Allergeninduzierte Transmitterfreisetzung in der menschlichen Plazenta als mögliche Rolle für die Fetale Programmierung von Allergien in utero

Verschiedene Studien zeigten, dass für die spätere Entwicklung von Allergien das Milieu in utero für den Feten eine entscheidende Rolle spielt, da hier der Fötus bereits Allergenen ausgesetzt ist. Eine deziduale Zytokin- und Histaminproduktion der Mastzellen kann die Entstehung von Immunantworten beeinflussen. Genaue intraplazentare Mechanismen und verantwortliche Mediatoren sind noch unbekannt. Methoden und Materialien: In dieser Dissertation untersuchten wir Unterschiede in der Zytokinproduktion und Transmitterfreisetzung der Plazenta nach Kontakt mit einem relevanten Allergen bei allergischen und nichtallergischen Müttern. Hierzu entwickelten wir ein Plazentaperfusionsmodell, um simultan zwei getrennte Kotyledone einer Plazenta mit allergenenthaltendem Medium (Apfel, Hausstaub oder Milcheiweiß) einerseits und allergenfreiem Medium andererseits zu perfundieren. Zur Evaluierung dieses Modells fügten wir den Mastzell-Degranulator Compound 48/80 zum System hinzu, um zu prüfen, ob eine selektive Histaminfreisetzung aus plazentarem Gewebe von gesunden Müttern möglich ist. Die Kotyledone allergischer und gesunder Mütter wurden für 5 Stunden perfundiert, wobei aller 10 Minuten Proben aus dem Perfusionsmedium entnommen wurden. In den gesammelten Proben analysierten wir die Freisetzung von IL-2, IL-4, IL-6, IL-10, TNF-α und IFN- mittels eines Cytometric Bead Array. Die Histaminkonzentration wurde nach Extraktion und Bindung an o-Phthalaldehyd per Spektofluorometrie bestimmt. Ergebnisse: Nach Zugabe von Apfelallergen und in geringerem Maße auch nach Haustaubmilbenallergen in das Perfusionsmedium konnte eine zeitabhängige Freisetzung von TNF-α, IL-6 und in geringerem Maße auch IL-10 wesentlich früher und zu höheren Konzentrationen bei allergischen Müttern gemessen werden. In der Expression von anderen Interleukinen und Histamin ergaben sich keine signifikanten Änderungen. Allergenkontakt ruft in der Plazenta eine veränderte Zytokinsekretion hervor und könnte somit Einfluss auf die Reifung des kindlichen Immunsystems nehmen oder auch zu Störungen im immunologischen Gleichgewicht zwischen Mutter und Fetus führen

Placental immune response to apple allergen in allergic mothers

The immunological milieu in the placenta may be crucial for priming the developing foetal immune system. Early imbalances may promote the establishment of immune-mediated diseases in later life, including allergies. The initial exposure to allergens seems to occur in utero, but little is known about allergen-induced placental cytokine and chemokine release. The release of several cytokines and chemokines from placenta tissue after exposure to mast cell degranulator compound 48/80 or apple allergen in placentas from allergic and healthy mothers was to be analysed. Four placentas from women with apple allergy and three controls were applied in a placental perfusion model with two separate cotyledons simultaneously perfused with and without apple allergen (Mal d 1). Two control placentas were perfused with compound 48/80. In outflow, histamine was quantified spectrophotofluorometrically, IL-2, IL-4, IL-6, IL-10, TNF and IFN-gamma by a cytometric multiplex bead array and IL-13 and CXCL10, CXCL11, CCL17 and CCL22 with an in-house multiplex Luminex assay. Compound 48/80 induced a rapid release of histamine, CXCL10, CXCL11, CCL17 and CCL22, but not of the other factors. Apple allergen induced a time-dependent release of IL-6 and TNF, but not of histamine, in placentas of women with apple allergy compared with the unstimulated cotyledon. CCL17 levels were slightly increased after allergen stimulation in control placentas. Allergens can induce placental cytokines and chemokines distinctly in allergic and healthy mothers. These mediators may affect the prenatal development of the immune system and modify the risk of diseases related to immune disorders in childhood such as allergies.Funding Agencies|Institut Danone (Haar, Germany); European Network of Excellence within the 6th Framework Programme of the European Union [512040]; Swedish Research Council [K2011-56X-21854-01-06]; Cancer and Allergy Association; Olle Engkvist Foundation</p

Placental immune response to apple allergen in allergic mothers

The immunological milieu in the placenta may be crucial for priming the developing foetal immune system. Early imbalances may promote the establishment of immune-mediated diseases in later life, including allergies. The initial exposure to allergens seems to occur in utero, but little is known about allergen-induced placental cytokine and chemokine release. The release of several cytokines and chemokines from placenta tissue after exposure to mast cell degranulator compound 48/80 or apple allergen in placentas from allergic and healthy mothers was to be analysed. Four placentas from women with apple allergy and three controls were applied in a placental perfusion model with two separate cotyledons simultaneously perfused with and without apple allergen (Mal d 1). Two control placentas were perfused with compound 48/80. In outflow, histamine was quantified spectrophotofluorometrically, IL-2, IL-4, IL-6, IL-10, TNF and IFN-gamma by a cytometric multiplex bead array and IL-13 and CXCL10, CXCL11, CCL17 and CCL22 with an in-house multiplex Luminex assay. Compound 48/80 induced a rapid release of histamine, CXCL10, CXCL11, CCL17 and CCL22, but not of the other factors. Apple allergen induced a time-dependent release of IL-6 and TNF, but not of histamine, in placentas of women with apple allergy compared with the unstimulated cotyledon. CCL17 levels were slightly increased after allergen stimulation in control placentas. Allergens can induce placental cytokines and chemokines distinctly in allergic and healthy mothers. These mediators may affect the prenatal development of the immune system and modify the risk of diseases related to immune disorders in childhood such as allergies.Funding Agencies|Institut Danone (Haar, Germany); European Network of Excellence within the 6th Framework Programme of the European Union [512040]; Swedish Research Council [K2011-56X-21854-01-06]; Cancer and Allergy Association; Olle Engkvist Foundation</p