Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2015 Nov;135(11):2824-2833.
doi: 10.1038/jid.2015.207. Epub 2015 Jun 8.

Langerhans Cells Facilitate UVB-Induced Epidermal Carcinogenesis

Julia M Lewis  1 , Christina D Bürgler  1 , Marianna Freudzon  1 , Kseniya Golubets  1 , Juliet F Gibson  1 , Renata B Filler  1 , Michael Girardi  2
Affiliations

Langerhans Cells Facilitate UVB-Induced Epidermal Carcinogenesis

Julia M Lewis et al. J Invest Dermatol. 2015 Nov.

Abstract

UVB light is considered the major environmental inducer of human keratinocyte (KC) DNA mutations, including within the tumor-suppressor gene p53, and chronic exposure is associated with cutaneous squamous cell carcinoma formation. Langerhans cells (LCs) comprise a dendritic network within the suprabasilar epidermis, yet the role of LCs in UVB-induced carcinogenesis is largely unknown. Herein we show that LC-intact epidermis develops UVB-induced tumors more readily than LC-deficient epidermis. Although levels of epidermal cyclopyrimidine dimers following acute UVB exposure are equivalent in the presence or absence of LCs, chronic UVB-induced p53 mutant clonal islands expand more readily in association with LCs, which remain largely intact and are preferentially found in proximity to the expanding mutant KC populations. The observed LC facilitation of mutant p53 clonal expansion is completely αβ and γδ T-cell independent and is associated with increased intraepidermal expression of IL-22 and the presence of group 3 innate lymphoid cells. These data demonstrate that LCs have a key role in UVB-induced cutaneous carcinogenesis and suggest that LCs locally stimulate KC proliferation and innate immune cells that provoke tumor outgrowth.

PubMed Disclaimer

Conflict of interest statement

CONFLICT OF INTEREST

The authors state no conflict of interest.

Figures

Figure 1
Figure 1. Langerhans cells influence UVB-induced cutaneous tumor development
Carcinogenesis was induced in hairless (hr/hr) LC-intact (NLC) and LC-deficient (DTA) mice by exposure to 400 J/m2 UVB three times per week. Increased tumor number (a) and volume (b) were seen in the presence of LC (n=10 NLC, 9 DTA, * P<0.05, **P<0.01, ***P<0.001). (c) Representative images from (a, b).
Figure 2
Figure 2. Langerhans cells do not affect UVB-induced CPD formation
(a) Hairless (hr/hr) LC-intact (NLC) and LC-deficient (DTA) mice were treated with a single dose of UVB and epidermal sheets prepared 1 or 24 hr later and stained with antibodies directed against thymine dimers (CPD) and CD207 plus the nuclear dye To-Pro-3 (shown in Supplementary Figure S1 online). CPD fluorescence intensity was similar in LC-intact and LC-deficient mice. (b) Representative images from (a) show CPD (red) and LC (CD207, green). Scale bar = 20 μm. (c) LC density does not change during the 24 hrs following a single dose of UVB. n=3–8 mice/group; in (c) each symbol represents one mouse.
Figure 3
Figure 3. Langerhans cells persist during chronic UVB exposure and facilitate p53 island formation
CD207+ LC and TCRγδ+ DETC were quantified in epidermal sheets prepared from untreated vs chronic UVB treated (400 J/m2, 3x/wk, 5 wks) LC-intact NLC (a) and LC-deficient DTA (b) hr/hr mice. Representative images (c) show CD207 (green) and TCRγδ (red) staining. In both NLC and DTA mice, DETC are nearly eliminated by chronic UVB treatment, however LC in NLC mice are present at 66% of control levels following 5 wks UVB; *P<0.5, ***P<0.001, n=6 mice/group. Mutant KC p53 island density (d) and area (e) were quantified in epidermal sheets prepared from n=6 NLC and DTA hr/hr mice following 5 or 9 wks UVB exposure. Inset graphs show distribution at 9 wks; each symbol represents one mouse, **P<0.01, ***P<0.001. Representative images (f, h) of p53 islands (red) and CD207+ LC (green), scale bar = 20 μm, and a 3D rendering (g) of p53 (green) and LC (blue). CD207+ LC density (i) is increased in association with p53 islands; **P<0.01. Quantitation is described in Supplementary Figure S3 online.
Figure 4
Figure 4. LC facilitation of p53 island growth is T cell independent
Mutant KC p53 island density (a) and area (b) were quantified in epidermal sheets prepared from TCRβ−/−δ−/−.NLC and TCRβ−/−δ−/−.DTA mice following 5 or 9 wks UVB exposure (400 J/m2, 3x/wk). Inset graphs show distribution at 9 wks; each symbol represents one mouse. LC-intact TCRβ−/−δ−/−.NLC mice have increased p53 island density and area compared to LC-deficient TCRβ−/−δ−/−.DTA mice following 9wks UVB; ***P<0.001. (c) CD207+ LC density is increased in association with p53 islands. (d, e) Epidermal cells prepared from untreated vs 5 wk chronic UVB treated TCRβ−/−δ−/−.NLC and TCRβ−/−δ−/−.DTA mice were examined for changes in gene expression (relative to untreated TCRβ−/−δ−/−.DTA) by qRT-PCR; each symbol represents one mouse,**P<0.01, ***P<0.001, Holm-Sidak correction for multiple comparisons.
Figure 5
Figure 5. Epidermal innate lymphoid cells (ILC) produce IL-22 in response to chronic UVB exposure in the presence of LC
(a) Epidermal ILC were purified from untreated and chronic UVB treated (9 wks, 400 J/m2, 3x/wk) TCRβ−/−δ−/−.NLC and TCRβ−/−δ−/−.DTA mice by flow cytometric sorting and changes in gene expression (relative to untreated TCRβ−/−δ−/−.DTA) assessed by qRT-PCR; each symbol represents one mouse, **P<0.01, ***P<0.001, Holm-Sidak correction for multiple comparisons. (b) Epidermal cells from individual untreated and 9 wk chronic UVB treated TCRβ−/−δ−/−.NLC and TCRβ−/−δ−/−.DTA mice were stimulated with PMA + Ionomycin IL-22 production assessed by flow cytometry. Top panels show CD45+ MHCII− gating, lower panels are gated on CD45+ MHCII− cells. Plots are representative of n=3 mice/group. (c) Percentage of IL-22 producing CD45+ MHCII− epidermal cells; each symbol represents one mouse, **P<0.01.
Figure 6
Figure 6. LC contribute to the epidermal stress response in a T cell independent manner
(a) Epidermal hypertrophy is greater in LC-intact TCRβ−/−δ−/−.NLC than in LC-deficient TCRβ−/−δ−/−.DTA mice following chronic UVB exposure (400 J/m2, 3x/wk, 5wks) as evaluated by measuring the minimal epidermal thickness. (b) Representative images of (a). Transepidermal waterloss (TEWL) is equivalent at baseline, but increased in LC-intact mice 48hr after tapestripping (c) as well as 4 days after epidermal abrasion by repetitive razor shaving (d). (e) Representative images of (d). Ear thickness following a single TPA application (f) and minimal epidermal thickness following twice weekly TPA for 4 wks (g) are greater in the presence of LC. Each symbol represents one mouse in panels a, c, d and f; n=7 NLC and 7 DTA in panel g.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Aberer W, Romani N, Elbe A, et al. Effects of physicochemical agents on murine epidermal Langerhans cells and Thy-1-positive dendritic epidermal cells. J Immunol. 1986;136:1210–6. - PubMed
    1. Ahlfors H, Morrison PJ, Duarte JH, et al. IL-22 fate reporter reveals origin and control of IL-22 production in homeostasis and infection. J Immunol. 2014;193:4602–13. - PMC - PubMed
    1. Artis D, Spits H. The biology of innate lymphoid cells. Nature. 2015;517:293–301. - PubMed
    1. Bobr A, Igyarto BZ, Haley KM, et al. Autocrine/paracrine TGF-β1 inhibits Langerhans cell migration. Proc Natl Acad Sci U S A. 2012;109:10492–7. - PMC - PubMed
    1. Briso EM, Guinea-Viniegra J, Bakiri L, et al. Inflammation-mediated skin tumorigenesis induced by epidermal c-Fos. Genes Dev. 2013;27:1959–73. - PMC - PubMed

Publication types

MeSH terms