Depletion Of Epidermal Langerhans Cells Research Articles

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.

Immunohistological assessment of cutaneous drug hypersensitivity in patients with HIV infection.

The pathogenesis of drug hypersensitivity in patients with HIV infection is unknown. To study further the nature of hypersensitivity, the histopathological features of morbilliform drug hypersensitivity reactions were examined in a group of HIV-infected patients. Skin sections from 23 HIV-infected subjects with morbilliform drug hypersensitivity reactions were examined by light microscopy, direct immunofluorescence and immunohistochemistry, to determine the nature of the inflammatory infiltrate and the role of immunoglobulin, complement and cytokines. The principal light microscopic findings were spongiosis, hydropic generation of the basal layer, civatte bodies, an epidermal lymphocytic infiltrate (48%), and a perivascular dermal infiltrate of lymphocytes (87%) and macrophages (52%). Two patients had findings consistent with toxic epidermal necrolysis. Immunohistochemistry demonstrated that the lymphocytic infiltrate consisted of CD8+, HLA-DR+ T lymphocytes (some of which also stained for CD38), a marked depletion of epidermal Langerhans cells (90%), and strong cytoplasmic staining of keratinocytes for IL-6 (60%), IL-1 beta (50%), tumour necrosis factor-alpha (TNF-alpha) (45%) and to a lesser degree, interferon-gamma (IFN-gamma) (35%). Immunofluorescence did not demonstrate any significant deposition of immunoglobulin or complement. The histological findings were independent of the responsible drug, the duration of either therapy or the rash, and of peripheral blood CD4+ and CD8+ cell counts. These findings suggest that activated CD8+ lymphocytes and perhaps epidermal production of cytokines are involved in the pathogenesis of cutaneous drug hypersensitivity in HIV-infected patients. The common histological features, regardless of the causative drug, suggest a common pathogenesis.

Nicotinic Acid-Induced Flushing Is Mediated by Activation of Epidermal Langerhans Cells

The antidyslipidemic drug nicotinic acid (niacin) has been used for decades. One of the major problems of the therapeutical use of nicotinic acid is a strong cutaneous vasodilation called flushing, which develops in almost every patient taking nicotinic acid. Nicotinic acid-induced flushing has been shown to be mediated by the nicotinic acid receptor GPR109A and to involve the formation of vasodilatory prostanoids. However, the cellular mechanisms underlying this short-term effect are unknown. Here, we show that epidermal Langerhans cells are essential for the cutaneous flushing response induced by nicotinic acid. Langerhans cells respond with an increase in [Ca(2+)](i) to nicotinic acid and express prostanoid synthases required for the formation of the vasodilatory prostanoids prostaglandin E(2) and prostaglandin D(2). Depletion of epidermal Langerhans cells but not of macrophages or dendritic cells abrogates nicotinic acid-induced flushing. These data unexpectedly identify epidermal Langerhans cells as essential mediators of nicotinic acid-induced flushing and may help to generate new strategies to suppress the unwanted effects of nicotinic acid. In addition, our results suggest that Langerhans cells besides their immunological roles are also involved in the local regulation of dermal blood flow.

Topical Imiquimod Treatment Prevents UV-Light Induced Loss of Contact Hypersensitivity and Immune Tolerance

Imiquimod (1-(2-methylpropyl)-1H-imidazo[4,5-c]quinolin-4-amine) is a TLR7 agonist that induces cytokine production in TLR7 bearing antigen-presenting cells (APCs), including IL-12, a cytokine that has been demonstrated to be a critical effector molecule for contact hypersensitivity (CHS). To test our hypothesis that topical applications of imiquimod may protect the skin immune system against the deleterious effects of UV light exposures, we treated animals with this agent, or its vehicle or nothing before UV exposures. Although topical imiquimod exposures before UV light did not prevent the depletion of epidermal Langerhans cells, it did prevent the loss of CHS. IL-12 was important in the protective role of imiquimod in preventing UV-induced loss of CHS, as systemic treatment of mice with an anti-IL-12 p70 monoclonal antibody blocked the protective effects of imiquimod. Additionally, only imiquimod-treated mice were resistant to hapten-specific tolerance induction after UV irradiation at the site of the initial sensitization with the hapten 2,4 dinitro-1-fluorobenzene. To model for the effects of TLR7 activation on the UV effect on antigen-APCs, XS52 cell line was used to study this interaction in an in vitro model system. This cell line expressed mRNA for TLR7, downregulated IkappaB, phosphorylated c-Jun N-terminal kinase, and secreted cytokines after exposure to imiquimod or lipopolysaccharide. Activation of the TLR7 signaling pathway on XS52 before UV-light exposures enhanced IL-12p70 secretion by this cell line. Similarly, activation of TLR7 on XS52 before UV-light exposure also prevented the UV-induced loss of IFN-gamma triggering in T cells during an allogeneic mixed lymphocyte reaction. Imiquimod-treated, UV-irradiated XS52 triggered a more vigorous IFN-gamma production than did either imiquimod-treated XS52 or UV-irradiated XS52, again suggesting a synergy between the two treatments. Lastly, enriched lymph node CD11c+ APCs from mice treated with UV irradiation, imiquimod alone or the combination of UV irradiation and imiquimod indicated the same in vivo synergy between imiquimod irradiation and UV irradiation in enhancing IL-12p70 production. These data suggest that topical imiquimod applications may play a role in preventing UV-induced impairment of the skin immune system, which is thought to be one of the critical events that allow the development of UV-induced skin cancers.

Pefloxacin and ciprofloxacin increase UVA-induced edema and immune suppression.

Quinolone antibiotics are popularly prescribed antibiotics because of their wide antibacterial spectrum and lowered bacterial resistance. Quinolone antibiotics are one of the well-known photosensitizers that induce phototoxicity. Their role in photocarcinogenesis has been suggested in some studies. Mice were treated with two quinolone antibiotics (ciprofloxacin, which is less phototoxic, and pefloxacin, which is more phototoxic) to study the effect of the antibiotics on sunburn and immune suppression by ultraviolet A (UVA) irradiation. The effects of a combined treatment with UVA and these quinolone antibiotics were measured on back skin swellings, sunburn cell formations, depletion of epidermal Langerhans cells, and local and systemic suppression of contact hypersensitivity. Mice treated with both UVA and quinolone showed significantly increased back skin swellings and decreased epidermal Langerhans cells than mice treated with UVA only. Sunburn cells were increased significantly in mice treated with pefloxacin and 50 J/cm2 of UVA. Combination of pefloxacin and UVA suppressed local contact hypersensitivity significantly, but not systemic contact hypersensitivity. Phototoxic quinolones augmented the effect of UVA by increasing sunburn and apoptosis, depleting Langerhans cells and suppressing local immune response. By affecting apoptosis and immune suppression, they may facilitate photocarcinogenesis caused by UVA.

Ultraviolet radiation-induced tolerance.

Ultraviolet (UV) radiation is one of the most important environmental factors. Besides its well-known advantages and its indispensable effects on human life, UV light, particularly the middle wavelength range (290-320 nm [UVB]), can harm human health by inducing skin cancer, premature skin aging, inflammation, and cell death (1-4). Over the last 25 years, it has become apparent that exposure to UVB radiation may also suppress immune reactions. Review of the numerous studies on the immunosuppressive properties of UV radiation led to two quite surprising observations: 1) that UV exposure can have systemic immunologic consequences 2) that UV radiation can induce antigen-specific tolerance. This paper briefly reviews the phenomenon of UV-induced tolerance.

Effects of UV on the migration and function of epidermal antigen presenting cells

Depletion of epidermal Langerhans cells (LC) and the concomitant depression of the skin immune system after excessive exposure to ultraviolet B light (UVB) has been established in the international literature for some time. Our investigations were intended to determine whether or not these phenomena occurred as a direct result of increased LC migration being triggered by the UVB exposure. To test this hypothesis, a sheep model was established in which the lymphatic vessels draining a defined region of skin were cannulated and the cells migrating towards the regional lymph node continuously collected. Cell populations in these collections were identified and enumerated by indirect immunofluorescence and flow cytometry. These experiments showed there was a significant, dose-dependent increase in the rate of LC migration from sheep skin after exposure to doses of UVB light exceeding 1 minimal erythemal dose (MED). In a series of parallel experiments, the functional characteristics of dendritic cells (DC) migrating from normal or UVB irradiated sheep were studied. To assay them, enriched preparations of DC were collected via cannulated afferent lymphatic vessels and pulsed with antigen prior to incubation with autologous peripheral blood lymphocytes. The relative efficiency of antigen presentation was determined by the ability of DC to induce T cell proliferation. Our data clearly demonstrate that there is a profound loss of normal antigen-presenting cell function after exposure to UVB light. Various experiments were undertaken to determine the mechanism(s) associated with these changes in migration kinetics and cellular function. Electron microscopic examinations of LC migrating from normal or UVB irradiated skin have demonstrated a profound loss of dendritic processes after UVB exposure. This provides a possible explanation for the changes in skin immunity after UVB exposure.

Commercial sunscreen lotions prevent ultraviolet radiation-induced depletion of epidermal Langerhans cells in Skh-1 and C3H mice.

There is much controversy regarding the ability of sunscreens to prevent ultraviolet (UV)-induced immune suppression. Epidermal Langerhans cells (LC) play a key antigen-presenting role in the afferent limb of the immune system's response to antigens introduced through the skin. It has been suggested that depletion of LC in UV-exposed skin is a critical step toward the induction of immunosuppression by UV radiation. There are a number of disparate reports with inconsistent results concerning the ability of sunscreens to prevent UV-induced depletion of LC. The purpose of this study was to systematically evaluate the ability of sunscreens to prevent UV-induced LC depletion in mice. Epidermal sheets obtained from skin biopsies taken from mice exposed to UV radiation from Kodacel-filtered FS20 sunlamps, which do not emit UV power at wavelengths < 290 nm, were immunoperoxidase stained for LC using a rat monoclonal antibody against mouse Ia (major histocompatibility complex class II antigen). Time course and dose-response curves for LC depletion were generated for Skh-1 and C3H mice. Dose-response curves for acute UV exposure induced depletion of LC in Skh-1 and C3H mice were similar, but not identical. LC density in the skin of Skh-1 mice that received chronic UV exposure (3 days/week for 8 weeks) was reduced by 62% after 2 weeks of exposure, but returned to normal levels by 6 weeks. Five commercial sunscreen lotions with labeled sun protection factors (SPF) of 4, 8, 15, 30 and 45 were tested for their capacity to block UV-induced depletion of LC. LC were depleted approximately 75% in the skin of unprotected or placebo lotion treated Skh-1 mice exposed to UV given on two consecutive days. Conversely, LC depletion was prevented in similarly UV exposed Skh-1 mice protected with a SPF 30 sunscreen. In C3H mice the levels of protection against LC depletion provided by the five sunscreens were proportional to the level of protection predicted by their labeled SPF. Comparisons of dose-response curves showed that significantly higher doses of UV were required for LC depletion and induction of skin edema than for the induction of local suppression of contact hypersensitivity. Thus, at UV doses where sunscreens provide complete protection against immunosuppression of contact hypersensitivity, prevention of LC depletion and skin edema would be expected.

Turnover and kinetics of epidermal Langerhans cells and their dendritic precursor cells in experimental contact dermatitis. A correlated ultrastructural-morphometric and immunohistochemical evaluation.

The numerical density of epidermal Langerhans cells (LCs) in contact sensitivity and toxic contact dermatitis is still a matter of controversy, mainly due to changes in the phenotypic markers of this antigen-presenting cell during the skin reactions. Since the electron microscopic detection of Birbeck granules is the most reliable marker for the identification of normal and pathologically altered LCs, we performed an ultrastructural-morphometric time-course analysis to evaluate their epidermal turnover in the earskin of BALB/c mice after painting the ears with the hapten 2,4-dinitrofluorobenzene and the irritant croton oil. The counts revealed degeneration and depletion of epidermal LCs in both allergic and toxic dermatitis. In contrast, a slightly increased number of activated epidermal LCs was found during contact sensitization. All experimental procedures resulted in an enhanced immigration of so-called indeterminate dendritic cells which also became ultrastructurally activated and often showed Birbeck granule-like formations at their cell membrane. Immunohistochemistry with the monoclonal antibody 4F7, a new marker for dendritic precursor cells of LCs, demonstrated a significant increase in these accessory cells in the epidermis. Our results indicate that contact sensitivity and toxic skin reactions are characterized by complex but distinct changes in the turnover, kinetics and cellular properties of epidermal LCs and their dendritic precursor cells.

Effects of crude oil and ultraviolet radiation on immunity within mouse skin

Previous studies indicate that crude oil leads to increased pigmentation and erythema (sunburn) in response to sunlight in exposed individuals. However, no information is currently available concerning whether crude oil exposure might enhance the immunosuppressive effects of solar ultraviolet radiation (UVR) on the skin. In order to address this question, the back skin of shaved, female C3H/HeN mice was exposed to crude oil with or without subsequent treatment with medium-wavelength (UVB) (200 J/m2) or long-wavelength (UVA) (20,000 J/m2) UVR. Immune function was assessed in treated mice by measuring their ability to mount contact hypersensitivity responses to a hapten (2,4-dinitro-1-flyorobenzene, DNGFB) applied to the site of crude oil and UVR treatment as determined by ear swelling upon subsequent challenge. Since Langerhans cells represent an important component of immunity within the skin and because suppression of contact hypersensitivity following UVR treatment is often accompanied by disappearance of Langerhans cells from the epidermis, the impact of these agents on epidermal Langerhans cell density was also analyzed. This was accomplished by enumerating IA-positive cells within the epidermis of treated skin. In these studies, crude oil alone induced inhibition of contact hypersensitivity but had no effect on epidermal Langerhans cells. In contrast, combined treatment with crude oil and UVA led to suppression of contact hypersensitivity, which was accompanied by depletion of epidermal Langerhans cells.

Depletion and Repopulation of Langerhans Cells in Nonsegmental Type Vitiligo

The role of Langerhans cells in the pathogenesis of nonsegmental type vitiligo is still unknown. In this study, biopsies were taken from 26 patients at various stages of nonsegmental type vitiligo and morphometrically observed to investigate the kinetics of Langerhans cells in patients at various stages of the disease. A marked depletion of OKT6-positive and ATPase-positive epidermal dendritic cells was noted in patients with active nonsegmental type vitiligo. A repopulation of both OKT6-positive and ATPase-positive epidermal dendritic cells was noted in patients with stable nonsegmental type vitiligo. Profound depletion of epidermal OKT6-positive and ATPase-positive dendritic cells was noted in patients with repigmenting nonsegmental type vitiligo receiving treatments involving topical use of 0.05% Fluocinonide cream or PUVA photochemotherapy. Transmission electron microscopy confirmed the absence of epidermal dendritic cells (Langerhans cells and intermediate cells) in patients with active and repigmenting nonsegmental type vitiligo. In active nonsegmental type vitiligo, two possible explanations are proposed for the depletion of OKT6-positive and ATPase-positive epidermal dendritic cells (presumptive Langerhans cells): 1) the cells are destroyed by cytotoxic factors released during the course of destruction of melanocytes in active vitiligo, and/or 2) they leave the epidermis and migrate to regional lymph nodes to present certain antigens which are released from certain destroyed epidermal cells (keratinocytes or melanocytes) during the course of active vitiligo. The repopulated epidermal Langerhans cells may result from phenotypically transformed dermal dendritic cells in the depigmented lesions of patients with stable vitiligo. Since various therapies which result in repigmentation deplete the density of epidermal Langerhans cells markedly, it is suggested that depletion of epidermal Langerhans cells in stable vitiligo may aid in repigmentation. It is also proposed that the repopulated epidermal Langerhans cells may play a role in inhibiting the proliferation of epidermal melanocytes in depigmented lesions of stable vitiligo, thus various methods of treatment which deplete the Langerhans cells may eventually aid in the repigmentation of nonsegmental type vitiligo.

Depletion of epidermal langerhans cells and Ia immunogenicity from tape-stripped mouse skin.

To explore the relationships among Ia antigen expression, epidermal Langerhans cells, and the immunogenicity of skin allografts, cellophane tape-stripping was used in H-2 congenic and recombinant mice of defined immunogenetic disparity. Tape-stripping of murine abdominal wall skin achieved almost complete depletion of epidermal Langerhans cells within a few hours of application, as measured by cell surface ATPase and expression of Ia antigens. Tape-stripping also reduced, to a considerable degree (but not absolutely), the Ia immunogenicity of skin allografts prepared from stripped surfaces. No comparable reduction in immunogenicity of class I major histocompatibility determinants was observed, suggesting that Langerhans cells are relatively unimportant in the presentation of H-2K antigens in skin grafts. Langerhans cells reappear within 24 h of tape-stripping to anatomically intact skin, but are detectable in orthotopically grafted only after the graft has been in residence for 4 d, i.e., shortly after it has acquired a blood supply. Repopulating Langerhans cells at that time and thereafter are exclusively of host origin. These results indicate that the traffic of Langerhans cells to the skin can be extremely dynamic, especially when the epidermal surface has been markedly disturbed, and the data imply that, under normal circumstances, large numbers of Langerhans cells can be mobilized readily from an available pool of precursors.