RESUMEN
Immunotherapy has been a milestone in combatting cancer, by complementing or even replacing classic treatments like surgery, chemotherapy, radiation, and anti-hormonal therapy. In 15%-30% of breast cancers, overexpression of the human epidermal growth factor receptor 2 (Her-2/neu) is associated with more aggressive tumor development. Passive immunization/immunotherapy with the recombinantly produced Her-2/neu-targeting monoclonal antibodies (mAbs) pertuzumab and trastuzumab has been shown to effectively treat breast cancer and lead to a significantly better prognosis. However, allergic and hypersensitivity reactions, cardiotoxicity, development of resistance, lack of immunological memory which results in continuous application over a long period, and cost-intensiveness are among the drawbacks associated with this treatment. Furthermore, intrinsic or acquired resistance is associated with the application of therapeutic mAbs, leading to the disease recurrence. Conversely, these drawbacks could be potentially overcome by vaccination, i.e. an active immunization/immunotherapy approach by activating the patient's own immune system to target cancer, along with inducing immunological memory. This review aims to summarize the main approaches investigated and undertaken for the production of Her-2/neu vaccine candidates, with the main focus on peptide-based vaccines and their evaluation in clinical settings.
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Vacunas contra el Cáncer , Vacunas contra el Cáncer/uso terapéutico , Humanos , Recurrencia Local de Neoplasia/tratamiento farmacológico , Péptidos , Vacunación , Vacunas de Subunidad/farmacología , Vacunas de Subunidad/uso terapéuticoRESUMEN
Among the mechanisms by which tumor cells escape the immune surveillance, one is the interaction between programmed cell death protein 1 (PD-1) and its ligand programmed death-ligand 1 (PD-L1). Inhibition of the PD-1/PD-L1 pathway with monoclonal antibodies as immune checkpoint inhibitors targeting PD-1 or its ligand, PD-L1, represents a milestone in cancer therapy. The application of these antibodies, however, suffers from drawbacks including failure to show a response or benefit in a majority of patients following monotherapy or combination therapy, their frequent administration, and cost intensiveness. Small peptides capable of interfering with PD-1/PD-L1 interaction represent interesting alternatives to antibody-based immune checkpoint inhibitors. Moreover, peptides representing PD-1 or PD-L1 sequences can be used in active immunization approaches to induce antibodies that enhance antitumor immunity by effectively preventing PD-1-mediated inhibition in the host. Importantly, such peptides can readily be combined with peptides derived from cancer antigens to effectively induce an antitumor immune response. In this review, we have summarized the recent developments in the use of small molecules and peptides either to directly block PD-1/PD-L1 interaction, or in vaccination approaches to induce antibody responses stimulating anticancer immunity by blocking PD-1-mediated T-cell inhibition.
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Antineoplásicos Inmunológicos , Vacunas contra el Cáncer , Neoplasias , Receptor de Muerte Celular Programada 1 , Anticuerpos Monoclonales/farmacología , Antineoplásicos Inmunológicos/farmacología , Humanos , Neoplasias/tratamiento farmacológico , Receptor de Muerte Celular Programada 1/antagonistas & inhibidores , VacunaciónRESUMEN
INTRODUCTION: Inaccurate information leads to increased scepticism concerning vaccinations among health care workers. Therefore, a proper education of medical students on vaccination is important. METHODS: During summer term 2015, we performed a paper-based survey to identify the knowledge and attitudes of medical students on vaccinations against measles, influenza and HPV in seven medical schools in Germany, Austria and Switzerland. RESULTS: Altogether, 3,652 questionnaires were analyzed. Knowledge of country-specific public recommendations increased significantly with the number of semesters of medical studies. Concerning the knowledge about vaccinations against measles, influenza and HPV, one third of the answers were given correctly. Again, a strong correlation between the knowledge and the semester of medical studies could be observed. The attitudes concerning vaccinations in general and especially for HCWs were highly positive. CONCLUSIONS: This study provides some important arguments for the development of a comprehensive vaccination education for medical students.
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Evaluación Educacional , Conocimientos, Actitudes y Práctica en Salud , Promoción de la Salud/organización & administración , Gripe Humana/prevención & control , Sarampión/prevención & control , Infecciones por Papillomavirus/prevención & control , Estudiantes de Medicina , Vacunación , Adulto , Femenino , Personal de Salud , Humanos , Masculino , Encuestas y Cuestionarios , Adulto JovenRESUMEN
Allergic poly-sensitization affects a large number of allergic patients and poses a great challenge for their treatment. In this study we evaluated the effects of the probiotic Escherichia coli Nissle 1917 (EcN) expressing a birch and grass pollen allergen chimera 'Bet v 1, Phl p 1 and Phl p 5' (EcN-Chim) on allergy prevention after oral or intranasal application in poly-sensitized mice. In contrast to oral application, intranasal pretreatment with EcN-Chim prior to poly-sensitization led to a significant reduction of lung inflammation (eosinophils, IL-5, and IL-13 in bronchoalveolar lavage) along with suppressed levels of allergen-specific serum IgE. The suppression was associated with increased levels of allergen-specific IgA in lungs and serum IgG2a along with increased Foxp3, TGF-ß, and IL-10 mRNA in bronchial lymph nodes. In vitro EcN induced high levels of IL-10 and IL-6 in both lung and intestinal epithelial cells. Importantly, using in vivo imaging techniques we demonstrated that intranasally applied EcN do not permanently colonize nose, lung, and gut and this strain might therefore be a safe delivery vector against allergy in humans. In conclusion, our data show that intranasal application of recombinant EcN expressing a multiallergen chimera presents a novel and promising treatment strategy for prevention of allergic poly-sensitization.
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Escherichia coli/genética , Vectores Genéticos/genética , Hipersensibilidad/inmunología , Membrana Mucosa/fisiología , Alérgenos/genética , Alérgenos/inmunología , Animales , Antígenos de Plantas/genética , Antígenos de Plantas/inmunología , Betula/inmunología , Reacciones Cruzadas , Citocinas/genética , Citocinas/metabolismo , Femenino , Humanos , Inmunización , Inmunoglobulina E/sangre , Ratones , Ratones Endogámicos BALB C , Microorganismos Modificados Genéticamente , Proteínas de Plantas/genética , Proteínas de Plantas/inmunología , Poaceae/inmunología , Polen/inmunología , Proteínas Recombinantes de Fusión/genética , Proteínas Recombinantes de Fusión/inmunología , VacunaciónRESUMEN
Following the publication of this article, the authors have requested that the Acknowledgements section be amended to include the financing source of the study. The correct Acknowledgments should be as follows: We gratefully acknowledge funding from the Austrian Science Fund SFB F46 and DK MCCA W1248-B30, OeAD-GmbH grants (FR13/2016) and from the Amadeus partnership Hubert Curein French and Austrian program. We thank the BioImaging Center of Lille (Frank Lafont) for the use of the IVIS Lumina XR. We gratefully thank Katharina Ambroz, Elke Korb, Karin Baier, Erika Garner-Spitzer, Joshua Tobias, Gwenaëlle Verbrugghe, and Jéremy Desramaut for their technical assistance and helpful discussions.
RESUMEN
BACKGROUND: Trees belonging to the order of Fagales show a distinct geographical distribution. While alder and birch are endemic in the temperate zones of the Northern Hemisphere, hazel, hornbeam and oak prefer a warmer climate. However, specific immunotherapy of Fagales pollen-allergic patients is mainly performed using birch pollen extracts, thus limiting the success of this intervention in birch-free areas. OBJECTIVES: T cells are considered key players in the modification of an allergic immune response during specific immunotherapy (SIT), therefore we thought to combine linear T cell epitope-containing stretches of the five most important Fagales allergens from birch, hazel, alder, oak and hornbeam resulting in a Fagales pollen hybrid (FPH) molecule applicable for SIT. METHODS: A Fagales pollen hybrid was generated by PCR-based recombination of low IgE-binding allergen epitopes. Moreover, a structural-variant FPH4 was calculated by in silico mutagenesis, rendering the protein unable to adopt the Bet v 1-like fold. Both molecules were produced in Escherichia coli, characterized physico-chemically as well as immunologically, and tested in mouse models of allergic sensitization as well as allergy prophylaxis. RESULTS: Using spectroscopic analyses, both proteins were monomeric, and the secondary structure elements of FPH resemble the ones typical for Bet v 1-like proteins, whereas FPH4 showed increased amounts of unordered structure. Both molecules displayed reduced binding capacities of Bet v 1-specific IgE antibodies. However, in a mouse model, the proteins were able to induce high IgG titres cross-reactive with all parental allergens. Moreover, prophylactic treatment with the hybrid proteins prevented pollen extract-induced allergic lung inflammation in vivo. CONCLUSION: The hybrid molecules showed a more efficient uptake and processing by dendritic cells resulting in a modified T cell response. The proteins had a lower IgE-binding capacity compared with the parental allergens, thus the high safety profile and increased efficacy emphasize clinical application for the treatment of Fagales multi-sensitization.
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Alérgenos/inmunología , Inmunoterapia , Polen/inmunología , Proteínas Recombinantes de Fusión/inmunología , Rinitis Alérgica Estacional/terapia , Tracheophyta/efectos adversos , Vacunas/inmunología , Alérgenos/química , Alérgenos/genética , Secuencia de Aminoácidos , Animales , Reacciones Cruzadas/inmunología , Células Dendríticas/inmunología , Epítopos/química , Epítopos/genética , Epítopos/inmunología , Epítopos/metabolismo , Femenino , Humanos , Inmunización , Esquemas de Inmunización , Inmunoglobulina E/inmunología , Inmunoglobulina E/metabolismo , Inmunoglobulina G/inmunología , Ratones , Datos de Secuencia Molecular , Unión Proteica/inmunología , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/genética , Rinitis Alérgica Estacional/inmunología , Alineación de Secuencia , Bazo/citología , Bazo/inmunologíaRESUMEN
BACKGROUND: The use of recombinant lactic acid bacteria (LAB) as vehicles for mucosal delivery of recombinant allergens is an attractive concept for antigen-defined allergy prevention/treatment. Interventions with LAB are of increasing interest early in life when immune programming is initiated. Here, we investigated the effect of neonatal colonization with a recombinant LAB producing the major birch pollen allergen Bet v 1 in a murine model of type I allergy. METHODS: We constructed a recombinant Lactobacillus (L.) plantarum NCIMB8826 strain constitutively producing Bet v 1 to be used for natural mother-to-offspring mono-colonization of germ-free BALB/c mice. Allergen-specific immunomodulatory effects of the colonization on humoral and cellular immune responses were investigated prior and after sensitization to Bet v 1. RESULTS: Mono-colonization with the Bet v 1 producing L. plantarum induced a Th1-biased immune response at the cellular level, evident in IFN-γ production of splenocytes upon stimulation with Bet v 1. After sensitization with Bet v 1 these mice displayed suppressed IL-4 and IL-5 production in spleen and mesenteric lymph node cell cultures as well as decreased allergen-specific antibody responses (IgG1, IgG2a, and IgE) in sera. This suppression was associated with a significant up-regulation of the regulatory marker Foxp3 at the mRNA level in the spleen cells. CONCLUSION: Intervention at birth with a live recombinant L. plantarum producing a clinically relevant allergen reduces experimental allergy and might therefore become an effective strategy for early intervention against the onset of allergic diseases.
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Antígenos de Plantas/inmunología , Inmunización , Lactobacillus plantarum/genética , Lactobacillus plantarum/inmunología , Linfocitos T Reguladores/inmunología , Células TH1/inmunología , Alérgenos/administración & dosificación , Alérgenos/genética , Alérgenos/inmunología , Animales , Animales Recién Nacidos , Antígenos de Plantas/genética , Betula/genética , Betula/inmunología , Citocinas/biosíntesis , Citocinas/inmunología , Factores de Transcripción Forkhead/genética , Factores de Transcripción Forkhead/metabolismo , Hipersensibilidad Inmediata , Isotipos de Inmunoglobulinas/sangre , Isotipos de Inmunoglobulinas/inmunología , Ratones , Ratones Endogámicos BALB C , Polen/genética , Polen/inmunología , Bazo/citología , Bazo/inmunología , Células Th2/inmunologíaRESUMEN
In travellers often several pre-departure immunizations are indicated, thus data are needed about possible interactions between vaccines. This Phase 3 study investigated the immunogenicity and safety of IC51 (JE vaccine) and HAVRIX1440 (hepatitis A vaccine) when administered alone or concomitantly to healthy subjects. The immune response was compared between single and concomitant vaccination in terms of geometric mean titre (GMT) and seroconversion rate (SCR) on Days 28 and 56. Immunogenicity was comparable for the 2 vaccines whether given together or separately which suggests that travellers to such regions could receive the vaccinations concomitantly.
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Encefalitis Japonesa/prevención & control , Vacunas contra la Hepatitis A/inmunología , Hepatitis A/prevención & control , Vacunas contra la Encefalitis Japonesa/inmunología , Adulto , Encefalitis Japonesa/inmunología , Femenino , Hepatitis A/inmunología , Anticuerpos de Hepatitis A/sangre , Anticuerpos de Hepatitis A/inmunología , Vacunas contra la Hepatitis A/administración & dosificación , Vacunas contra la Hepatitis A/efectos adversos , Humanos , Vacunas contra la Encefalitis Japonesa/administración & dosificación , Vacunas contra la Encefalitis Japonesa/efectos adversos , Masculino , Método Simple Ciego , Vacunas de Productos Inactivados/administración & dosificación , Vacunas de Productos Inactivados/efectos adversos , Vacunas de Productos Inactivados/inmunología , Adulto JovenRESUMEN
BACKGROUND: Patients with multiple sensitizations require alternative forms of treatment, as the efficacy of conventional immunotherapy is unsatisfactory. OBJECTIVE: In the present study, we sought to compare the efficacy of a subcutaneously (s.c.) and a mucosally applied polyvalent vaccine to reduce allergic immune responses within airway and lung tissues. METHODS: Female BALB/c mice were intraperitoneally immunized with recombinant (r)Bet v 1, rPhl p 1 and rPhl p 5, followed by an aerosol challenge of birch and phleum pollen extract. For tolerance induction, either a mixture of the immunodominant peptides or a hybrid peptide of the respective antigens was s.c. injected or intranasally applied before poly-sensitization. RESULTS: Mucosal but not systemic pre-treatment with poly-peptides led to significant suppression of eosinophils and IL-5 production in bronchoalveolar lavages, as well as IL-5, IL-4, IL-13 and eotaxin levels in lung cell cultures. Lung histology showed a clear reduction of cellular infiltration and mucus production only in intranasally pre-treated mice. In accordance, also the systemic immune response, characterized by IgE-dependent basophil degranulation and IL-4 levels in vitro, was significantly reduced by mucosal antigen application, but only marginally influenced by subcutaneous pre-treatment. Both treatment routes led to up-regulated CTLA4 expression in splenocytes, whereas only after mucosal pre-treatment Foxp3 expression levels were enhanced in lung CD3(+) T cells. Furthermore, intranasal but not subcutaneous application of the peptides enhanced IL-10 levels in the lungs, indicating regulatory mechanisms operating in local tolerance induction. CONCLUSION: Mucosal application of peptides is superior to systemic application in preventing both local and systemic poly-allergic T helper2 immune responses, suggesting mucosal tolerance induction as an attractive strategy for the primary and secondary prevention of allergic multi-sensitization and lung pathology.
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Alérgenos/administración & dosificación , Desensibilización Inmunológica , Hipersensibilidad/tratamiento farmacológico , Hipersensibilidad/inmunología , Inflamación/inmunología , Pulmón/inmunología , Péptidos/administración & dosificación , Alérgenos/inmunología , Animales , Basófilos/inmunología , Basófilos/metabolismo , Degranulación de la Célula , Citocinas/biosíntesis , Citocinas/inmunología , Eosinófilos/inmunología , Eosinófilos/metabolismo , Femenino , Hipersensibilidad/patología , Hipersensibilidad/prevención & control , Inmunoglobulina G/sangre , Inflamación/metabolismo , Inflamación/prevención & control , Pulmón/metabolismo , Pulmón/patología , Ratones , Ratones Endogámicos BALB C , Mucosa Nasal/inmunología , Péptidos/inmunología , Polen/inmunología , Ratas , Sistema Respiratorio/inmunología , Subgrupos de Linfocitos T/inmunología , Subgrupos de Linfocitos T/metabolismoRESUMEN
BACKGROUND: Allergen-specific immunotherapy (ASIT) is the only treatment of allergic disease that gives long-lasting relief of symptoms. However, concerns for safety and efficiency have highlighted the need for improvement of the therapy. We have previously suggested carbohydrate-based particles (CBPs) as a novel adjuvant and allergen carrier for ASIT. Our aim of this study was to evaluate the therapeutic potential of CBPs in ASIT, employing a mouse model for cat allergy. METHODS: BALB/c mice were subcutaneously immunized with the recombinant (r) cat allergen Fel d 1 followed by intranasal challenge with cat dander extract (CDE). The sensitized mice were therapeutically treated with rFel d 1 covalently coupled to CBPs (CBP-rFel d 1). Airway hyper-reactivity (AHR), infiltration of leucocytes in bronchoalveolar lavage (BAL) fluid, allergen-specific serum immunoglobulin levels and in vitro splenocyte responses were evaluated. RESULTS: Mice treated with CBP-rFel d 1 showed reduced features of allergic inflammation. They responded with (i) significantly decreased AHR and infiltration of eosinophils in BAL fluid after CDE challenge, (ii) the serum level of rFel d 1-specific IgE was reduced and the level of IgG(2)a was more pronounced after CBP-rFel d 1 treatment, and (iii) there was also a tendency of decreased allergen-specific cellular response. CONCLUSIONS: Carbohydrate-based particles are effective tools as adjuvant and allergen carriers for use in ASIT and constitutes a promising strategy to improve allergy treatment.
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Adyuvantes Inmunológicos/administración & dosificación , Carbohidratos/administración & dosificación , Desensibilización Inmunológica/métodos , Glicoproteínas/administración & dosificación , Hipersensibilidad Inmediata/terapia , Inflamación/terapia , Animales , Hiperreactividad Bronquial/inmunología , Hiperreactividad Bronquial/terapia , Carbohidratos/inmunología , Gatos , Modelos Animales de Enfermedad , Femenino , Glicoproteínas/efectos adversos , Glicoproteínas/genética , Glicoproteínas/inmunología , Humanos , Hipersensibilidad Inmediata/etiología , Hipersensibilidad Inmediata/inmunología , Inflamación/inmunología , Ratones , Ratones Endogámicos BALB C , Tamaño de la Partícula , Ratas , Proteínas Recombinantes/administración & dosificación , Proteínas Recombinantes/inmunología , Resultado del TratamientoRESUMEN
In March 2001, the European Commission funded a 3-year project (contract no. QLK3-CT-2000-00340) under the fifth Framework Programme to develop and test prototype products based on the oral delivery of vaccine and therapeutic agents using harmless lactic acid bacteria (LAB). The project, best known under its acronym LABDEL (for LAB delivery) also included research on LAB fermentation and technological innovations aimed at enhancing the efficiency of LAB delivery systems (1). One of the key scientific objectives was to investigate the possibility to prevent or treat a type I allergic disease using mucosal administration of LAB expressing the pollen allergen Bet v 1. The aim of this paper was to describe the background of the project with reference to a limited selection of articles and recent reviews as well as the results and major conclusions arising from this part of the project.
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Alérgenos/inmunología , Desensibilización Inmunológica , Hipersensibilidad Inmediata/prevención & control , Hipersensibilidad Inmediata/terapia , Lactobacillus/inmunología , Vacunas Sintéticas/inmunología , Alérgenos/genética , Animales , Antígenos de Plantas , Betula/inmunología , Europa (Continente) , Hipersensibilidad Inmediata/inmunología , Inmunidad Mucosa , Lactobacillus/genética , Ratones , Ratones Endogámicos BALB C , Vacunas Sintéticas/administración & dosificaciónAsunto(s)
Hipersensibilidad/prevención & control , Inmunoterapia/métodos , Administración Sublingual , Alérgenos/administración & dosificación , Animales , Humanos , Hipersensibilidad/inmunología , Ratones , Mucosa Bucal , Rinitis Alérgica Estacional/inmunología , Rinitis Alérgica Estacional/prevención & controlRESUMEN
BACKGROUND: As conventional immunotherapy is less efficacious in patients with allergic multi-sensitivities compared with mono-sensitized subjects, new intervention strategies are needed. Therefore, an allergen chimer was genetically engineered for treatment of multi-sensitization with birch and grass pollen on the basis of mucosal tolerance induction. METHODS: The major birch pollen allergen Bet v 1 served as a scaffold for N- and C-terminal linkage of the immunodominant peptides of the grass pollen allergens Phl p 1 and Phl p 5 and this new construct was cloned and expressed in Escherichia coli. After purification, physicochemical and immunological characterization the chimer was used for intranasal tolerance induction prior to poly-sensitization with Bet v 1, Phl p 1 and Phl p 5. RESULTS: The immunological characterization revealed that the conformation of Bet v 1 within the chimer was comparable to that of natural as well as recombinant Bet v 1. The chimer was immunogenic in mice for T and B cell responses to the three allergens. Intranasal application of the chimer prior to poly-sensitization significantly suppressed humoral and cellular allergen-specific Th2 responses and prevented development of airway inflammation upon allergen challenge. Moreover, local allergen-specific IgA antibodies were induced by the chimer. The mechanisms of poly-tolerance induction seemed to be mediated by regulatory cytokines, since TGF-beta and IL-10 mRNA in splenocytes were upregulated and tolerance was transferable with these cells. CONCLUSION: The data indicate that such allergen chimers harboring several unrelated allergens or allergen peptides could serve as mucosal polyvalent vaccines for prevention of multi-sensitivities.
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Alérgenos/uso terapéutico , Desensibilización Inmunológica , Hipersensibilidad/prevención & control , Proteínas Recombinantes de Fusión/uso terapéutico , Administración Intranasal , Alérgenos/administración & dosificación , Animales , Betula/inmunología , Femenino , Inmunidad Mucosa , Ratones , Ratones Endogámicos BALB C , Modelos Moleculares , Poaceae/inmunología , Polen/inmunología , Proteínas Recombinantes de Fusión/administración & dosificación , Proteínas Recombinantes de Fusión/química , Proteínas Recombinantes de Fusión/metabolismo , Vacunas de Subunidad/administración & dosificación , Vacunas de Subunidad/síntesis química , Vacunas de Subunidad/uso terapéuticoRESUMEN
BACKGROUND: Probiotic lactic acid bacteria (LAB) are able to modulate the host immune system and clinical trials have demonstrated that specific strains have the capacity to reduce allergic symptoms. Therefore, we aimed to evaluate the potential of recombinant LAB producing the major birch pollen allergen Bet v 1 for mucosal vaccination against birch pollen allergy. METHODS: Recombinant Bet v 1-producing Lactobacillus plantarum and Lactococcus lactis strains were constructed. Their immunogenicity was compared with purified Bet v 1 by subcutaneous immunization of mice. Intranasal application of the live recombinant strains was performed to test their immunomodulatory potency in a mouse model of birch pollen allergy. RESULTS: Bet v 1 produced by the LAB was recognized by monoclonal anti-Bet v 1 and IgE antibodies from birch pollen-allergic patients. Systemic immunization with the recombinant strains induced significantly lower IgG1/IgG2a ratios compared with purified Bet v 1. Intranasal pretreatment led to reduced allergen-specific IgE vs enhanced IgG2a levels and reduced interleukin (IL)-5 production of splenocytes in vitro, indicating a shift towards non-allergic T-helper-1 (Th1) responses. Airway inflammation, i.e. eosinophils and IL-5 in lung lavages, was reduced using either Bet v 1-producing or control strains. Allergen-specific secretory IgA responses were enhanced in lungs and intestines after pretreatment with only the Bet v 1-producing strains. CONCLUSIONS: Mucosal vaccination with live recombinant LAB, leading to a shift towards non-allergic immune responses along with enhanced allergen-specific mucosal IgA levels offers a promising approach to prevent systemic and local allergic immune responses.
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Alérgenos/inmunología , Asma/prevención & control , Lactobacillus plantarum/inmunología , Lactococcus lactis/inmunología , Proteínas de Plantas/inmunología , Vacunas/uso terapéutico , Administración Intranasal , Alérgenos/genética , Animales , Antígenos de Plantas , Asma/inmunología , Líquido del Lavado Bronquioalveolar/inmunología , Desensibilización Inmunológica , Modelos Animales de Enfermedad , Inmunidad Mucosa , Inmunoglobulina A/sangre , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Interferón gamma/inmunología , Interleucina-5/inmunología , Intestino Delgado/inmunología , Lactobacillus plantarum/genética , Lactococcus lactis/genética , Ratones , Ratones Endogámicos BALB C , Proteínas de Plantas/genética , Polen/genética , Polen/inmunologíaRESUMEN
BACKGROUND: Mucosal tolerance induction is suggested as treatment strategy for allergic diseases. Using a murine model of birch pollen (BP) allergy we investigated the long-term efficacy and the underlying mechanisms of mucosal tolerance induction with two structurally different molecules in a prophylactic and in a therapeutic set-up. METHODS: The three-dimensional major BP allergen Bet v 1 or a nonconformational hypoallergenic fragment thereof was intranasally applied before (prophylaxis) or after sensitization (therapy). RESULTS: In the prophylactic application both the Bet v 1 allergen and the fragment prevented allergic sensitization, and this effect lasted for 1 year. In the therapeutic approach established allergic immune responses were also suppressed after treatment with either of the molecules. However, a long-lasting curative effect (6 months) was only achieved with the Bet v 1 allergen but not with the Bet v 1 fragment. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis of splenocytes revealed that tolerance induction with the Bet v 1 allergen was associated with enhanced expression of transforming growth factor (TGF)-beta, interleukin (IL)-10, and Foxp3 mRNA in CD4+ T cells, whereas treatment with the fragment led to the induction of either Foxp3 (prophylaxis) or IL-10 (therapy) alone. CONCLUSION: From these data we concluded (i) that the mechanisms underlying peripheral tolerance are linked to the conformation of the antigen, (ii) that mucosal tolerance is mediated by separate regulatory cell subsets, and (iii) that the long-term efficacy of immunosuppression is associated with the presence of Foxp3+ T cells.
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Factores de Transcripción Forkhead/inmunología , Tolerancia Inmunológica , Mucosa Nasal/inmunología , Rinitis Alérgica Estacional/prevención & control , Linfocitos T/inmunología , Administración Intranasal , Alérgenos/administración & dosificación , Alérgenos/química , Alérgenos/inmunología , Animales , Betula/inmunología , Desensibilización Inmunológica , Modelos Animales de Enfermedad , Epítopos de Linfocito T/inmunología , Femenino , Factores de Transcripción Forkhead/metabolismo , Interleucina-10/biosíntesis , Interleucina-10/inmunología , Ratones , Ratones Endogámicos BALB C , Fragmentos de Péptidos/inmunología , Polen/química , Polen/inmunología , ARN Mensajero , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Linfocitos T/metabolismo , Factor de Crecimiento Transformador beta/biosíntesis , Factor de Crecimiento Transformador beta/inmunologíaRESUMEN
BACKGROUND: Almost 100 million allergic patients are sensitized to the major birch pollen allergen, Bet v 1, a 17 kDa protein containing most of the IgE epitopes present in pollens of trees belonging to the Fagales order and plant-derived food. OBJECTIVE: Our aim was to develop an approach for the rational design of B cell epitope-derived, non-allergenic peptide allergy vaccines. METHODS: According to the three-dimensional (3-D) structure of birch pollen allergen, Bet v 1, six peptides comprising 25-32 preferably solvent-exposed amino acids were synthesized. RESULTS: Because of lack of secondary structure, the peptides showed no allergenic activity in allergic patients. In a mouse model of birch pollen allergy, peptide vaccination induced Bet v 1-specific IgG and prevented IgE-mediated allergic sensitization to Bet v 1. The protective role of peptide-induced blocking antibodies is demonstrated by inhibition of allergic patients IgE binding to the allergen and by blocking of allergen-induced basophil degranulation. CONCLUSION: Our results indicate the mechanistic importance of blocking antibodies for allergy vaccination and present a B cell epitope-based approach for the rational design of safe peptide allergy vaccines whenever the structure of the disease-eliciting allergen is known.
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Alérgenos/inmunología , Betula/inmunología , Hipersensibilidad Inmediata/prevención & control , Polen/inmunología , Vacunas/inmunología , Secuencia de Aminoácidos , Animales , Antígenos de Plantas , Linfocitos B/inmunología , Basófilos/inmunología , Desensibilización Inmunológica/métodos , Ensayo de Inmunoadsorción Enzimática/métodos , Epítopos/inmunología , Liberación de Histamina/inmunología , Humanos , Inmunoglobulina E/inmunología , Inmunoglobulina G/inmunología , Ratones , Péptidos/inmunología , Estructura Terciaria de Proteína , Conejos , Ratas , Pruebas Cutáneas/métodosRESUMEN
The pathophysiological and immunological characteristics of allergic immune responses are controlled by a variety of factors. We have studied the extent to which the route of sensitization influences allergen-specific IgE synthesis and local airway inflammation using a mouse model of allergic sensitization to the major birch pollen allergen Bet v 1. Sensitization of BALB/c mice with recombinant (r)Bet v 1 was performed using intraperitoneal (i.p.), subcutaneous (s.c.) or aerosol (a.s.) sensitization protocols. Mice were analysed for allergen-specific serum antibodies by ELISA and IgE-dependent basophil degranulation. Proliferative responses and cytokine production of splenocytes were measured upon Bet v 1 stimulation in vitro. Bronchoalveolar lavages were performed after airway challenge with aerosolized birch pollen extract for assessment of eosinophilic airway inflammation and local cytokine production in vivo. Highest allergen specific IgE levels and IgE-dependent basophil degranulation were achieved using the SC route. High IL-5 production by spleen and lung cells was associated with pronounced eosinophilia in bronchoalveolar lavages. After i.p. sensitization, despite giving the highest IgG levels, only low IgE levels, basophil degranulation and IL-5 production were seen. On the other hand, a.s. sensitization, resulting in the lowest systemic IgE and IL-5 levels, led to a comparably strong airway inflammation as the s.c. route. Our finding that the route of sensitization can result in a dissociation of local and systemic immune responses may contribute to a better understanding of the pathogenesis of allergic diseases and help to develop new treatment strategies.
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Alérgenos/inmunología , Hipersensibilidad Inmediata/inmunología , Inmunoglobulina E/biosíntesis , Aerosoles , Alérgenos/administración & dosificación , Animales , Antígenos de Plantas , Basófilos/inmunología , Líquido del Lavado Bronquioalveolar/inmunología , Células Cultivadas , Citocinas/biosíntesis , Modelos Animales de Enfermedad , Eosinófilos/inmunología , Femenino , Inmunidad/inmunología , Inmunización/métodos , Inmunoglobulina E/sangre , Inmunoglobulina G/sangre , Inyecciones Intraperitoneales , Inyecciones Subcutáneas , Ratones , Ratones Endogámicos BALB C , Ratas , Bazo/inmunología , Bazo/patologíaRESUMEN
The prevalence of type I allergy to Hevea brasiliensis latex is particularly high among individuals with frequent exposure to latex products, such as health-care workers (HCW) and patients with spina bifida (SB). Treatment of latex allergy seems problematic as preventive measures, such as allergen avoidance, are not always possible and conventional immunotherapy with standardized latex extracts is not performed routinely. Thus, the aim of the present study was to establish a mouse model of latex allergy using two major latex allergens for HCWs and SB patients, Hev b 1 and Hev b 3, for sensitization. Prophylactic measures on the basis of mucosal tolerance induction with the recombinant allergens were tested in this model. Female BALB/c mice immunized intraperitoneally with recombinant (r)Hev b 1 or rHev b 3 displayed strong immune responses in vivo and in vitro. Intranasal treatment with rHev b 1 and rHev b 3 prior to sensitization led to reduced allergen-specific IgG1/IgE levels and significantly suppressed allergen-induced basophil degranulation. Moreover, lymphocyte proliferation and cytokine production (IL-4, IL-5, IFN-gamma) in vitro were significantly suppressed after pretreatment with both allergens. Suppressive cytokines, such as interleukin (IL)-10 and transforming growth factor (TGF)-beta, remained unchanged after the intranasal pretreatment, indicating mechanism of anergy rather than active immunosuppression. Taken together, these results suggest that mucosal tolerance induction with recombinant allergens could present a promising prevention strategy against latex allergy.
Asunto(s)
Alérgenos/inmunología , Tolerancia Inmunológica , Hipersensibilidad al Látex/prevención & control , Proteínas de Plantas/inmunología , Animales , Formación de Anticuerpos/inmunología , Antígenos de Plantas , División Celular/inmunología , Modelos Animales de Enfermedad , Femenino , Inmunidad Mucosa , Látex/inmunología , Hipersensibilidad al Látex/inmunología , Activación de Linfocitos/inmunología , Ratones , Ratones Endogámicos BALB C , Proteínas Recombinantes/inmunología , Bazo/inmunologíaRESUMEN
BACKGROUND: Several studies have demonstrated that mucosal administration of soluble antigens can prevent the onset or reduce the severity of certain autoimmune diseases or allergies. Few studies exist showing the efficacy of mucosal tolerance for therapy of such diseases. OBJECTIVE: The aim of the present study was to modulate an allergic immune response by intranasal antigen administration in an already sensitized organism. METHODS: A murine model of allergic asthma to birch pollen (BP) and its major allergen Bet v 1 was utilized. Sensitized mice were intranasally treated with recombinant (r)Bet v 1 in different concentrations and at different intervals. On the day the mice were killed, blood and bronchoalveolar lavage fluids were taken and immediate type I skin tests were performed. T cell proliferation and cytokine production (interleukin (IL)-5, interferon (IFN)-gamma) were measured in spleen and lung cell cultures. RESULTS: Mucosal treatment with rBet v 1 (3 x 50 microg in 4 day intervals) led to a reduction of type I skin reactions, suppressed immunoglobulin (Ig)G1/IgE antibody levels and markedly decreased IL-5 and IFN-gamma production in vitro in spleen and lung cell cultures. Moreover, lung inflammation (i.e. eosinophilia and IL-5 levels in bronchoalveolar lavage fluids) was significantly suppressed by the treatment. CONCLUSION: Our results demonstrate that intranasal treatment with rBet v 1 reduced systemic allergic immune responses as well as airway inflammation in BP-sensitized mice. We therefore suggest that mucosal tolerance induction with recombinant allergens could be a promising concept for the therapy of allergic diseases.