Depletion Of Langerhans Cells Research Articles

Polymorphic photodermatosis as a disease of failed apoptotic clearance in immune-mediated inflammation and the description of a clinical case

Polymorphic photodermatosis is the most common, acquired skin disease characterized by an abnormal, recurrent, and delayed response to sunlight.
 In this article, modern views on the etiology of pathogenesis are described clinical picture, differential diagnosis, principles of treatment and prevention of polymorphic photodermatosis. The pathogenesis of this disease is a form of delayed-type hypersensitivity reaction to photoantigens. The abnormal immune response has been attributed to the immunosuppressive effects of sunlight due to a breakdown in allergen tolerance and a disruption in the mechanisms of apoptosis.
 The main symptoms of the disease are erythema and highly itchy elements of skin lesions, which are very diverse, most often represented by macules, papules and vesicles, or vesiculo-bullous, urticarial, hemorrhagic rashes, hence the name of the disease polymorphic photodermatosis. Usually one morphology prevails in one patient. The article presents a clinical case and the results of their own observations.
 Studies of the pathogenesis of polymorphic photodermatosis have confirmed the presence of an abnormal immune response. The exact mechanism of immunosuppression caused by ultraviolet radiation and the relative contribution of UV-B and UV-A in healthy people are still unclear, but the expression of TNF-alpha, IL-4 and IL-10 and the depletion of Langerhans cells appear to be key phenomena.
 Diagnosis of the disease based on the clinical picture. The need for differential diagnosis of various forms of polymorphic photodermatosis is due to their clinical and sometimes histological similarity with other skin formations. For more accurate verification of the diagnosis, a biopsy of the examined skin area is possible, followed by histology. The histology of polymorphic photodermatosis is nonspecific, in accordance with the polymorphic clinical picture, as well as depending on the timing of the biopsy.
 Further studies of the pathological mechanisms underlying polymorphic photodermatosis may allow us to develop a more targeted approach to conservative treatment and prevention.

Major Histocompatibility Complex II Expression on Oral Langerhans Cells Differentially Regulates Mucosal CD4 and CD8 T Cells

In murine periodontitis, the T helper (Th)17 response against Porphyromonas gingivalis in cervical lymph node is abrogated by diphtheria toxin-driven depletion of Langerhans cells (LCs). We determined the impact of major histocompatibility complex class II (MHC-II) presentation in LCs on Th17 cells in the oral mucosa of mice. Using an established human-Langerin promoter-Cre mouse model, we generated LC-specific deletion of the H2-Ab1 (MHC-II) gene. MHC-II expression was ablated in 81.2% of oral-resident LCs compared with >99% of skin-resident LCs. MHC-II (LCΔMHC-II) depletion did not reduce the number of CD4 T cells nor the frequency of Th17 cells compared with that in wild-type mice. However, the frequencies of Th1 cells decreased, and Helios+ T-regulatory cells increased. In ligature-induced periodontitis, the numbers of CD4 T cells and Th17 cells were similar in LCΔMHC-II and wild-type mice. Normal numbers of Th17 cells can therefore be sustained by as little as 18.8% of MHC-II-expressing LCs in oral mucosa. Unexpectedly, oral mucosa CD8 T cells increased >25-fold in LCΔMHC-II mice. Hence, these residual MHC-II-expressing LCs appear unable to suppress the local expansion of CD8 T cells while sufficient to sustain a homeostatic CD4 T-cell response. Reducing the expression of MHC-II on specific LC subpopulations may ultimately boost CD8-mediated intraepithelial surveillance at mucosal surfaces.

A population of dermal Langerin+ dendritic cells promote the inflammation in mouse model of atopic dermatitis.

Cutaneous dendritic cells (DCs) have been implicated in the pathogenesis of atopic dermatitis (AD). However, the specific role of different subsets of DCs has not been well defined. This study aimed to investigate the contributions of Langerhans cells (LCs), resident dermal Langerin+ DCs (r-Langerin+ dDCs), and newly infiltrated inflammatory dermal Langerin+ DCs (i-Langerin+ dDCs) in an AD mouse model induced by the topical application of MC903. The result showed that depletion of i-Langerin+ dDCs in DTR mice after multiple diphtheria toxin (DT) injection significantly reduced thymic stromal lymphopoietin (TSLP) production in lesions and skin inflammation alleviation. However, depletion of LCs or r-Langerin+ dDCs didn’t resulted in significant changes in skin inflammation of DTA or single DT injection-treated DTR mice compared with the wild-type (WT) mice. DT-treated DTR-WT chimeric mice with the depletion of bone marrow (BM)-derived i-Langerin+ dDCs resulted in markedly decreased skin inflammation than controls, while PBS-treated chimeric mice (DTR-WT) with only the depletion of r-Langerin+ dDCs showed inflammation comparable to that in WT mice. Furthermore, TSLP contributed to the upregulation of Langerin expression in BM-derived DCs and promoted the maturation of Langerin+ DCs. In summary, the present study demonstrated that the newly infiltrated inflammatory dermal Langerin+ DCs were essential for AD development and local TSLP production, and TSLP further promoted the production of BM-derived i-Langerin+ dDCs, which might maintain AD inflammation.

Association of polymorphous light eruption with NOD-2 and TLR-5 gene polymorphisms.

Polymorphous light eruption (PLE) is a common, immunologically mediated, photosensitive skin disease. After ultraviolet-B (UV-B) irradiation, patients with PLE show reduced Langerhans cell (LC) depletion in the epidermis, which results in a non-suppressive microenvironment in the skin. Interestingly, severe acute graft-versus-host disease (aGvHD) occurred in stem cell transplanted patients that showed no or incomplete depletion of LCs after UVB irradiation. Genetic variation in nucleotide-binding oligomerization domain 2 (NOD-2) and toll-like receptor 5 (TLR-5) genes also confers susceptibility to aGvHD. We hypothesized that PLE is associated with genetic variation in the NOD-2 and TLR-5 genes. We investigated single-nucleotid polymorphisms (SNPs) of NOD-2 (R702W, G908R, 3020Cins) and TLR-5 (A592S, P616L, N392STOP) in skin biopsies of patients with PLE (n = 143) and in healthy controls (n = 104) using restriction fragment length polymorphism analysis. The frequency of NOD-2 alleles with the SNP R702W was significantly higher in PLE than in controls (31.8% vs. 6.3%; P < 0.0001), and homozygous carriers of this mutation were more common in PLE (27.9% vs. 0%; P < 0.0001). For SNP 3020Cins, the allele frequency (7.3% vs. 0.7%; P = 0.0025) and the number of heterozygotes (14.7% vs. 1.3%; P = 0.0019) were higher in PLE. The frequency of alleles with the N392STOP SNP of the TLR5 gene, which is associated with a truncated, non-functional receptor, was significantly higher in PLE (21% vs. 5%; 7% vs. 1% homozygotes, 28% vs. 8% heterozygotes; P < 0.0001). The other SNPs did not differ significantly. This study yielded a high frequency of functional SNPs in the NOD-2 and TLR-5 genes in PLE. The same SNPs are associated with aGvHD and there are similarities in the reaction of LCs after UVB irradiation between aGvHD and PLE. This leads to the hypothesis that patients with PLE may be more susceptible to developing GvHD after stem cell transplantation, an assumption that needs to be investigated further.

Skin delivery of hapten and neurokinin-1 receptor antagonists by microneedle arrays targets neurogenic inflammation and prevents contact dermatitis

Abstract Contact dermatitis (CD) is a chronic inflammatory disease caused by type-1 immunity. Skin exposure to haptens stimulates the secretion of Substance-P (SP) and initiates the neurogenic inflammation that intensifies CD. Neurokinin-1 receptor (NK1R)-signaling by SP or hemokinin-1 (HK1) amplifies immune responses. Nonetheless, the role and therapeutic implications of the NK1R-SP-HK1 axis in CD remain unclear. We show that SP, HK-1 and the NK1R are required for CD. Specific deletion of the NK1R in keratinocytes decreased the rapid release of IL-1β and IL-6 at the site of contact sensitization which impaired the innate and adaptive immunity of CD whereas deletion of the receptor in dendritic cells (DC) prevented only the adaptive immune response of the disease. Therefore, we hypothesized that blockade of NK1R during sensitization would be a feasible immunosuppressive intervention to treat CD. We developed a system of microneedle arrays (MNA) that co-deliver hapten and NK1R antagonists into mouse skin. This immunosuppressive approach resulted in decreased skin migration and lymph node homing of stimulatory dermal DC transporting the hapten from the sensitization site. Conversely, the immunosuppressive MNA did not affect the migration and lymph node homing of epidermal Langerhans cells (LC), and depletion of LC resulted in loss of the NK1R antagonist beneficial effects. In addition, immunosuppressive MNA caused deletion of hapten-specific T cells and increased T-regulatory cells, which prevented CD-onset and -relapses in a hapten-specific manner. Our findings indicate that immune-regulation by engineering localized skin neuroimmune-networks can be used to treat cutaneous diseases that, like CD are caused by type-1 immunity.

Dietary table grape protects against ultraviolet photodamage in humans: 1. clinical evaluation

To the Editor: In animal models, grape polyphenols protect against acute and chronic ultraviolet (UV) radiation–induced inflammation and DNA damage.1,2 In addition, topical administration of grape seed proanthocyanidin extract reduced sunburn cell formation and prevented Langerhans cell depletion in humans.3

Langerhans Cells Suppress CD8+ T Cells In Situ during Mucocutaneous Acute Graft-Versus-Host Disease

Acute graft-versus-host disease (aGVHD) induced by allogenic hematopoietic stem cell transplantation is an immunological disorder in which donor lymphocytes attack recipient organs. It has been proven that recipient nonhematopoietic tissue cells, such as keratinocytes, are sufficient as immunological targets for allogenic donor T cells, whereas Langerhans cells (LCs) are potent professional hematopoietic antigen-presenting cells existing in the target epidermis and eliminated during the early phase of mucocutaneous aGVHD. Moreover, LCs have been reported to negatively regulate various types of immune responses. Here, we present data showing that initial depletion of recipient LCs exacerbates mucocutaneous lesions in a murine model of allogenic bone marrow transplantation-induced aGVHD. Furthermore, another murine model of mucocutaneous aGVHD induced in mice with keratinocytes genetically expressing chicken ovalbumin by transfer of ovalbumin-specific CD8+ OT-I cells also showed that LC-depleted recipient mice develop aggravated mucocutaneous disease owing to decreased apoptosis of skin-infiltrating OT-I cells. Moreover, coexisting LCs directly induce apoptosis and inhibit the proliferation of OT-I cells invitro partially via B7 family proteins. Collectively, our results indicate that LCs negatively regulate mucocutaneous aGVHD-like lesions in situ by inhibiting the number of infiltrating CD8+ T cells.

Dual function of Langerhans cells in skin TSLP-promoted TFH differentiation in mouse atopic dermatitis

Atopic dermatitis (AD) is among the most common chronic inflammatory skin diseases, usually occurring early in life, and often preceding other atopic diseases such as asthma. TH2 has been believed to play a crucial role in cellular and humoral response in AD, but accumulating evidence has shown that follicular helper T cell (TFH), a critical player in humoral immunity, is associated with disease severity and plays an important role in AD pathogenesis. This study aimed at investigating how TFHs are generated during the pathogenesis of AD, particularly what is the role of keratinocyte-derived cytokine TSLP and Langerhans cells (LCs). Two experimental AD mouse models were employed: (1) triggered by the overproduction of TSLP through topical application of MC903, and (2) induced by epicutaneous allergen ovalbumin (OVA) sensitization. This study demonstrated that the development of TFHs and germinal center (GC) response were crucially dependent on TSLP in both the MC903 model and the OVA sensitization model. Moreover, we found that LCs promoted TFH differentiation and GC response in the MC903 model, and the depletion of Langerin+ dendritic cells (DCs) or selective depletion of LCs diminished the TFH/GC response. By contrast, in the model with OVA sensitization, LCs inhibited TFH/GC response and suppressed TH2 skin inflammation and the subsequent asthma. Transcriptomic analysis of Langerin+ and Langerin- migratory DCs revealed that Langerin+ DCs became activated in the MC903 model, whereas these cells remained inactivated in OVA sensitization model. Together, these studies revealed a dual functionality of LCs in TSLP-promoted TFH and TH2 differentiation in AD pathogenesis.

CD1A+ LANGERHANS CELLS ARE REDUCED IN ORAL SUBMUCOUS FIBROSIS AND ORAL SQUAMOUS CELL CARCINOMA

Oral submucous fibrosis (OSMF) affects mainly the Indian subcontinent and its manifestations are closely linked with the habit of chewing betel. Immune system cells such as Langerhans cells (LC) and dendritic cells (DC) are linked to the immunopathogenesis of various lesions and have been studied for their ability of recognizing precursor malignant cells and to destroy them. Objective: Verify the distribution of LC and immature DC in OSMF and oral squamous cell carcinoma (OSCC). Study design: Fourteen cases of OSFM, 9 of OSCC associated to OSMF, and 40 of OSCC were retrieved. The original diagnoses were confirmed by 2 oral pathologists, who further classified OSFM in grade I, II, and III. OSCC cases were classified as well, moderately, and poorly differentiated. Immunoreactions against CD1a were performed and the number of positive LC and DC were quantified. Results: Analysis revealed a significant decrease in the number of immature LC and DC in OSCC compared to OSCC-OSMF (P Conclusions: The results suggest that depletion of immature LC and DC would represent an important step for developing OSCC.

Langerhans Cells Orchestrate the Protective Antiviral Innate Immune Response in the Lymph Node

SUMMARYDuring disseminating viral infections, a swift innate immune response (IIR) in the draining lymph node (dLN) that restricts systemic viral spread is critical for optimal resistance to disease. However, it is unclear how this IIR is orchestrated. We show that after footpad infection of mice with ectromelia virus, dendritic cells (DCs) highly expressing major histocompatibility complex class II (MHC class IIhi DCs), including CD207+ epidermal Langerhans cells (LCs), CD103+CD207+ double-positive dermal DCs (DP-DCs), and CD103−CD207− double-negative dermal DCs (DN-DCs) migrate to the dLN from the skin carrying virus. MHC class IIhi DCs, predominantly LCs and DP-DCs, are the first cells upregulating IIR cytokines in the dLN. Preventing MHC class IIhi DC migration or depletion of LCs, but not DP-DC deficiency, suppresses the IIR in the dLN and results in high viral lethality. Therefore, LCs are the architects of an early IIR in the dLN that is critical for optimal resistance to a disseminating viral infection.

Antigen Uptake by Langerhans Cells Is Required for the Induction of Regulatory T Cells and the Acquisition of Tolerance During Epicutaneous Immunotherapy in OVA-Sensitized Mice.

The skin is a major immunologic organ that may induce protection, sensitization or tolerance. Epicutaneous immunotherapy (EPIT) has been proposed as an attractive strategy to actively treat food allergy and has been shown to induce tolerance in sensitized mice through the induction of Foxp3+ regulatory T cells (Tregs), especially CD62L+ Tregs. Among immune cells in the skin, dendritic cells are key players in antigen-specific immune activation or regulation. The role of different populations of skin DCs in tolerance induction remains to be elucidated. Using OVA-sensitized BALB/c mice, we demonstrated that the application of a patch containing OVA-A647 to the skin resulted in allergen uptake by Langerhans cells (LCs) and CD11b+ dermal cDC2 and subsequent migration into skin draining lymph nodes. These 2 populations induced Foxp3 expression in CD4+ cells in vitro. Only LCs induced LAP+ cells and CD62L+ Tregs. Using Langerin-eGFP-DTR mice, we analyzed the role of LCs in the mechanisms of tolerance induction by EPIT in vivo. Following complete depletion of LCs, a dramatic decrease in the number of OVA+ DCs and OVA+ CD11b+ dermal cDC2 was observed in skin draining lymph nodes 48 h after epicutaneous application. Likewise, 2 weeks of EPIT in non-depleted mice induced Foxp3+ Tregs, especially CD62L+, and LAP+ Tregs in skin draining lymph nodes and spleen, whereas no induction of Tregs was observed in LC-depleted mice. Following 8 weeks of treatment, EPIT-treated mice showed significant protection against anaphylaxis accompanied by a significant increase of Foxp3+ Tregs, especially CD62L+ Tregs, which was not seen in the absence of LCs. In summary, although both LCs and CD11b+ dermal cDC2s could induce regulatory T cells, the absence of LCs during EPIT impaired treatment efficacy, indicating their crucial role in skin-induced tolerance.

Human skin explants an in vitro approach for assessing UVB induced damage

Lifestyle changes involving frequent outdoor activities are contributing to higher exposure to harmful ultraviolet light (UVB). The acute effects of UVB irradiation on human skin was evaluated in this study using freshly excised human skin from elective surgery subjected to UVB doses (0–3.76 J/cm2). The assessment of UVB induced cellular and skin damages was undertaken at two time points immediately and 24 h post exposure using in vitro, and immunohistochemical staining techniques. The results indicated no significant loss of skin integrity or significant acute mitochondrial cellular damages in UVB exposed skin sections as measured by the MTS cytotoxicity assay. The other key markers of damage showed significant extracellular LDH membrane leakages and upregulation of inflammatory cytokines such as IL-1β. Skin integrity analysis was also undertaken using H&E, HLADR, and anti-cytokeratin antibodies. The results showed significant epidermal changes, basal cell activation and Langerhans cells depletion. The research proved the usefulness of freshly excised human skin explant model in measuring UVB damage. Furthermore, freshly excised human skin maintains the natural layering and therefore does not pose the same challenges faced by commercially available reconstructed skin in terms of higher costs and accurate mimicking of all the complex interactions observed in human skin.

Langerhans cells are required for the priming and induction of an optimal immune response against an acute DNA virus infection

Abstract Resistance to ectromelia (ECTV) virus infection is dependent on the induction of a strong type I interferon (T1-IFN) response, which our lab has recently determined is primarily mediated by inflammatory monocytes (iMo) that are recruited to the draining lymph node (dLN). However, the mechanism behind how iMo are recruited is unclear. Accumulation of iMo is dependent on the production of CCL2 and CCL7 by CD11c+MHC-IIhi migratory dendritic cells (skin-mDCs) that originate from the skin of the footpad (initial site of infection), specifically CD103+CD207+ double-positive dermal DCs (DP dermal DCs), CD103−CD207− double-negative DCs (DN dermal DCs) and Langerhans cells. The transport of skin-mDCs to the dLN is dependent on the intrinsic utilization of the TLR9-MyD88-IRF7 axis, highlighting a key role for DNA sensing mechanisms in the skin. Analysis of infected and uninfected skin-mDC subsets reveal differential effector roles between the groups, with DP dermal DCs and Langerhans cells exhibiting high expression of pro-inflammatory cytokines and chemokines. Ablation of skin-mDC recruitment to the dLN with pertussis toxin (Ptx) resulted in a reduction in iMo accumulation and subsequent T1-IFN/pro-inflammatory expression with significantly higher mortality in infected B6 mice (which are normally 100% resistant to ECTV-related mortality). Finally, depletion of Langerhans cells, but not DP dermal DCs, resulted in a significant reduction in iMo accumulation in the dLN and consequently high mortality rates, therefore indicating that Langerhans cells may play an integral role in the priming and breadth of the host immune response against acute viral infections.

Dissecting antigen processing and presentation routes in dermal vaccination strategies.

The skin is an attractive site for vaccination due to its accessibility and presence of immune cells surveilling this barrier. However, knowledge of antigen processing and presentation upon dermal vaccination is sparse. In this study we determined antigen processing routes that lead to CD8+ T cell activation following dermal DNA tattoo immunization, exploiting a model antigen that contains an immunoproteasome-dependent epitope. In agreement with earlier reports, we found that DNA tattoo immunization of wild type (WT) mice triggered vigorous responses to the immunoproteasome-dependent model epitope, whereas gene-deficient mice lacking the immunoproteasome subunits β5i/LMP7 and β2i/MECL1 failed to respond. Unexpectedly, dermal immunization both of irradiated bone marrow (BM) reconstituted mice in which the BM transplant was of WT origin, and of WT mice transplanted with immunoproteasome subunit-deficient BM induced a CD8+ T cell response to the immunoproteasome-dependent epitope, implying that both BM and host-derived cells contributed to processing of delivered model antigen. Depletion of radiation-resistant Langerhans cells (LC) from chimeric mice did not diminish tattoo-immunization induced CD8+ T cell responses in most mice, illustrating that LC were not responsible for antigen processing and CD8+ T cell priming in tattoo-immunized hosts. We conclude that both BM and non-BM-derived cells contribute to processing and cross-presentation of antigens delivered by dermal DNA tattoo immunization.

Depletion of Epidermal Langerhans Cells in the Skin Lesions of Pellagra Patients.

Pellagra is a nutrient deficiency disease caused by insufficient niacin levels. Recent studies have shown that numbers of epidermal Langerhans cells decreased in other diseases caused by nutritional deficiencies, including necrolytic migratory erythema and acrodermatitis enteropathica. Epidermal Langerhans cells are capable of modulating or even halting the inflammatory reaction. The aim of this study was to examine changes in the number of Langerhans cells and other dendritic cells, and maturation of epidermal Langerhans cells in the lesional and adjacent non-lesional skin in pellagra patients. Seven pellagra patients and 10 healthy individuals who served as controls were included. The number and distribution of dendritic cells and other cutaneous cells were examined by immunohistochemistry. Epidermal Langerhans cells decreased considerably in the skin lesions of pellagra patients, whereas other dendritic cells did not change. The decrease in the number of Langerhans cells was positively correlated with the histological severity of skin lesions. As the number of Langerhans cells was not reduced in the undisturbed neighboring skin, the depletion of epidermal Langerhans cells did not precede skin damage but was a cause of prolonged severe inflammation.

Critical Role for Skin-Derived Migratory DCs and Langerhans Cells in TFH and GC Responses after Intradermal Immunization

Intradermal delivery of antigen represents a potent route of immunization that involves multiple blood- and skin-derived dendritic cell subpopulations endowed with specialized functions and dynamics in their ability to prime naïve CD4+ T cells in the draining lymph nodes. However, their individual contributions to the generation of CD4+ T follicular helper (TFH) cells and germinal centers (GCs) remain to be understood. We found that intradermal immunization of mice with a particle-based vaccine induced robust TFH and germinal center B-cell responses in skin draining lymph nodes, which were completely abrogated when skin cell emigration was prevented. However, in this later condition, both lymph node-resident and blood-derived inflammatory cells access the antigen in the draining lymph nodes but are not able to induce TFH cell differentiation. Rather, only skin-derived dendritic cells up-regulated key genes related to TFH cell development in the draining lymph nodes. Depletion of Langerhans cells partially abrogated TFH and germinal center B-cell responses. Thus, after intradermal immunization, only skin-derived migratory dendritic cells, including Langerhans cells, permit the generation of TFH cells and germinal centers. Identifying the relative contributions of tissue and lymphoid organ dendritic cell subsets in generating humoral immune responses is of great importance for the development of tailored vaccines.

Langerhans cells and NK cells cooperate in the inhibition of chemical skin carcinogenesis

ABSTRACTTissue immunosurveillance is an important mechanism to prevent cancer. Skin treatment with the carcinogen 7,12-dimethylbenz(a)anthracene (DMBA), followed by the tumor promoter 12-O-tetra-decanoyl-phorbol-13-acetate (TPA), is an established murine model for squamous cell carcinoma (SCC). However, the innate immunological events occurring during the initiation of chemical carcinogenesis with DMBA remain elusive. Here, we discovered that natural killer (NK) cells and Langerhans cells (LC) cooperate to impair this oncogenic process in murine skin. The depletion of NK cells or LC caused an accumulation of DNA-damaged, natural killer group 2D-ligand (NKG2D-L) expressing keratinocytes and accelerated tumor growth. Notably, the secretion of TNFα mainly by LC promoted the recruitment of NK cells into the epidermis. Indeed, the TNFα-induced chemokines CCL2 and CXCL10 directed NK cells to DMBA-treated epidermis. Our findings reveal a novel mechanism how innate immune cells cooperate in the inhibition of cutaneous chemical carcinogenesis.

Suppression of the CD8 T cell response by human papillomavirus type 16 E7 occurs in Langerhans cell-depleted mice.

Human papillomavirus (HPV) is an epitheliotropic virus that is the primary causal agent for cervical cancer. Langerhans cells (LC) are skin antigen presenting cells that are reduced in number in HPV-infected skin. The aim of this study was to understand the immune-modulatory effects of HPV16 E7 on LC and on the CD8 T cell response to a skin-expressed antigen. To test this, HPV16 E7 was expressed in mouse skin keratinocytes with the model antigen ovalbumin (Ova). Similar to what is observed in HPV-infected human skin, LC numbers were significantly reduced in E7-expressing mouse skin. This shows that expression of the E7 protein alone is sufficient to mediate LC depletion. Expression of E7 with Ova in keratinocytes strongly suppressed the Ova-specific CD8+ T cell response in the skin draining lymph node. When tested in LC-ablated mice, the CD8 T cell response to skin-expressed Ova in control mice was not affected, nor was the T cell response to Ova restored in E7-expressing skin. These data indicate a role for E7 in regulation of LC homeostasis in the skin and in suppression of antigen specific CD8 T cell expansion, but suggest that these two effects occur independent of each other.

Langerhans cells and CD11b+ dermal dendritic cells are involved in skin Treg generation

Abstract Antigen applied to the skin can induce immune tolerance in naïve mice and can be used as epicutaneous immunotherapy (EPIT) to treat experimental food allergy. In previous unpublished work, we have shown that EPIT induces Tregs, including a population of intestinal-homing LAP+ Tregs that can suppress mast cell activation and anaphylaxis. In this extension of that work, our goal was to determine how dendritic cells (DCs) in the skin contribute to the induction of LAP+ Tregs. Epicutaneous antigen exposure of mice that have received DO11.10 T cells with Viaskin®-OVA patches for 48 h induced OVA-specific LAP+ Tregs to a similar extent in naïve and sensitized mice. Therefore all additional studies were done in naïve mice. FACS sorting of DC subsets from skin-draining lymph nodes of Viaskin®-treated mice and then co-cultured with DO11.10 T cells demonstrated that CD11b+ CD103− dermal DCs and CD11b+EpCAM+ Langerhans cells (LCs) were both capable of antigen capture and presentation to CD4+ T cells, as well as Treg generation. PDL2 was found on both subsets and effectively discriminated between cells with and without antigen capture and presentation capacity. PDL2+ DCs presented a more mature phenotype, with higher expression of MHCII, CD86 and CD80 than PDL2− DCs. PDL2+ LCs and PDL2+ CD11b+ DCs were sorted separately and expression of regulatory molecules (GM-CSF, IL-27, IL-10, TGF-b, ALDH2 and IDO) were assessed by RT-PCR. PDL2+ LCs uniquely expressed IL-27, while PDL2+ CD11b+ DCs were enriched for ALDH2, which was then confirmed by flow cytometry. In vivo depletion of LCs using anti-CSF1 did not influence generation of Tregs in vivo. These results indicate that induction of Tregs through the skin is not restricted to one DC subset.

CD1a+ and CD83+ Langerhans cells are reduced in lower lip squamous cell carcinoma.

Actinic cheilitis (AC) is a potentially malignant lesion diagnosed in the lip of patients chronically exposed to the sun that may give rise to a fully invasive lower lip squamous cell carcinoma (LLSCC). It is known that ultraviolet radiation causes dendritic cells (DCs) depletion in the epidermis, but the role of this cellular population in lip cancer progression remains uncertain. Therefore, this study investigated the distribution of DCs in normal, dysplastic and neoplastic tissues of the lower lip. Thirteen cases of lower lip mucocele, 42 of ACs and 21 of LLSCC were retrieved and original diagnoses confirmed by two oral pathologists, who further classified ACs as low- and high-risk lesions. Immunoreactions against CD1a and CD83 identified immature and mature DCs, respectively. Immature CD1a+ Langerhans cells (LCs) were significantly decreased in LLSCC when compared to morphologically normal (P < 0.009) and dysplastic epitheliums (P < 0.003), whereas mature CD83+ LCs were significantly decreased in LLSCC when compared to normal epithelium (P = 0.038). There was no significant difference between low- and high-risk ACs regarding CD1a+ and CD83+ LCs (P > 0.05), but ACs demonstrated a lower concentration of CD1a+ LCs than normal epithelium (P < 0.009). There was no significant difference in the distribution of CD1a+ and CD83+ interstitial dendritic cells (IDCs) in the connective tissue among the studied groups (P > 0.05). These results suggest that depletion of epithelial LCs, but not IDCs in the connective tissue, would represent an important step for lip cancer development.