Strategies for improved cancer virotherapy [Elektronische Ressource] : in vivo migration and expansion of dendritic cells enhance cross-presentation of tumor antigens in virotherapy / von Edukulla Ramakrishna
Strategies for Improved Cancer Virotherapy: In vivo migration and expansion of dendritic cells enhance cross-presentation of tumor antigens in virotherapy Von der Naturwissenschaftlichen Fakultät der Gottfried Wilhelm Leibniz Universität Hannover zur Erlangung des akademischen Grades Doktor der Naturwissenschaften Dr. rer. nat. genehmigte dissertation von Edukulla Ramakrishna Master of Science in Biochemistry geboren am 05.03.1975 in Chandoor, Indien Hannover 2008 This work is dedicated to the goddess of study and my family members Die vorliegende Arbeit wurde in der Abteilung Gastroenterologie, Endokrinologie and Hepatologie der Medizinischen Hochschule Hannover in der Zeit vom 20.09.2004 bis zum 22.11.2007 unter der Leitung von Prof. Dr.med. Stefan Kubicka angefertigt. Referent Prof. Bernd Otto Institut für Physiologische Chemie Tierärztliche Hochschule Hannover Koreferent Prof. Walter Müller für Biochemie Medizinische Hochschule Hannover Tag der Promotion 17.01.2008 Contents Contents 1 Abstract..................................................................................................................1 2 Zusammenfassung..................................................................................................2 3 Introduction..................
Von der Naturwissenschaftlichen Fakultät der Gottfried Wilhelm Leibniz Universität Hannover zur Erlangung des akademischen Grades Doktor der Naturwissenschaften Dr. rer. nat. genehmigte dissertation von Edukulla Ramakrishna Master of Science in Biochemistry geboren am 05.03.1975 in Chandoor, Indien
Hannover 2008
This work is dedicated to the goddess of study and my family members
Die vorliegende Arbeit wurde in der Abteilung Gastroenterologie, Endokrinologie and
Hepatologie der Medizinischen Hochschule Hannover in der Zeit vom 20.09.2004 bis
zum 22.11.2007 unter der Leitung von Prof. Dr.med. Stefan Kubicka angefertigt.
Viroimmunotherapy expands cytotoxic T cellsin vivoand kills antigen-
specific target cells...................................................................................................51
6.9In vivomaturation of DCs leads to migration of DCs to lymph nodes to activate naïve T cells................................................................................................54
6.10 Viroimmunotherapyand subsequent tumor antigen loaded DC vaccination
inhibit established lung metastasis...........................................................................56
6.11 Combination of oncolytic virotherapy and immunotherapy increases spleen
size and eradicates established subcutaneous tumors ..............................................59
Triggering of dendritic cell (DC) migration into tumor tissue and subsequent
expansion of dendritic cells may be useful to generate anti-tumor immune response by
adenoviral gene transfer employing cytokines and growth factors for dendritic cell
migration and expansion to improve cross-presentation of tumor antigens during
oncolytic virotherapy.
Appropriate syngenic mouse models were established for lung carcinoma cells
CMT64 and KLN205.In vitroreplication assay showed that CMT64 and KLN205 are
permissive for replication of the oncolytic telomerase-dependent adenovirus hTERT-
Ad. Furtherin vivo replication was studied using replication defective adenovirus
expressing Luciferase gene in combination of telomerase dependent adenovirus.
We used adenoviral expression of Macrophage Inflammatory Protein 1α(MIP-1α) to
recruit DCs into hTERT-Ad infected tumors and Ad-Flt3L to expand DCs within the
tumor nodules during virotherapy.
Compared with virotherapy alone or virotherapy with expression of a single CC
chemokine, expression of both MIP1α and Flt3L during virotherapy significantly
increases infiltration of DCs and T cells, as shown by histochemistry. Interferon-γ,
and CTL assay showed that the combination of cytokines improve cross-presentation
of tumor antigens and induces an anti-tumor immune response during virotherapy.
Combination of viroimmunotherapy and DC vaccination showed nearly complete
regression of the subcutaneous tumors and very strong inhibition of established lung
metastasis.
In conclusion mobilizing of DCs to tumors during virotherapy improves cross-
presentation of tumor antigens and induced anti-tumor immune response, which
facilitates an effective viroimmunotherapy of established solid tumorsin vivo. The results show high efficacy of the viroimmunotherapy, which may be considered as
safe even, when translated to human cancer therapy.