Skip to main content
Login | Suomeksi | På svenska | In English

Generation and characterization of a novel oncolytic adenovirus expressing OX40L and CD40L

Show simple item record 2020-06-05T10:20:42Z 2020-06-05T10:20:42Z 2020-06-05
dc.title Generation and characterization of a novel oncolytic adenovirus expressing OX40L and CD40L en
ethesis.discipline biotekniikka fi
ethesis.discipline Biotechnology en
ethesis.discipline bioteknik sv
ethesis.discipline.URI und
ethesis.department Bio- ja ympäristötieteellinen tiedekunta fi
ethesis.department Faculty of Biological and Environmental Sciences en
ethesis.department Bio- och miljövetenskapliga fakulteten sv
ethesis.faculty Bio- ja ympäristötieteellinen tiedekunta fi
ethesis.faculty Faculty of Biological and Environmental Sciences en
ethesis.faculty Bio- och miljövetenskapliga fakulteten sv
ethesis.faculty.URI Helsingin yliopisto fi University of Helsinki en Helsingfors universitet sv
dct.creator Martins, Beatriz
dct.issued 2020
dct.language.ISO639-2 eng
dct.abstract According to the latest estimations, cancer is the second leading cause of death worldwide. Despite the significant advances in the range of drugs and treatment modalities to treat cancer, the number of deaths is estimated to continue rising, posing serious challenges for the patients, their families, and the healthcare systems. Conventional treatments tend to be associated with severe adverse side effects and treatment resistance. Consequently, safer and more efficient therapy options are urgently needed, especially for the treatment of metastatic tumors refractory to conventional treatments. A new and revolutionizing field in oncology is immunotherapy, in which oncolytic viruses are included. Oncolytic viruses have an inherent or acquired selectivity to replicate exclusively in tumor cells, ultimately destroying them. Simultaneously, they also activate the dormant host’s immune system to fight against the tumor. Adenoviruses, particularly, have shown to be safe, inducing only mild adverse side effects in clinical trials, making them a great candidate for further clinical development. Adenoviruses can be genetically modified to increase their infectivity or improve the anti-cancer immune responses induced by the virus, e.g., through the expression of immunostimulatory molecules. The focus of this thesis was to develop and characterize several genetically modified oncolytic adenoviruses expressing either OX40L alone or OX40L and CD40L, two co-stimulatory molecules capable of engaging both the innate and adaptive arms of the immune system to fight the tumor. The insertion of the transgenes into the E3B-14.7k region of the Ad5/3-∆24 adenovector plasmid was performed using Gibson Assembly® cloning approach. After successful cloning, the recombinant viral genomes were transfected into A549 cells for viral amplification, followed by CsCl purification to produce a high titer viral preparation. The expression of the transgenes was studied in vitro by ELISA and functional assays, showing promising expression levels of functional OX40L and CD40L. However, when the infectivity and virus killing potency were analyzed, in vitro by immunocytochemistry and MTS assay; and in vivo using an immunodeficient mouse model, the data showed that the cloned viruses performed sub-optimally when compared to the control unarmed virus (Ad5/3-∆24). These findings suggest that the insertion of the two transgenes in place of the E3-14.7k gene was detrimental to the fitness of the virus. en
dct.subject cancer en
dct.subject oncolytic adenovirus en
dct.subject OX40L en
dct.subject CD40L en
dct.subject Gibson Assembly en
dct.language en
ethesis.language English en
ethesis.language englanti fi
ethesis.language engelska sv
ethesis.supervisor Cerullo, Vincenzo
ethesis.supervisor Ylösmäki, Erkko
ethesis.thesistype pro gradu -tutkielmat fi
ethesis.thesistype master's thesis en
ethesis.thesistype pro gradu-avhandlingar sv
dct.identifier.ethesis E-thesisID:fc586ef5-1551-4d96-a7bd-92e1ef3eb02e
ethesis-internal.timestamp.reviewStep 2020-04-21 10:34:42:716
ethesis.principalprofessor Keinänen, Kari
dct.identifier.urn URN:NBN:fi:hulib-202006052630
dc.type.dcmitype Text
ethesis.facultystudyline.URI none und
ethesis.mastersdegreeprogram.URI none und

Files in this item

Files Size Format View
Martins_Beatriz_pro_gradu_2020.pdf 18.36Mb PDF

This item appears in the following Collection(s)

Show simple item record