cam a22 7 4500 357778456 CHVBK 20201009193440.0 cr || |||||||| 160304s2016 sz sm 00 0 eng d oai:infoscience.epfl.ch:214601 urn:nbn:ch:bel-epfl-thesis6804-9 urn 10.5075/epfl-thesis-6804 doi (SNL)991017986931103976 (NEBIS)010572125 (Sz)991017986931103976 (Sz)001874237 Sz ger rda 660 23sdnb Julier Ziad 1985- (DE-588)1085144631 Verfasser aut Engineering the Extra Domain A of fibronectin as a vaccine adjuvant and study of its role in tissue regeneration Ziad Julier Lausanne 2016 1 Online-Ressource (125 Seiten) Thèse no 6804 EPF Lausanne, 2016 application/pdf The extracellular matrix (ECM) protein fibronectin (FN) is a remarkably multifaceted molecule, including numerous fibronectin type III (FNIII) repeats carrying out different functions, which despite being extensively studied for over thirty years still presents certain functions that remain incompletely elucidated. The alternatively spliced FN variant containing the extra domain A (FNIII EDA), located between FNIII 11 and FNIII 12, is expressed in sites of injury, chronic inflammation, and solid tumors. Although its function is not well understood, FNIII EDA is known to agonize Toll-like receptor 4 (TLR4) and appears to be involved in the progression of different skin diseases while studies also suggest that FNIII EDA is involved in tissue repair. The aim of this thesis is thus to better understand some of the immunological and regenerative functions of FNIII EDA and to translate these findings to develop new vaccination strategies based on endogenous proteins. One important task in vaccinology is to develop delivery strategies by which to present antigen, along TLR agonists, so as to induce a desirable adaptive immune response. The use of TLR agonists is associated with sometimes severe constraints and concerns about safety, causing inflammatory responses. Thus, the development of safe and effective vaccines with fewer side effects is a strong need and requires the development of new adjuvants with a correct balance between a strong activation of the immune system and a low toxicity for the patient. Here, by producing various FN fragments containing FNIII EDA, we show that the immunological activity of FNIII EDA depends on its local intramolecular context within the FN chain. N-terminal extension of the isolated FNIII EDA with its neighboring FNIII repeats enhanced its activity in agonizing TLR4, while C-terminal extension abrogated it. In addition, we reveal that an elastase 2 cleavage site is present between FNIII EDA and FNIII 12. Moreover, as FNIII EDA is naturally present within the ECM, we incorporated FNIII EDA bound in a fibrin matrix to mimic the in vivo situation and create an adjuvant-rich microenvironment. Such matrices were shown to induce functional cytotoxic CD8+ T cell responses in two murine cancer models. Furthermore, combinations of specific TLR-agonists have the ability to synergize to induce immune responses that are greater than the sum of their individual effect. Hence, we demonstrate that combination of the TLR9 agonist CpG with FNIII EDA synergizes to induce an efficient immune response while keeping the dose of the toxic CpG at a minimum. Here, the combination of both adjuvants induced a potent immune response sufficient to slow down tumor growth in a murine tumor model as well as to break immune tolerance to eradicate circulating hepatitis B virus (HBV) in a transgenic HBV mouse model. Finally in the last part of this thesis we revealed that FNIII EDA is a growth factor binding domain of FN, with a strong affinity for the platelet-derived growth factor-AA (PDGF-AA). We also demonstrated that FNIII EDA and PDGF-AA have together the ability to enhance fibroblasts proliferation and differentiation into myofibroblasts. In addition, we explored the effects of fibrin matrices functionalized with variants of FNIII EDA on wound closure. EXTRAZELLULÄRE MATRIX + INTERZELLULARSUBSTANZ (CYTOLOGIE) ger (ETHUDK)000021645 ethudk FIBRONEKTIN (GLYKOPROTEINE) ger (ETHUDK)000021820 ethudk HYBRIDPROTEINE + REKOMBINANTE PROTEINE ger (ETHUDK)000051538 ethudk IMPFSTOFFE + VAKZINE (PHARMAZIE) ger (ETHUDK)000026790 ethudk KREBSHEMMENDE ARZNEIMITTEL (PHARMAZIE) ger (ETHUDK)000026738 ethudk Hochschulschrift (DE-588)4113937-9 gnd-content Hochschulschrift gnd-content xehelv xediss 2017 660 snl ehelv sb 2017/26 660 snl-sb u EXTRAZELLULÄRE MATRIX + INTERZELLULARSUBSTANZ (CYTOLOGIE) ger 576.318,1 nebis E1 u FIBRONEKTIN (GLYKOPROTEINE) ger 577.112.021,721 nebis E1 u IMPFSTOFFE + VAKZINE (PHARMAZIE) ger 615.371 nebis E1 u KREBSHEMMENDE ARZNEIMITTEL (PHARMAZIE) ger 615.277.3 nebis E1 u HYBRIDPROTEINE + REKOMBINANTE PROTEINE ger 577.112.4,7 nebis E1 u EXTRACELLULAR MATRIX (CYTOLOGY) eng 576.318,1 nebis E1 u MATRICE EXTRACELLULAIRE + MATRICE INTERCELLULAIRE (CYTOLOGIE) fre 576.318,1 nebis E1 u FIBRONECTINE (GLYCOPROTÉINES) fre 577.112.021,721 nebis E1 u FIBRONECTIN (GLYCOPROTEINS) eng 577.112.021,721 nebis E1 u VACCINES (PHARMACY) eng 615.371 nebis E1 u VACCINS (PHARMACIE) fre 615.371 nebis E1 u CARCINOSTATICS + ANTINEOPLASTICS + ONCOSTATICS + CANCER REMEDIES (PHARMACY) eng 615.277.3 nebis E1 u ONCOSTATIQUES + CARCINOSTATIQUES + ANTINÉOPLASTIQUES (PHARMACIE) fre 615.277.3 nebis E1 u PROTÉINES HYBRIDES + PROTÉINES RECOMBINANTES fre 577.112.4,7 nebis E1 u HYBRID PROTEINS + RECOMBINANT PROTEINS eng 577.112.4,7 nebis E1 École polytechnique fédérale de Lausanne (DE-588)1003500-X Grad-verleihende Institution dgg SNL NB001 NB999 - chb99 - Record https://infoscience.epfl.ch/record/214601 Thesis https://infoscience.epfl.ch/record/214601/files/EPFL_TH6804.pdf URN https://nbn-resolving.org/urn/resolver.pl?urn=urn:nbn:ch:bel-epfl-thesis6804-9 http://www.e-helvetica.nb.admin.ch/directAccess?callnumber=bel-epfl-thesis6804-9 BK020353 XK020053 XK020000 1 Hochschulschrift = Thèse/Mémoire 2 CF Elektron. Daten Fernzugriff=Fichier online EPF nebis E2 e-library 2015 these nebis EC 134 E01-20160815 NEBIS E16 E16 RH HB 17623 NEBIS E02 E02 E02SP ZTH 6804 SNL 100 1- Julier Ziad 1985- (DE-588)1085144631 Verfasser aut SNL 710 2- École polytechnique fédérale de Lausanne (DE-588)1003500-X Grad-verleihende Institution dgg SNL 856 40 Record https://infoscience.epfl.ch/record/214601 SNL 856 40 Thesis https://infoscience.epfl.ch/record/214601/files/EPFL_TH6804.pdf SNL 856 4- URN https://nbn-resolving.org/urn/resolver.pl?urn=urn:nbn:ch:bel-epfl-thesis6804-9 SNL 856 42 http://www.e-helvetica.nb.admin.ch/directAccess?callnumber=bel-epfl-thesis6804-9 NEBIS 700 1- Julier Ziad NEBIS 700 1- Hubbell Jeffrey Alan 1959- (DE-588)1089282184 Akademischer Betreuer dgs NEBIS 856 -- https://dx.doi.org/10.5075/epfl-thesis-6804 Online via EPFL E-Library E02 SWISSBIB 357778456 ORANGE 357778456