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Article ; Online: Recent advances in epicutaneous immunotherapy and potential applications in food allergy.

Hervé, Pierre-Louis / Dioszeghy, Vincent / Matthews, Katie / Bee, Katharine J / Campbell, Dianne E / Sampson, Hugh A

Frontiers in allergy

2023 Volume 4, Page(s) 1290003

Abstract: Given the potent immunological properties of the skin, epicutaneous immunotherapy (EPIT) emerges as a promising treatment approach for inducing immune tolerance, particularly for food allergies. Targeting the highly immunocompetent, non-vascularized ... ...

Abstract	Given the potent immunological properties of the skin, epicutaneous immunotherapy (EPIT) emerges as a promising treatment approach for inducing immune tolerance, particularly for food allergies. Targeting the highly immunocompetent, non-vascularized epidermis allows for the application of microgram amounts of allergen while significantly reducing the risk of allergen passage into the bloodstream, thus limiting systemic allergen exposure and distribution. This makes EPIT highly suitable for the treatment of potentially life-threatening allergies such as food allergies. Multiple approaches to EPIT are currently under investigation for the treatment of food allergy, and these include the use of allergen-coated microneedles, application of allergen on the skin pretreated by tape stripping, abrasion or laser-mediated microperforation, or the application of allergen on the intact skin using an occlusive epicutaneous system. To date, the most clinically advanced approach to EPIT is the Viaskin technology platform. Viaskin is an occlusive epicutaneous system (patch) containing dried native allergen extracts, without adjuvants, which relies on frequent application for the progressive passage of small amounts of allergen to the epidermis through occlusion of the intact skin. Numerous preclinical studies of Viaskin have demonstrated that this particular approach to EPIT can induce potent and long-lasting T-regulatory cells with broad homing capabilities, which can exert their suppressive effects in multiple organs and ameliorate immune responses from different routes of allergen exposure. Clinical trials of the Viaskin patch have studied the efficacy and safety for the treatment of life-threatening allergies in younger patients, at an age when allergic diseases start to occur. Moreover, this treatment approach is designed to provide a non-invasive therapy with no restrictions on daily activities. Taken together, the preclinical and clinical data on the use of EPIT support the continued investigation of this therapeutic approach to provide improved treatment options for patients with allergic disorders in the near future.
Language	English
Publishing date	2023-10-27
Publishing country	Switzerland
Document type	Journal Article ; Review
ISSN	2673-6101
ISSN (online)	2673-6101
DOI	10.3389/falgy.2023.1290003
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Article ; Online: Epicutaneous allergen immunotherapy induces a profound and selective modulation in skin dendritic-cell subsets.

Laoubi, Léo / Lacoffrette, Morgane / Valsesia, Séverine / Lenief, Vanina / Guironnet-Paquet, Aurélie / Mosnier, Amandine / Dubois, Gwendoline / Cartier, Anna / Monti, Laurine / Marvel, Jacqueline / Espinosa, Eric / Malissen, Bernard / Henri, Sandrine / Mondoulet, Lucie / Sampson, Hugh A / Nosbaum, Audrey / Nicolas, Jean-François / Dioszeghy, Vincent / Vocanson, Marc

The Journal of allergy and clinical immunology

2022 Volume 150, Issue 5, Page(s) 1194–1208

Abstract: Background: Epicutaneous immunotherapy (EPIT) protocols have recently been developed to restore tolerance in patients with food allergy. The mechanisms by which EPIT protocols promote desensitization rely on a profound immune deviation of pathogenic T- ... ...

Abstract	Background: Epicutaneous immunotherapy (EPIT) protocols have recently been developed to restore tolerance in patients with food allergy. The mechanisms by which EPIT protocols promote desensitization rely on a profound immune deviation of pathogenic T- and B-cell responses. Objective: To date, little is known about the contribution of skin dendritic cells (skDCs) to T-cell remodeling and EPIT efficacy. Methods: We capitalized on a preclinical model of food allergy to ovalbumin (OVA) to characterize the phenotype and functions of OVA<sup>+</sup> skDCs throughout the course of EPIT. Results: Our results showed that both Langerhans cells and dermal conventional cDC1 and cDC2 subsets retained their ability to capture OVA in the skin and to migrate toward the skin-draining lymph nodes during EPIT. However, their activation/maturation status was significantly impaired, as evidenced by the gradual and selective reduction of CD86, CD40, and OVA protein expression in respective subsets. Phenotypic changes during EPIT were also characterized by a progressive diversification of single-cell gene signatures within each DC subset. Interestingly, we observed that OVA<sup>+</sup> Langerhans cells progressively lost their capacity to prime CD4<sup>+</sup> T<sub>EFF</sub> cells, but gained regulatory T-cell stimulatory properties. In contrast, cDC1 were inefficient in priming CD4<sup>+</sup> T<sub>EFF</sub> cells or in reactivating T<sub>MEM</sub> cells in vitro, whereas cDC2 retained moderate stimulatory properties, and progressively biased type 2 immunity toward type 1 and type 17 responses. Conclusions: Our results therefore emphasize that the acquisition of distinct phenotypic and functional specializations by skDCs during EPIT is at the cornerstone of the desensitization process.
MeSH term(s)	Humans ; Langerhans Cells ; Desensitization, Immunologic/methods ; Food Hypersensitivity ; Ovalbumin ; T-Lymphocytes, Regulatory ; Allergens
Chemical Substances	Ovalbumin (9006-59-1) ; Allergens
Language	English
Publishing date	2022-06-30
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	121011-7
ISSN	1097-6825 ; 1085-8725 ; 0091-6749
ISSN (online)	1097-6825 ; 1085-8725
ISSN	0091-6749
DOI	10.1016/j.jaci.2022.05.025
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Article ; Online: 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.

Dioszeghy, Vincent / Mondoulet, Lucie / Laoubi, Leo / Dhelft, Véronique / Plaquet, Camille / Bouzereau, Adeline / Dupont, Christophe / Sampson, Hugh

Frontiers in immunology

2018 Volume 9, Page(s) 1951

Abstract: 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 ... ...

Abstract	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
MeSH term(s)	Allergens/immunology ; Animals ; Antigen Presentation ; Desensitization, Immunologic ; Hypersensitivity/immunology ; Hypersensitivity/pathology ; Hypersensitivity/therapy ; Langerhans Cells/immunology ; Langerhans Cells/pathology ; Mice, Inbred BALB C ; Mice, Transgenic ; Ovalbumin/immunology ; Skin/immunology ; Skin/pathology ; T-Lymphocytes, Regulatory/immunology ; T-Lymphocytes, Regulatory/pathology
Chemical Substances	Allergens ; Ovalbumin (9006-59-1)
Language	English
Publishing date	2018-09-03
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2018.01951
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Article ; Online: Epicutaneous immunotherapy protects cashew-sensitized mice from anaphylaxis.

Pelletier, Benjamin / Perrin, Audrey / Assoun, Noémie / Plaquet, Camille / Oreal, Nathalie / Gaulme, Laetitia / Bouzereau, Adeline / Labernardière, Jean-Louis / Ligouis, Mélanie / Dioszeghy, Vincent / Wavrin, Sophie / Matthews, Katie / Porcheray, Fabrice / Sampson, Hugh A / Hervé, Pierre-Louis

Allergy

2020 Volume 76, Issue 4, Page(s) 1213–1222

Abstract: Background: The prevalence of tree nut allergy has increased worldwide, and cashew has become one of the most common food allergens. More critically, cashew allergy is frequently associated with severe anaphylaxis. Despite the high medical need, no ... ...

Abstract	Background: The prevalence of tree nut allergy has increased worldwide, and cashew has become one of the most common food allergens. More critically, cashew allergy is frequently associated with severe anaphylaxis. Despite the high medical need, no approved treatment is available and strict avoidance and preparedness for prompt treatment of allergic reactions are considered dual standard of care. In the meantime, Phase III study results suggest investigational epicutaneous immunotherapy (EPIT) may be a relevant and safe treatment for peanut allergy and may improve the quality of life for many peanut allergic children. Objective: We aimed to evaluate the capacity of EPIT to provide protection against cashew-induced anaphylaxis in a relevant mouse model. Methods: The efficacy of EPIT was evaluated by applying patches containing cashew allergens to cashew-sensitized mice. As negative control, sham mice received patches containing excipient. Following treatment, mice were challenged orally to cashew and anaphylactic symptoms, as well as plasmatic levels of mast-cell proteases (mMCP)-1/7, were quantified. Results: Of 16 weeks of EPIT significantly protects against anaphylaxis by promoting a faster recovery of challenged mice. This protection was characterized by a significant reduction of temperature drop and clinical symptoms, 60 minutes after challenge. This was associated with a decrease in mast-cell reactivity as attested by the reduction of mMCP-1/7 in plasma, suggesting that EPIT specifically decrease IgE-mediated anaphylaxis. Conclusion: We demonstrate that EPIT markedly reduced IgE-mediated allergic reactions in a mouse model of cashew allergy, which suggests that EPIT may be a relevant approach to treating cashew allergy.
MeSH term(s)	Allergens ; Anacardium ; Anaphylaxis/prevention & control ; Animals ; Arachis ; Desensitization, Immunologic ; Mice ; Quality of Life
Chemical Substances	Allergens
Language	English
Publishing date	2020-10-23
Publishing country	Denmark
Document type	Journal Article
ZDB-ID	391933-x
ISSN	1398-9995 ; 0105-4538
ISSN (online)	1398-9995
ISSN	0105-4538
DOI	10.1111/all.14605
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Article ; Online: Epicutaneous immunotherapy for food allergy as a novel pathway for oral tolerance induction.

Mondoulet, Lucie / Dioszeghy, Vincent / Thébault, Claude / Benhamou, Pierre-Henri / Dupont, Christophe

Immunotherapy

2015 Volume 7, Issue 12, Page(s) 1293–1305

Abstract: Epicutaneous immunotherapy is a developing technique, aiming at desensitizing patients with food allergy with less risks that oral ingestion or injection could generate. Several clinical trials have been performed and are currently running, in milk and ... ...

Abstract	Epicutaneous immunotherapy is a developing technique, aiming at desensitizing patients with food allergy with less risks that oral ingestion or injection could generate. Several clinical trials have been performed and are currently running, in milk and peanut allergy, assessing the safety of the technique and its efficacy. Preclinical models indicate a major role in the mechanisms of desensitization, for example, Tregs and epigenetic modifications.
MeSH term(s)	Administration, Cutaneous ; Adolescent ; Animals ; Child ; Child, Preschool ; Clinical Trials as Topic ; Desensitization, Immunologic/instrumentation ; Desensitization, Immunologic/methods ; Disease Models, Animal ; Epigenesis, Genetic ; Humans ; Infant ; Mice ; Milk Hypersensitivity/therapy ; Peanut Hypersensitivity/therapy ; Skin/immunology ; T-Lymphocytes, Regulatory/immunology ; Therapies, Investigational/instrumentation ; Therapies, Investigational/methods ; Transdermal Patch
Language	English
Publishing date	2015
Publishing country	England
Document type	Journal Article ; Review
ISSN	1750-7448
ISSN (online)	1750-7448
DOI	10.2217/imt.15.86
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Article ; Online: Differences in phenotype, homing properties and suppressive activities of regulatory T cells induced by epicutaneous, oral or sublingual immunotherapy in mice sensitized to peanut.

Dioszeghy, Vincent / Mondoulet, Lucie / Puteaux, Emilie / Dhelft, Véronique / Ligouis, Mélanie / Plaquet, Camille / Dupont, Christophe / Benhamou, Pierre-Henri

Cellular & molecular immunology

2017 Volume 14, Issue 9, Page(s) 770–782

Abstract: Allergen-specific immunotherapy has been proposed as an attractive strategy to actively treat food allergy using the following three different immunotherapy routes: oral (OIT), sublingual (SLIT) and epicutaneous (EPIT) immunotherapy. Regulatory T cells ( ... ...

Abstract	Allergen-specific immunotherapy has been proposed as an attractive strategy to actively treat food allergy using the following three different immunotherapy routes: oral (OIT), sublingual (SLIT) and epicutaneous (EPIT) immunotherapy. Regulatory T cells (Tregs) have been shown to have a pivotal role in the mechanisms of immunotherapy. The aim of this study was to compare the phenotype and function of Tregs induced in peanut-sensitized BALB/c mice using these three routes of treatment. We show that although EPIT, OIT and SLIT were all able to effectively desensitize peanut-sensitized mice, they induced different subsets of Tregs. Foxp3+ Tregs were induced by the three treatment routes but with greater numbers induced by EPIT. EPIT and OIT also increased the level of LAP+ Tregs, whereas SLIT induced IL-10+ cells. The suppressive activity of EPIT-induced Tregs did not depend on IL-10 but required CTLA-4, whereas OIT acted through both mechanisms and SLIT was strictly dependent on IL-10. Moreover, the three routes influenced the homing properties of induced Tregs differently, with a larger repertoire of chemokine receptors expressed by EPIT-induced Tregs compared with OIT- and SLIT- induced cells, resulting in different protective consequences against allergen exposure. Furthermore, whereas OIT- or SLIT-induced Tregs lost their suppressive activities after treatment was discontinued, the suppressive activities of EPIT-induced Tregs were still effective 8 weeks after the end of treatment, suggesting the induction of a more long-lasting tolerance. In summary, EPIT, OIT and SLIT mediated desensitization through the induction of different subsets of Tregs, leading to important differences in the subsequent protection against allergen exposure and the possible induction of tolerance.
Language	English
Publishing date	2017-09
Publishing country	China
Document type	Journal Article
ZDB-ID	2435097-7
ISSN	2042-0226 ; 1672-7681
ISSN (online)	2042-0226
ISSN	1672-7681
DOI	10.1038/cmi.2016.14
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Article ; Online: Specific epicutaneous immunotherapy prevents sensitization to new allergens in a murine model.

Mondoulet, Lucie / Dioszeghy, Vincent / Puteaux, Emilie / Ligouis, Mélanie / Dhelft, Véronique / Plaquet, Camille / Dupont, Christophe / Benhamou, Pierre-Henri

The Journal of allergy and clinical immunology

2015 Volume 135, Issue 6, Page(s) 1546–57.e4

Abstract: Background: Allergy to cow's milk increases the risk of sensitization to other foods in young children.: Objectives: We sought to evaluate the effect of early epicutaneous immunotherapy (EPIT) on further sensitization to peanut or house dust mite ( ... ...

Abstract	Background: Allergy to cow's milk increases the risk of sensitization to other foods in young children. Objectives: We sought to evaluate the effect of early epicutaneous immunotherapy (EPIT) on further sensitization to peanut or house dust mite (HDM) in a murine model of sensitization to cow's milk. Methods: BALB/c mice orally sensitized to milk were epicutaneously treated with a Viaskin patch (DBV Technologies) loaded with milk proteins for 8 weeks. Mice were then sensitized to peanut or HDM. After sensitization to peanut, mice were exposed to a peanut regimen known to induce eosinophilic esophageal inflammation. After sensitization to HDM, mice were challenged with aerosols to HDM, and airway hyperresponsiveness was evaluated by using plethysmography. Humoral response was also analyzed. The role of regulatory T (Treg) cells was evaluated by adoptively transferring Treg cells from milk EPIT-treated mice to naive mice before sensitization to peanut. Protection against anaphylaxis was also investigated. Methylation of the promoter region of transcription factors was analyzed by using PCR assays. Results: In milk-sensitized mice specific EPIT prevented further sensitization to peanut or HDM. EPIT significantly modified the humoral response, reduced TH2 cytokine levels, decreased eosinophilic esophageal infiltration, and suppressed airway hyperresponsiveness. The protective effect was sustained over 2 months. Moreover, the adoptive transfer of milk EPIT Treg cells completely prevented sensitization to peanut and peanut-induced anaphylaxis. Milk EPIT enhanced methylation of the GATA-3 promoter region. Conclusions: Our results showed that EPIT influences the natural history of allergy and reduces the risk of further sensitization through a Treg cell-dependent mechanism.
MeSH term(s)	Adoptive Transfer ; Allergens/administration & dosage ; Allergens/immunology ; Allergens/isolation & purification ; Animals ; Arachis/chemistry ; Arachis/immunology ; Bronchial Hyperreactivity/genetics ; Bronchial Hyperreactivity/immunology ; Bronchial Hyperreactivity/physiopathology ; Bronchial Hyperreactivity/prevention & control ; Child ; Cross Protection ; Cytokines/genetics ; Cytokines/immunology ; DNA Methylation ; Desensitization, Immunologic/methods ; Disease Models, Animal ; Eosinophils/drug effects ; Eosinophils/immunology ; Eosinophils/pathology ; Female ; GATA3 Transcription Factor/genetics ; GATA3 Transcription Factor/immunology ; Gene Expression Regulation/drug effects ; Humans ; Mice ; Mice, Inbred BALB C ; Milk Hypersensitivity/genetics ; Milk Hypersensitivity/immunology ; Milk Hypersensitivity/physiopathology ; Milk Hypersensitivity/therapy ; Milk Proteins/administration & dosage ; Milk Proteins/immunology ; Milk Proteins/isolation & purification ; Peanut Hypersensitivity/genetics ; Peanut Hypersensitivity/immunology ; Peanut Hypersensitivity/physiopathology ; Peanut Hypersensitivity/prevention & control ; Plethysmography ; Promoter Regions, Genetic ; Pyroglyphidae/chemistry ; Pyroglyphidae/immunology ; Skin/drug effects ; Skin/immunology ; Skin/pathology ; T-Lymphocytes, Regulatory/drug effects ; T-Lymphocytes, Regulatory/immunology ; T-Lymphocytes, Regulatory/pathology ; T-Lymphocytes, Regulatory/transplantation ; Th1-Th2 Balance/drug effects
Chemical Substances	Allergens ; Cytokines ; GATA3 Transcription Factor ; Gata3 protein, mouse ; Milk Proteins
Language	English
Publishing date	2015-06
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	121011-7
ISSN	1097-6825 ; 1085-8725 ; 0091-6749
ISSN (online)	1097-6825 ; 1085-8725
ISSN	0091-6749
DOI	10.1016/j.jaci.2014.11.028
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Article ; Online: Epicutaneous immunotherapy using a new epicutaneous delivery system in mice sensitized to peanuts.

Mondoulet, Lucie / Dioszeghy, Vincent / Vanoirbeek, Jeroen A J / Nemery, Benoit / Dupont, Christophe / Benhamou, Pierre-Henri

International archives of allergy and immunology

2011 Volume 154, Issue 4, Page(s) 299–309

Abstract: Background: Peanut allergy is a life-threatening condition for which new efficient and safe treatment is expected. We evaluated epicutaneous immunotherapy (EPIT) as a new alternative treatment for peanut allergy in sensitized mice.: Methods: Sixty ... ...

Abstract	Background: Peanut allergy is a life-threatening condition for which new efficient and safe treatment is expected. We evaluated epicutaneous immunotherapy (EPIT) as a new alternative treatment for peanut allergy in sensitized mice. Methods: Sixty BALB/c mice were sensitized by gavages with peanut protein extract (PPE) mixed with cholera toxin. An epicutaneous delivery system, coated with 100 μg PPE (Viaskin®, DBV Technologies, Paris, France), was applied to intact skin every week during 48 h (EPIT; n = 20). This group was compared with sensitized mice treated with subcutaneous immunotherapy (SCIT; n = 20), untreated sensitized mice (sham, n = 20), and naive mice (naive; n = 20). After the 8-week treatment, a histamine release test, airway hyperreactivity measurement by plethysmography, and a resistance-compliance measurement after the challenge were performed. Blood and bronchoalveolar lavage were sampled for serology, cytokines, and cytology. Results: Specific IgE (sIgE) increased after sensitization in the EPIT (0.26 μg/ml) and SCIT (0.21 μg/ml) groups and decreased after treatment (0.09 μg/ml, p < 0.001 and 0.06 μg/ml, p < 0.001, respectively). The IgG1/IgG2a ratio decreased in the EPIT and SCIT groups versus the sham group (3.7; p < 0.001 and 2.7; p < 0.01 and 15.1, respectively). At the higher metacholine concentration, enhanced pause values were lower in the EPIT and SCIT groups than in the sham group (7.29, 6.74, and 10.99, p < 0.01, respectively), and did not differ from that of the naive group (5.06). Resistance-compliance was reversed in the treated groups versus the sham group (p < 0.001). IL-4, IL-5, IL-13, eotaxin, and eosinophils were reduced in the BAL of the EPIT and SCIT groups versus the sham group (p < 0.001). Conclusion: In peanut-sensitized mice, based on biological and physiological responses, EPIT is as efficient as subcutaneous treatment which is the reference method in immunotherapy.
MeSH term(s)	Administration, Cutaneous ; Animals ; Desensitization, Immunologic/methods ; Female ; Mice ; Mice, Inbred BALB C ; Peanut Hypersensitivity/prevention & control ; Plant Extracts/administration & dosage
Chemical Substances	Plant Extracts
Language	English
Publishing date	2011
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	1108932-5
ISSN	1423-0097 ; 1018-2438
ISSN (online)	1423-0097
ISSN	1018-2438
DOI	10.1159/000321822
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Article ; Online: Intact skin and not stripped skin is crucial for the safety and efficacy of peanut epicutaneous immunotherapy (EPIT) in mice.

Mondoulet, Lucie / Dioszeghy, Vincent / Puteaux, Emilie / Ligouis, Mélanie / Dhelft, Véronique / Letourneur, Franck / Dupont, Christophe / Benhamou, Pierre-Henri

Clinical and translational allergy

2012 Volume 2, Issue 1, Page(s) 22

Language	English
Publishing date	2012-11-12
Publishing country	England
Document type	Journal Article
ZDB-ID	2630865-4
ISSN	2045-7022 ; 2045-7022
ISSN (online)	2045-7022
ISSN	2045-7022
DOI	10.1186/2045-7022-2-22
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Article: Epicutaneous Immunotherapy Compared with Sublingual Immunotherapy in Mice Sensitized to Pollen (Phleum pratense).

Mondoulet, Lucie / Dioszeghy, Vincent / Ligouis, Mélanie / Dhelft, Véronique / Puteaux, Emilie / Dupont, Christophe / Benhamou, Pierre-Henri

ISRN allergy

2012 Volume 2012, Page(s) 375735

Abstract: Background. The aim of this study was to compare the efficacy of epicutaneous immunotherapy (EPIT) to sublingual immunotherapy (SLIT) in a model of mice sensitized to Phleum pratense pollen. Methods. BALB/c mice were sensitized by sub-cutaneous route to ... ...

Abstract	Background. The aim of this study was to compare the efficacy of epicutaneous immunotherapy (EPIT) to sublingual immunotherapy (SLIT) in a model of mice sensitized to Phleum pratense pollen. Methods. BALB/c mice were sensitized by sub-cutaneous route to pollen protein extract mixed treated for 8 weeks, using sham, EPIT, or SLIT. Measurements involved the serological response and cytokine profile from reactivated splenocytes, plethysmography after aerosol challenge to pollen, cell, and cytokine contents in the bronchoalveolar lavages (BALs). Results. After immunotherapy, sIgE was significantly decreased in the treated groups compared to sham (P < 0.001), whereas sIgG2a increased with EPIT and SLIT (P < 0.001 and P < 0.005 versus sham). Reactivated splenocytes secreted higher levels of Th2 cytokines with sham (P < 0.01). Penh values were higher in sham than EPIT and SLIT. Eosinophil recruitment in BAL was significantly reduced only by EPIT (P < 0.01). Conclusion. In this model of mice sensitized to pollen, EPIT was at least as efficient as SLIT.
Language	English
Publishing date	2012-02-02
Publishing country	United States
Document type	Journal Article
ZDB-ID	2616877-7
ISSN	2090-553X ; 2090-5521
ISSN (online)	2090-553X
ISSN	2090-5521
DOI	10.5402/2012/375735
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