older (1.74
0.35) than in
younger skin (1.30
0.44), indicating an impact of aging on the evenness but not in the
richness of the skin microbiota. Overall microbiota structure was different between the two
age groups, as demonstrated by beta-diversity analysis. Furthermore, taxonomic composition
analysis showed both an increase of Proteobacteria and a decrease of Actinobacteria in older
skin. At genus level, older skin exhibited a significant increase of Corynebacterium (+84%)
and a decrease of Propionibacterium (-40%) relative abundance. The modifications in pH
level, sebum secretion or immune defenses that are characteristic of aged skin lead to a lower
selective pressure of commensal bacteria, so that opportunistic such as Proteobacteria and
Corynebacterium can colonize the skin, leading to a higher diversity. In conclusion, our study
represents the first identification of skin microbiota shift during aging of Caucasian women.
This study will support new approaches in order to rebalance skin microbiota and prevent
age-related skin disorders.
1009
Demodex mites modulate skin inflammation: Potential role in rosacea
S Gatault1, R Foley1, L Shiels1 and F Powell2 1 Charles Institute of Dermatology, University
College Dublin, Ireland and 2 Charles Institute of Dermatology, University College Dublin,
Dublin, Ireland
Low number of Demodex mites reside in the upper pilosebaceous follicles of healthy human
facial skin in contact with keratinocytes without inducing inflammation. In rosacea patients,
the greatly increased follicular mite population and the resulting follicular distension/
disruption exposes them to both sebocytes and PBMCs (Peripheral Blood Mononuclear Cells).
The aim of this study is to evaluate how Demodex mite density modulates inflammation in
keratinocytes, sebocytes and PBMCs. Mites isolated from human skin using the Modified
Standardized Skin Surface Biopsy (MSSSB) were incubated with human keratinocytes
(HaCaT), rat preputial sebocytes (RPSs) and human PBMCs for 24 hours at low (1 mite/cm2)
and high (5 mites/cm2) densities. TLR2 expression and inflammatory mediators release were
assessed by flow cytometry, qRT-PCR, Luminex array and ELISAs. Keratinocytes showed a
higher gene and protein expression of TLR2 when they were incubated with mites. However,
only a high mite density significantly increased the gene expression and release of inflam-
matory mediators (eg. KLK5 (p<0.05) and CCL2 (p<0.05), n¼5). RPSs and PBMCs released
significant levels of chemokines and inflammatory mediators (eg. CCL2 (p<0.05), MIF
(p<0.05), CXCL3 (p< 0.001), TNFa (p<0.01) and IL-6 (p< 0.01), n¼3) after incubation for 24
hours with even a low number of live Demodex mites. The contrasting effects on inflam-
matory pathways described here provide insight into how these complex organisms may co-
exist with host cells under normal skin conditions (exposure to keratinocytes in low numbers)
without disrupting the biobalance, and yet have the capacity to induce significant cellular
inflammatory reactivity when their population is markedly increased (keratinocytes) or when
they are exposed to other cells (sebocytes and PBMCs) as may occur in the skin of patients
with rosacea.
1011
IL-17 is crucial for psoriatic inflammation, but also initiates an anti-
inflammatory feedback loop via signaling into keratinocytes
H Ha, H Wang and U Siebenlist IAS/ LMI/ NIAID/ NIH, Bethesda, MD
The IL-17 cytokines make crucial contributions to chronic inflammatory skin disease and anti-
IL-17 antibodies have proven to be highly efficacious in treating psoriasis. IL-17 cytokines
signal via the obligatory adaptor CIKS/Act1. Previously, we have identified critical roles of
CIKS-mediated signaling in imiquimod-induced psoriatic inflammation. IL-17/CIKS-mediated
signaling into keratinocytes was essential for neutrophil infiltration and contributed to pa-
thology by promoting proliferation and dampening terminal differentiation of keratinocytes.
Interestingly, IL-17 signaling into keratinocytes also exerted negative feedback, delimiting
continued accumulation of IL-17-producing gd T cells in skin. We now demonstrate that the
negative feedback is mediated in part via IL-19, a member of the IL-10 family of cytokines. IL-
19 is strongly induced by IL-17-stimulated keratinocytes and appears to function by sup-
pressing recruitment of IL-17-producing gd T cells over time, thereby dampening IL-17
production. This stands in contrast to IL-17/CIKS-mediated signaling into non-keratinocytes -
specifically dermal fibroblasts - which initially promoted cellular infiltration and enhanced
accumulation of IL-17-producing gd T cells in skin, comprising an early positive feed-forward
mechanism. Our data show that CIKS-mediated signaling is dynamic; it is central to the
development of both dermal and epidermal hallmarks of psoriasis, but also initiates mech-
anisms that ultimately mitigate further tissue damage. The latter mechanisms may offer
additional avenues for therapeutic intervention.
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