RESUMEN
Olive pollen is one of the most important causes of respiratory allergy, with Ole e 1 being the most clinically relevant sensitizing allergen. Peptide-based vaccines represent promising therapeutic approaches, but the use of adjuvants is required to strengthen the weak immunogenicity of small peptides. We propose the use of dendrimeric scaffolds conjugated to the T cell immunodominant epitope of Ole e 1 (OE109-130) for the development of novel vaccines against olive pollen allergy. Four dendrimeric scaffolds containing an ester/ether with nine mannoses, an ester succinimidyl linker with nine N-acetyl-glucosamine units or nine ethylene glycol units conjugated to OE109-130 peptide were designed, and their cytotoxicity, internalization pattern, and immunomodulatory properties were analyzed in vitro. None of the dendrimers exhibited cytotoxicity in humanized rat basophil (RBL-2H3), human bronchial epithelial Calu-3, and human mast LAD2 cell lines. Confocal images indicated that mannosylated glycodendropeptides exhibited lower colocalization with a lysosomal marker. Moreover, mannosylated glycodendropeptides showed higher transport tendency through the epithelial barrier formed by Calu-3 cells cultured at the air-liquid interface. Finally, mannosylated glycodendropeptides promoted Treg and IL10+Treg proliferation and IL-10 secretion by peripheral blood mononuclear cells from allergic patients. Mannosylated dendrimers conjugated with OE109-130 peptide from Ole e 1 have been identified as suitable candidates for the development of novel vaccines of olive pollen allergy.
Asunto(s)
Antígenos de Plantas/química , Dendrímeros/química , Manosa/inmunología , Olea/química , Olea/inmunología , Péptidos/inmunología , Proteínas de Plantas/química , Polen/inmunología , Rinitis Alérgica Estacional/prevención & control , Vacunas de Subunidad/inmunología , Adyuvantes Inmunológicos/química , Animales , Antígenos de Plantas/inmunología , Línea Celular Tumoral , Supervivencia Celular/inmunología , Citocinas/análisis , Citocinas/metabolismo , Células Epiteliales/inmunología , Células Epiteliales/metabolismo , Epítopos de Linfocito T/química , Epítopos de Linfocito T/inmunología , Humanos , Inmunogenicidad Vacunal , Manosa/química , Péptidos/química , Proteínas de Plantas/inmunología , Ratas , Mucosa Respiratoria/inmunología , Mucosa Respiratoria/metabolismo , Rinitis Alérgica Estacional/sangre , Rinitis Alérgica Estacional/inmunología , Linfocitos T/inmunología , Linfocitos T/metabolismoRESUMEN
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
RESUMEN
SCOPE: Glycodendropeptides (GDPs) functionalized with mannose can enhance allergen interaction with dendritic cells (DCs) via C-type lectin receptors (CLRs), modulating the immune response. They can present multiple peptides and have potential applications for diagnosis and treatment of food allergy (FA). The immune response induced by GDPs with mannose and Pru p 3 peptides (mono/tetravalent) with ester (D1 ManPrup3/D4 ManPrup3) or ether linkers (D1 Man-O- Prup3/D4 Man-O- Prup3) in lipid-transfer-protein-allergic patients and tolerant controls is analyzed. METHODS AND RESULTS: The immunological response induced by GDPs is studied by assessing monocyte-derived-DC maturation, lymphocyte proliferation, cytokine production, and basophil response by flow cytometry. Dn ManPrup3 was recognized by DCs via CLRs inducing DC maturation in all subjects. However, CCR7 expression is significantly upregulated in allergic patients compared to tolerant controls. These changes correlate with lymphocyte proliferation and specific production of Th2/Th1 cytokines in allergic patients. Moreover, D1 ManPrup3 does not induce basophil activation. CONCLUSION: Dn ManPrup3 induces changes in DC maturation and lymphocyte proliferation, indicating specific recognition via CLRs. Prup3-GDPs are recognized by immune cells, inducing a specific immune response and modulating the immunological response in FA patients. The specific geometry of D1 ManPrup3 in particular makes it a potential candidate for specific immunotherapy development.
Asunto(s)
Antígenos de Plantas/inmunología , Proteínas Portadoras/inmunología , Células Dendríticas/inmunología , Hipersensibilidad a los Alimentos/etiología , Glicopéptidos/inmunología , Manosa/inmunología , Proteínas de Plantas/inmunología , Linfocitos T/inmunología , Adulto , Basófilos/fisiología , Citocinas/biosíntesis , Femenino , Hipersensibilidad a los Alimentos/inmunología , Humanos , Lectinas Tipo C/fisiología , Activación de Linfocitos , Masculino , Persona de Mediana EdadRESUMEN
An effective specific immunotherapy should contain elements to generate specific recognition (T-cell peptides) and to modulate the immunological response towards a Th1/Treg pattern by enhancing dendritic cells (DCs). We propose a novel sublingual immunotherapy for peach allergy, using systems, that combine Prup3-T-cell peptides with mannose dendrons (D1ManPrup3 and D4ManPrup3). Peach anaphylactic mice were treated 1, 2 and 5 nM concentrations. Tolerance was assessed one/five weeks after finishing treatment by determining in vivo/in vitro parameters after challenge with Prup3. Only mice receiving D1ManPrup3 at 2 nM were protected from anaphylaxis (no temperature changes, decrease in Prup3-sIgE and -sIgG1 antibody levels, and secreting cells) compared to PBS-treated mice. Moreover, an increase of Treg-cells and regulatory cytokines (IL-10+/IFN-γ+) in CD4+-T-cells and DCs were found. These changes were maintained at least five weeks after stopping treatment. D1ManPrup3 is an effective new approach of immunotherapy inducing protection from anaphylaxis which persists after finishing treatment.
Asunto(s)
Anafilaxia/tratamiento farmacológico , Hipersensibilidad a los Alimentos/tratamiento farmacológico , Prunus persica/efectos adversos , Inmunoterapia Sublingual/métodos , Anafilaxia/inmunología , Animales , Antígenos de Plantas/efectos adversos , Antígenos de Plantas/inmunología , Células Dendríticas/efectos de los fármacos , Células Dendríticas/inmunología , Modelos Animales de Enfermedad , Hipersensibilidad a los Alimentos/inmunología , Glicosilación/efectos de los fármacos , Humanos , Ratones , Nanoestructuras/administración & dosificación , Nanoestructuras/química , Prunus persica/inmunología , Linfocitos T Reguladores/efectos de los fármacos , Linfocitos T Reguladores/inmunologíaRESUMEN
SCOPE: Food-specific immunotherapy (SIT) is a promising treatment for lipid transfer protein (LTP)-syndrome. We propose a novel sublingual-SIT (SLIT) that combines a Pru p 3 T-cell peptide and an oligodeoxyribonucleotide (ODN) with CpG motifs (ODN-CpG) as adjuvants to induce a specific Th1/Treg response. METHODS AND RESULTS: LTP-peach allergic mice were treated sublingually with a combination of a CpG sequence and mono- or tetravalent systems including a Pru p 3 peptide, D1 (Prup3) or D4 (Prup3). Mice were challenged intraperitoneally with Pru p 3 one or three weeks after SLIT and tolerance was assessed. Mice treated with D1 (Prup3)+CpG were protected from anaphylaxis after Pru p 3 challenge. They showed no change in body temperature, lower levels of Pru p 3-specific IgE and IgG1 antibodies and higher levels of sIgG2a compared to the untreated group. They had fewer IgE and IgG1 secreting cells and more sIgG2a secreting cells. Moreover, a significantly lower number of Pru p 3-specific CD4+ T cells and a higher number of Treg cells were found, alongside a Th1 cytokine pattern. These changes were maintained for three weeks after stopping treatment. CONCLUSION: D1 Prup3+CpG represents a promising SIT for food allergy. It is easily synthesized and induces protection from anaphylaxis to Pru p 3 that is maintained for at least three weeks.
Asunto(s)
Antígenos de Plantas/inmunología , Hipersensibilidad a los Alimentos/inmunología , Hipersensibilidad a los Alimentos/terapia , Proteínas de Plantas/inmunología , Prunus persica/inmunología , Inmunoterapia Sublingual , Administración Sublingual , Animales , Antígenos de Plantas/administración & dosificación , Proteínas Portadoras , Proliferación Celular , Citocinas/metabolismo , Modelos Animales de Enfermedad , Femenino , Inmunoglobulina E/inmunología , Inmunoglobulina G/inmunología , Ratones , Ratones Endogámicos BALB C , Proteínas de Plantas/administración & dosificación , Bazo/citología , Bazo/metabolismoRESUMEN
To improve the blood-brain barrier permeability of the trypanocidal lead compound 4,4'-bis(imidazolinylamino)diphenylamine (1), five N-alkoxy analogues were synthesized from bis(4-isothiocyanatophenyl)amine and N-alkoxy-N-(2-aminoethyl)-2-nitrobenzenesulfonamides following successive chemical reactions in just one reactor ("one-pot procedure"). This involved: (a) formation of a thiourea intermediate, (b) removal of the amine protecting groups, and (c) intramolecular cyclization. The blood-brain barrier permeability of the compounds determined in vitro by transport assays through the hCMEC/D3 human cell line, a well-known and characterized human cellular blood-brain barrier model, showed that the N-hydroxy analogue 16 had enhanced blood-brain barrier permeability compared with the unsubstituted lead compound. Moreover, this compound displayed low micromolar IC(50) against Trypanosoma brucei rhodesiense and Plasmodium falciparum and moderate activity by intraperitoneal administration in the STIB900 murine model of acute sleeping sickness.