Tesis doctoral de María nna Di Scala
Worldwide 350 million people suffer from hbv infection and approximately 1 million people die annually because of hbv-induced liver pathologies. The host immune response to hbv antigens is a critical factor determining the outcome of the infection. In this thesis we have developed an immune-gene therapy strategy for the treatment of chb. As gene vehicle, we employed recombinant aav viruses and the expression of two immunostimulatory citokines, ifn-α and il-15. In the first part, we tested the functionality of both virus in vivo and determine the optimal dosage. Next, we tested the antiviral activity of both vectors in a highly tolerant animal model of chb, hbv transgenic mice. we found that aavil-15 therapy increased serum levels of il-15 bound to il-15rα leading to robust intrahepatic expansion of ifn-gamma-producing lymphocytes and increased hbv-specific cd8+ t-lymphocytes which, however, lacked cytolytic activity. Contrariwise, aavifn-α stimulated cytotoxic functions of hbv-specific cd8+t-cells without increasing their number. Notably, administration of aavil-15+aavifn-α to transgenic mice increased both the number and the cytolytic activity of anti-hbv cd8+t cells within the liver causing extensive hepatocellular necrosis and viral clearance. Aavil-15-treated transgenic mice showed ifn-gamma-dependent hepatic pd-l1 upregulation and a high proportion of pd-1+ intrahepatic t-cells. Combining aavifn-α with aavil-15 reduced pd-1 and pd-l1 expression allowing hbv-specific t-lymphocytes to kill target cells. Accordingly, treatment of hbv transgenic mice with aavil-15 plus anti-pd-l1 antibody caused intense cytolysis and viral clearance. Our study defines a potent new approach to break immune tolerance with broad applications in chronic viral infections and neoplastic conditions. abbreviation: aav: adeno-associated virus; chb: chronic hepatitis b; ifn-α: interferon-alpha; il-15: interleukin-15; hbv: human hepatitis b virus; pd-1: programmed-death 1; pd-l1: programmed-death ligand 1.
Datos académicos de la tesis doctoral «Gene transfer of immunomodulatory cytokines: biological and functional characterization of adenoassociated virus expressing interferon-alpha and interleukin-15 in a hbv transgenic mouse model.«
- Título de la tesis: Gene transfer of immunomodulatory cytokines: biological and functional characterization of adenoassociated virus expressing interferon-alpha and interleukin-15 in a hbv transgenic mouse model.
- Autor: María nna Di Scala
- Universidad: Navarra
- Fecha de lectura de la tesis: 21/06/2013
Dirección y tribunal
- Director de la tesis
- Gloria Gonzales Aseguinolaza
- Tribunal
- Presidente del tribunal: Jesús m. Prieto valtueña
- José Carlos Segovia sanz (vocal)
- jessica Fioravanti (vocal)
- Luis Alberto Anel bernal (vocal)
