Dokument

Zusammenfassung:
Die antimikrobielle Wirkung von Bactericidal/permeability-increasing protein (BPI), einem Protein aus den azurophilen Granula neutrophiler Granulozyten (nGr), gegen gramnegative Bakterien ist seit Jahrzehnten bekannt. Dabei kann es sowohl LPS neutralisieren, wie auch Bakterien opsonisieren und damit deren Phagozytose verstärken. Im Rahmen dieser Arbeit konnte nun erstmals eine Funktion von BPI gegenüber grampositiven Bakterien, im speziellen Listeria monocytogenes (L.m.) nachgewiesen werden. Zum einen konnte gezeigt werden, dass humane nGr auf Stimulation mit Listerien mit einer BPI-Sekretion reagierten, zum anderen, dass BPI an die Bakterien binden konnte. Diese Bindung resultierte in einer direkten antimikrobiellen Wirkung gegen L.m.. Eine intragastrale Infektion von Wildtyp (WT)- Mäusen führte zu gesteigerten Expression der BPI-mRNA im Dünndarm, was einen ersten Hinweis auf die Funktion von BPI während einer Listeriose gab. BPI-defiziente (BPI-/-) Mäuse waren im Modell der murinen Listeriose unabhängig von der Applikationsroute suszeptibler gegenüber den Bakterien. Dies zeigte sich durch erhöhte Bakterienlasten in den infizierten Organen und eine verstärkte systemische Entzündung; wobei auch BPI-/- Mäuse letztlich in der Lage waren, die Bakterien zu kontrollieren. Um zu untersuchen welche Zellen für die erhöhte Suszeptibilität der BPI-/- Mäuse verantwortlich sind, wurden Analysen mit Makrophagen, DCs und nGr durchgeführt, die die erste Abwehrlinie bei einer Listeriose bilden. Weder isolierte Makrophagen noch DCs aus BPI-/- Mäusen konnten den in vivo Phänotyp der Mäuse rekapitulieren. Allerdings zeigten sich nach Stimulation von nGr aus WT und BPI-/- Mäusen mit UV-inaktivierten Listerien signifikante Unterschiede in der TNF- und ROS-Produktion der Zellen. BPI-/- nGr reagierten deutlich stärker auf die Stimulation. Die nGr beider Genotypen wiesen vergleichbare Effizienz bei der Phagozytose lebender Bakterien auf, aber unterschieden sich deutlich in ihrer Fähigkeit die Listerien nach 24 h abzutöten. In BPI-/- nGr fand sich nach 24 h eine dreifach höhere Bakterienlast. Aus den in vitro Daten kann auf das in vivo Modell die Hypothese übertragen werden, dass vermutlich nGr im Lauf einer Listeriose für die erhöhte Suszeptibilität der BPI-/- Mäuse verantwortlich sind. Da sich die BPI-/- Mäuse von der Infektion erholen, ist dabei davon auszugehen, dass auch in den BPI-/- Mäuse eine protektive adaptive Immunantwort ausgebildet werden kann.

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