Therapeutic potential of the classical pathway complement inhibitor analogue PRP-B0 in an acute inflammatory bowel disease model.

Anžič, Andreja (avtor), Oven, Irena (mentor), Aran, M. Josep (komentor)

Inflammatory bowel disease is a chronic disorder of the gastrointestinal tract that comprises two major conditions: Crohn’s disease and ulcerative colitis. The main focus of treating IBD so far was focused towards strategies which regulate the immune response. C4b-binding protein is plasma glycoprotein and a major soluble inhibitor of the classical and lectin pathways of complement activation. Our study describes a new mechanism of action in which an analogue of C4b-binding protein called PRP-B0 might affect activation of dendritic cells. We predicted that the dendritic cells treated with PRP-B0 would lower Th1 proinflammatory cytokines and increase IL-10, cytokine that inhibits Th1-immune response and, therefore, would improve intestinal damage. An experiment using C57BL/6 mice that lasted 9 days was made, where we induced a condition similar to ulcerative colitis using 2% dextran sodium sulfate at day 0. After that, animals were treated with either PRP-B0 or received a daily treatment with minocycline (reference treatment). We also had a control group, which was injected with Dulbecco's phosphate-buffered saline. After 9 days animals were sacrificed, and a disease activity index score was applied for each animal in order to evaluate the colon inflammatory status. The presence of multiple cytokines and chemokines in mouse serum samples were assessed using the Proteome profile array. Moreover, the presence of endotoxin in mice serum was evaluated. Afterwards an accurate quantification of chemokine CXCL13 in mouse serum was made through enzyme-linked immunosorbent assay. According to the results obtained, PRP-B0 could be a promising biological for inflammatory bowel disease treatment.

• Jezik: Angleški jezik
• Ključne besede: acute inflammatory bowel disease, Crohn’s disease, ulcerative colitis, PRP-B0, therapeutic potential, classical pathway complement inhibitor
• Vrsta gradiva: Magistrsko delo/naloga
• Tipologija: 2.09 - Magistrsko delo
• Organizacija: BF - Biotehniška fakulteta
• Založnik: [A. Anžič]
• Leto izida: 2020
• PID: 20.500.12556/RUL-113678
• UDK: 606:616.34:577.27(043.2)
• COBISS.SI-ID: 18307331
• Datum objave v RUL: 24.01.2020

Sekundarni jezik

• Jezik: Slovenski jezik
• Naslov: Terapevtski potencial analoga inhibitorja komplementa klasične poti PRP-B0 v modelu akutne vnetne črevesne bolezni.

Izvleček

Kronična vnetna črevesna bolezen je bolezen prebavil, ki jo delimo na dve glavni stanji: Crohnovo bolezen in ulcerozni kolitis. Do sedaj je zdravljenje črevesnih bolezni osredotočeno na strategije, ki uravnavajo imunski odziv. Vezavni protein C4b je plazemski glikoprotein in glavni topni inhibitor klasične in lektinske poti aktivacije komplementa, ki ima protivnetne in imunomodulatorne funkcije. Naša študija opisuje nov mehanizem delovanja, pri katerem bi analog vezavnega proteina C4b, imenovan PRP-B0, lahko vplival na aktivacijo dendritičnih celic. Predvidevamo, da bodo dendritične celice, zdravljene s PRP-B0, znižale Th1 proinflamatorne citokine in povečale IL-10, citokin, ki zavira Th1-imunski odziv, in s tem dosegle izboljšanje poškodovanega črevesja. Za eksperiment, ki je trajal 9 dni, smo uporabili miši C57BL/6. V tem obdobju smo z uporabo 2% natrijevega dekstran sulfata inducirali ulcerozni kolitis na dan 0. Živali smo nato zdravili bodisi s PRP-B0, bodisi dnevno z minociklinom (referenčno zdravljenje). Imeli smo tudi slepo in kontrolno skupino, kjer smo injicirali DPBS. Po 9 dneh so bile miši žrtvovane in bili narejeni rezultati indeksov aktivnosti bolezni za vsako žival, da bi ocenili stanje vnetega črevesja. V vzorcih mišjega seruma smo določili prisotnost večih citokinov in kemokinov z uporabo kompleta za določanje proteomskega profila. Prav tako smo ovrednotili prisotnost endotoksinov v serumu miši. S testom ELISA smo nato natančno določili koncentracijo CXCL13 v serumu. Sledila je ELISA za natančno določanje koncentracije CXCL13 v vzorcu črevesja testiranih miši. Glede na pridobljene rezultate bi bila PRP-B0 lahko obetavna molekula za biološko zdravljenje vnetnih črevesnih bolezni.

• Ključne besede: bolezen akutnega vnetja črevesja, Crohnova bolezen, ulcerozni kolitis, terapevtski potencial, PRP-B0, inhibitor komplementa klasične poti