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dc.contributor.authorSubbanna, Sowmya
dc.date.accessioned2018-05-10T22:00:03Z
dc.date.issued2018
dc.identifier.citationSubbanna, Sowmya. The microenvironment as the basis of CLL cell growth - Defining pathways, enabling EBV transformation and performing groundwork for drug sensitivity screening of patients. Master thesis, University of Oslo, 2018
dc.identifier.urihttp://hdl.handle.net/10852/61660
dc.description.abstractChronic lymphocytic leukemia (CLL) is a mature B-cell malignancy that most often affects the elderly. CLL cells depend on their microenvironment for their extended growth and survival. CLL cells are difficult to grow “in vitro” as they undergo cell death in cultures. Moreover, it has been difficult to transform CLL cells with EBV a common technique otherwise used to generate lymphoblastic cell lines (LCL) from primary B-cells. These features have made study of disease pathogenesis more difficult. Here, I present experiments that allow long term culture of CLL cells and I tested whether this culture method could support EBV transformation of CLL cells. Moreover, the thesis presents experiments where the in vitro drug sensibility of CLL cells is tested. In contrast to normal B-cells, CLL cells do not regularly become activated and immortalized after exposure to EBV, and the virus does not induce CLL cell proliferation (1, 2). This phenomenon may be linked to the cell cycle block of CLL cells. To overcome this issue, we stimulated CLL cells with CD40L+BAFF+ L cells in the presence of cytokines and added EBV. We found that CD40L+BAFF+ L cells and IL-2 supported the transformation and survival of EBV-EGFP+ CLL cells and IL-2 supported the transformation and survival of EBV-EGFP+ CLL cells for 3 months. However, results varied between patients. Further, we also investigated the CLL microenvironmental factors (mainly L cells and cytokines) that are involved in supporting the CLL cell growth, proliferation and survival. We observed that CD40L+BAFF+ L cells were potent and supported CLL cells proliferation and blastogenesis. CD40L+BAFF+ L cells and IL-2 or IL-21 also showed a significant increase in CLL cell proliferation and blastogenesis respectively. In addition, we observed that CD40L+APRIL+ L cells or CD40L+BAFF+L cells supplemented with IL-15 or IL-6 enhanced CLL cell proliferation and blastogenesis. We also found that mixture of L cells (CD40L+APRIL+BAFF+) co-cultured together along with cytokines also enhanced CLL cell proliferation and blasts to higher extent. We also investigated the effects of the cytostatic drugs Fludarabine, Doxorubicine and Vincristine on CLL cells. Doxorubicin was more effective even with 1µM concentration of the drug. Scaling up such in vitro testing should allow drug sensitivity screens of cells from patients and suggests a strategy that may be helpful for personalized medicine. In conclusion, stimulation and ex vivo culture of CLL cells may aid the understanding of important microenvironmental factors that cause CLL cell growth, may give information required for EBV transformation of CLL cells and may be the key for successful immortalization of CLL cells and allow for discovery of patient specific drug sensitivities.eng
dc.language.isoeng
dc.subject
dc.titleThe microenvironment as the basis of CLL cell growth - Defining pathways, enabling EBV transformation and performing groundwork for drug sensitivity screening of patientseng
dc.typeMaster thesis
dc.date.updated2018-05-10T22:00:02Z
dc.creator.authorSubbanna, Sowmya
dc.date.embargoenddate3018-01-25
dc.rights.termsDette dokumentet er ikke elektronisk tilgjengelig etter ønske fra forfatter. Tilgangskode/Access code A
dc.identifier.urnURN:NBN:no-64266
dc.type.documentMasteroppgave
dc.rights.accessrightsclosedaccess
dc.identifier.fulltextFulltext https://www.duo.uio.no/bitstream/handle/10852/61660/1/Master-Thesis_Sowmya-Subbanna.pdf


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