5763
2011
eng
article
1
2012-04-13
--
--
Specific antibody-receptor interactions trigger InlAB-independent uptake of Listeria monocytogenes into tumor cell lines
Background: Specific cell targeting is an important, yet unsolved problem in bacteria-based therapeutic applications, like tumor or gene therapy. Here, we describe the construction of a novel, internalin A and B (InlAB)-deficient Listeria monocytogenes strain (Lm-spa+), which expresses protein A of Staphylococcus aureus (SPA) and anchors SPA in the correct orientation on the bacterial cell surface. Results: This listerial strain efficiently binds antibodies allowing specific interaction of the bacterium with the target recognized by the antibody. Binding of Trastuzumab (Herceptin®) or Cetuximab (Erbitux®) to Lm-spa+, two clinically approved monoclonal antibodies directed against HER2/neu and EGFR/HER1, respectively, triggers InlABindependent internalization into non-phagocytic cancer cell lines overexpressing the respective receptors. Internalization, subsequent escape into the host cell cytosol and intracellular replication of these bacteria are as efficient as of the corresponding InlAB-positive, SPA-negative parental strain. This specific antibody/receptormediated internalization of Lm-spa+ is shown in the murine 4T1 tumor cell line, the isogenic 4T1-HER2 cell line as well as the human cancer cell lines SK-BR-3 and SK-OV-3. Importantly, this targeting approach is applicable in a xenograft mouse tumor model after crosslinking the antibody to SPA on the listerial cell surface. Conclusions: Binding of receptor-specific antibodies to SPA-expressing L. monocytogenes may represent a promising approach to target L. monocytogenes to host cells expressing specific receptors triggering internalization.
urn:nbn:de:bvb:20-opus-68705
6870
BMC Microbiology (2011) 11:163, doi:10.1186/1471-2180-11-163
Martin Heisig
Alexa Frentzen
Birgit Bergmann
Katharina Ivaylo Gentschev
Christian Hotz
Christoph Schoen
Jochen Stritzker
Joachim Fensterle
Ulf R. Rapp
Werner Goebel
deu
swd
Listeria monocytogenes
Medizin und Gesundheit
open_access
Institut für Hygiene und Mikrobiologie
Förderzeitraum 2011
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/5763/Heisig_1471_2180_11_163.pdf
12570
2015
eng
4075-4092
7
article
1
2016-01-27
--
--
Preclinical Testing Oncolytic Vaccinia Virus Strain GLV-5b451 Expressing an Anti-VEGF Single-Chain Antibody for Canine Cancer Therapy
Virotherapy on the basis of oncolytic vaccinia virus (VACV) strains is a novel approach for canine cancer therapy. Here we describe, for the first time, the characterization and the use of VACV strain GLV-5b451 expressing the anti-vascular endothelial growth factor (VEGF) single-chain antibody (scAb) GLAF-2 as therapeutic agent against different canine cancers. Cell culture data demonstrated that GLV-5b451 efficiently infected and destroyed all four tested canine cancer cell lines including: mammary carcinoma (MTH52c), mammary adenoma (ZMTH3), prostate carcinoma (CT1258), and soft tissue sarcoma (STSA-1). The GLV-5b451 virus-mediated production of GLAF-2 antibody was observed in all four cancer cell lines. In addition, this antibody specifically recognized canine VEGF. Finally, in canine soft tissue sarcoma (CSTS) xenografted mice, a single systemic administration of GLV-5b451 was found to be safe and led to anti-tumor effects resulting in the significant reduction and substantial long-term inhibition of tumor growth. A CD31-based immuno-staining showed significantly decreased neo-angiogenesis in GLV-5b451-treated tumors compared to the controls. In summary, these findings indicate that GLV-5b451 has potential for use as a therapeutic agent in the treatment of CSTS.
Viruses
10.3390/v7072811
urn:nbn:de:bvb:20-opus-125705
Viruses 2015, 7, 4075-4092; doi:10.3390/v7072811
Marion Adelfinger
Simon Bessler
Alexander Cecil
Johanna Langbein-Laugwitz
Alexa Frentzen
Ivaylo Gentschev
Aladar A. Szalay
eng
uncontrolled
canine cancer therapy
eng
uncontrolled
canine soft tissue sarcoma (CSTS)
eng
uncontrolled
oncolytic virus
eng
uncontrolled
cancer
eng
uncontrolled
canine cancer cell lines
eng
uncontrolled
antibody production
eng
uncontrolled
angiogenesis
Medizin und Gesundheit
open_access
Julius-von-Sachs-Institut für Biowissenschaften
Lehrstuhl für Biochemie
Förderzeitraum 2015
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/12570/Szalay_viruses-07-02811.pdf
11938
2014
eng
e104337
8
9
article
1
2015-09-29
--
--
Evaluation of a New Recombinant Oncolytic Vaccinia Virus Strain GLV-5b451 for Feline Mammary Carcinoma Therapy
Virotherapy on the basis of oncolytic vaccinia virus (VACV) infection is a promising approach for cancer therapy. In this study we describe the establishment of a new preclinical model of feline mammary carcinoma (FMC) using a recently established cancer cell line, DT09/06. In addition, we evaluated a recombinant vaccinia virus strain, GLV-5b451, expressing the anti-vascular endothelial growth factor (VEGF) single-chain antibody (scAb) GLAF-2 as an oncolytic agent against FMC. Cell culture data demonstrate that GLV-5b451 virus efficiently infected, replicated in and destroyed DT09/06 cancer cells. In the selected xenografts of FMC, a single systemic administration of GLV-5b451 led to significant inhibition of tumor growth in comparison to untreated tumor-bearing mice. Furthermore, tumor-specific virus infection led to overproduction of functional scAb GLAF-2, which caused drastic reduction of intratumoral VEGF levels and inhibition of angiogenesis.
In summary, here we have shown, for the first time, that the vaccinia virus strains and especially GLV-5b451 have great potential for effective treatment of FMC in animal model.
PLoS ONE
10.1371/journal.pone.0104337
urn:nbn:de:bvb:20-opus-119387
PLoS ONE 9(8): e104337. doi:10.1371/journal.pone.0104337
Marion Adelfinger
Ivaylo Gentschev
Julio Grimm de Guibert
Stephanie Weibel
Johanna Langbein-Laugwitz
Barbara Härtl
Hugo Murua Escobar
Ingo Nolte
Nanhai G. Chen
Richard J. Aguilar
Yong A. Yu
Qian Zhang
Alexa Frentzen
Aladar A. Szalay
eng
uncontrolled
antibodies
eng
uncontrolled
cancer treatment
eng
uncontrolled
carcinomas
eng
uncontrolled
vaccinia virus
eng
uncontrolled
oncolytic viruses
eng
uncontrolled
viral replication
eng
uncontrolled
cell cultures
eng
uncontrolled
enzyme-linked immunoassays
Krankheiten
open_access
Institut für Molekulare Infektionsbiologie
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/11938/033_Adelfinger_PLOS_ONE.pdf
2193
2007
deu
doctoralthesis
1
2007-12-21
--
2007-12-19
Posttranskriptionale Regulation der Internalinexpression und alternative Internalin-unabhängige Aufnahme von Listeria monocytogenes in Animalzellen
Posttranscriptional regulation of the Internalin Expression and Alternative Internalin Independent Uptake of Listeria monocytogenes in Animal Cells
Listeria monocytogenes ist ein weit verbreitetes, Gram-positives humanpatho-genes Bakterium, welches in immunsupprimierten Personen das Krankheitsbild der Listeriose auslösen kann. Der Infektionszyklus der Listerien im Wirt ist im Hinblick auf die Pathogenese dieses Erregers intensiv untersucht worden. Die Regulation der verschiedenen beteiligten Virulenzfaktoren unterliegt in L. monocytogenes einer starken Kontrolle, die einerseits durch regulatorische Proteine aber auch durch Umweltfaktoren beeinflusst wird. Die Mechanismen, die auf transkriptionaler wie auch auf translationaler Ebene die Expression verschiedener listerieller Virulenzgene regulieren, wurden kürzlich näher charakterisiert. Es wurden für verschiedene listerielle Virulenzgene Riboswitch-mechanismen zur Expressionskontrolle in Listerien neu beschrieben. Durch Vorarbeiten wurde auch für das inlAB-Operon ein posttranskriptionaler Regu-lationsmechanismus postuliert. Dabei wurde der anaerobe Stoffwechsel der Listerien als möglicher Auslöser für die beobachtete Translationssteigerung des inlA- und inlB-Gens diskutiert. Innerhalb der vorliegenden Arbeit sollte nun weitergehend untersucht werden, in welchem Bereich der Sequenz des inlAB-Operons sich regulatorische Strukturen zur posttranskriptionalen Regulation unter anaeroben Wachstumsbedingungen befinden. Dazu wurden verschiedene Mutanten mit unterschiedlichen Deletionen im inlAB-Operon konstruiert und die Transkription und Translation sowohl des inlA-, als auch des inlB-Gens betrachtet. Eine Deletion im aroA-Gen bewirkt das Wachstum der Bakterien bei anaerobem Stoffwechsel. Diese Deletion wurde in die konstruierten Stämme eingefügt, um die Expression der Gene unter den verschiedenen Wachstumsbedingungen vergleichen zu können. Außerdem wurden verschiedene gus-Reportergen-Fusionsmutanten und Promotor-austauschmutanten konstruiert, um quantitativ aussagekräftigere Daten zu erheben. Die Charakterisierung der Mutanten ließ erkennen, dass keiner der deletierten Bereiche des inlAB-Operons von L. monocytogenes für die beobachtete Translationssteigerung im inlA-Gen bei anaerobem Stoffwechsel verantwortlich zu sein scheint. Das inlB-Gen war innerhalb der hier gezeigten Experimente nicht posttranskriptional reguliert, wie im Vorfeld postuliert. Nach plasmidkodierter Expression verschiedener Bereiche des inlAB-Operons konnte, verglichen mit genomischer Expression, keine Veränderung in der inlA-Expression beobachtet werden. Die mögliche Beteiligung eines potentiellen Regulatorproteins konnte innerhalb dieser Arbeit daher nicht näher eingegrenzt werden. Auch ein Einfluss der regulatorischen Faktoren Hfq und CcpA auf die Expression des InlA Proteins in der L. monocytogenes ΔaroA-Mutante konnte nicht gefunden werden. Es zeigte sich interessanterweise außerdem, dass weitere Virulenzgene wie actA und hly unter den anaeroben Bedingungen ebenfalls eine Translations-steigerung zeigten. Somit stellt sich abschließend die Frage, ob es sich bei der beobachteten Translationssteigerung des inlA-Gens wirklich um einen durch bestimmte Strukturen in der inlAB-mRNA ausgelösten Mechanismus handelt. L. monocytogenes ist als intrazellulär replizierendes, Gram-positives Bakterium interessant für den Einsatz in immun- und tumortherapeutischen Anwendungen. Attenuierte L. monocytogenes-Stämme wurden dazu bereits erfolgreich im Mausmodell als Trägerbakterien für Impfstoffstrategien eingesetzt. Die gezielte Infektion von Geweben ist jedoch aufgrund des wenig ausgeprägten Zelltropismus der Listerien im Wirt bisher ein Problem für einen Einsatz in bakterienbasierten Anwendungen, wie z.B. der Tumor- oder Gentherapie. Innerhalb dieser Arbeit wurden L. monocytogenes-Stämme konstruiert, bei denen chromosomal das für die Integrase codierende Gen gegen das Gen für das Staphylokokken Protein A (SPA) unter der Kontrolle listerieller Promotoren ausgetauscht wurde. Die erfolgreiche Oberflächenlokalisation von Protein A in der Zellwand von Listerien konnte im Western Blot oder in funktionellen Immunfluoreszenzfärbungen in Mikroskop- und FACS-Analysen nachgewiesen werden. Diese Stämme sollen im Cell Targeting zur gezielten Infektion von Geweben eingesetzt werden. Dazu konnten die Bakterien über Herceptin®-HER2/neu-vermittelte Adhäsion an SK-BR-3-Zellen erfolgreich in diese aufgenommen werden und innerhalb dieser replizieren. Die neu konstruierten Listeria-Stämme zeigten im Mausmodell keine Veränderung in ihrer Virulenz verglichen mit nicht-SPA-exprimierenden Stämmen. Die in dieser Arbeit vorgestellte Antikörper-Rezeptor-vermittlelte Aufnahme der Listerien in Zellen stellt einen neuen, bisher nicht beschriebenen Mechanismus dar, der in vielen therapeutischen Anwendungen zur Infektion spezifischer Gewebe durch Listerien genutzt werden kann.
Listeria monocytogenes is a widely distributed gram-positive pathogenic bacterium, that causes the disease listeriosis in immunocompromised persons. The infectious cycle of this pathogen has been well investigated concerning the mechanisms of pathogenesis. The regulation of the involved virulence factors underlie strong control mechanisms both by regulatory proteins and environmental conditions. The mechanisms that control the expression of listerial virulence genes on the transcriptional and translational level have recently been characterized more closely. For some of these virulence factors, regulatory posttranscriptional riboswitch control mechanisms have recently been newly described in Listeria. In studies prior to this work, a posttranscriptional regulation mechanism has been postulated for the inlAB-Operon, also. As a trigger for the enhanced translation of the inlA and inlB gene the anaerobic metabolism of Listeria has been discussed. In the here presented work, it should be further investigated which part of the inlAB operon comprises regulatory structures for the posttranscriptional regulation at anaerobic growth conditions. To this purpose, different mutants with varying deletions in the inlAB operon have been constructed and transcription and translation of both the inlA and inlB genes observed. A deletion in the aroA gene provokes growth of the bacteria at anaerobic metabolism. This deletion was introduced in the newly constructed strains to be able to compare the different growth conditions. Additionally, gus-reportergene fusion as well as promoter exchange mutants were constructed to be able to raise quantitatively significant data. The characterization of the deletion mutants showed that none of the introduced deletions in the inlAB operon seems to be responsible for the enhanced expression of inlA in anaerobically growing Listeriae. In the here presented experiments, the inlB-gene was not found to be posttranscriptionally regulated, in accordance with a previous report. The plasmid encoded expression of different parts of the inlAB operon did not lead to an alteration in the inlA expression compared to genomically encoded inlA. The possible interaction of a potential regulator protein could thus not be further elucidated. Also, the engagement of the regulators like Hfq or CcpA could not be found. Interestingly, also the virulence genes actA and hly showed an enhanced translational efficiency under the given conditions. Thus, in the herein presented work, the question is raised, whether the observed translation enhancement of the inlA gene is really due to regulatory structures within the inlAB-mRNA. As an intracellularly replicating, Gram positive bacterium, L. monocytogenes is a very interesting means to be used in immuno- or tumortherapeutic applications. Attenuated L. monocytogenes strains have already been successfully applied as carrier strains in vaccine strategies in the mouse model. Due to the distinct cell tropism of Listeriae in the host, the targeted infection of tissue has so far been a problem regarding the use in bacteria-based applications like tumor or gene therapy. In this work, L. monocytogenes strains have been constructed, that contain the chromosomal integrase gene exchanged for the staphylococcal Protein A (SPA) gene under the control of different listerial promoters. The successful translocation and anchoring of SPA in the cell wall was confirmed by either western blot or functionally by immunofluorescence staining using microscopic and FACS analysis. These strains should be used to specifically infect tissues via cell targeting. Through Herceptin®-HER2/neu-mediated adhesion to SK-BR-3 cells the bacteria could be successfully taken up and replicate in these cells. The newly constructed Listeria strains did not show any alteration in virulence in the mouse model when compared to non-SPA-expressing strains. The herein presented antibody-receptor-mediated uptake of the Listeriae in cells demonstrates a new, so far not described mechanism, that can be used in many therapeutic applications involving the infection of specific tissues by Listerae.
urn:nbn:de:bvb:20-opus-25631
2563
X121643
Alexa Frentzen
deu
swd
Listeria monocytogenes
deu
swd
Internalin
deu
swd
Genregulation
deu
swd
Aufnahme
eng
uncontrolled
Listeria monocytogenes
eng
uncontrolled
Internalin
eng
uncontrolled
Gene Regulation
eng
uncontrolled
Uptake
Biowissenschaften; Biologie
open_access
Theodor-Boveri-Institut für Biowissenschaften
Universität Würzburg
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/2193/Dissertation_Alexa_Frentzen_Juni_2007.pdf
13003
2012
eng
e47472
10
7
article
1
2016-03-16
--
--
Virotherapy of Canine Tumors with Oncolytic Vaccinia Virus GLV-1h109 Expressing an Anti-VEGF Single-Chain Antibody
Virotherapy using oncolytic vaccinia virus (VACV) strains is one promising new strategy for cancer therapy. We have previously reported that oncolytic vaccinia virus strains expressing an anti-VEGF (Vascular Endothelial Growth Factor) single-chain antibody (scAb) GLAF-1 exhibited significant therapeutic efficacy for treatment of human tumor xenografts. Here, we describe the use of oncolytic vaccinia virus GLV-1h109 encoding GLAF-1 for canine cancer therapy. In this study we analyzed the virus-mediated delivery and production of scAb GLAF-1 and the oncolytic and immunological effects of the GLV-1h109 vaccinia virus strain against canine soft tissue sarcoma and canine prostate carcinoma in xenograft models. Cell culture data demonstrated that the GLV-1h109 virus efficiently infect, replicate in and destroy both tested canine cancer cell lines. In addition, successful expression of GLAF-1 was demonstrated in virus-infected canine cancer cells and the antibody specifically recognized canine VEGF. In two different xenograft models, the systemic administration of the GLV-1h109 virus was found to be safe and led to anti-tumor and immunological effects resulting in the significant reduction of tumor growth in comparison to untreated control mice. Furthermore, tumor-specific virus infection led to a continued production of functional scAb GLAF-1, resulting in inhibition of angiogenesis. Overall, the GLV-1h109-mediated cancer therapy and production of immunotherapeutic anti-VEGF scAb may open the way for combination therapy concept i.e. vaccinia virus mediated oncolysis and intratumoral production of therapeutic drugs in canine cancer patients.
PLoS One
10.1371/journal.pone.0047472
urn:nbn:de:bvb:20-opus-130039
PLoS ONE 7(10): e47472. doi:10.1371/journal.pone.0047472
Sandeep S. Patil
Ivaylo Gentschev
Marion Adelfinger
Ulrike Donat
Michael Hess
Stephanie Weibel
Ingo Nolte
Alexa Frentzen
Aladar A. Szalay
eng
uncontrolled
angiogenesis
eng
uncontrolled
microenvironment
eng
uncontrolled
model
eng
uncontrolled
cancer
eng
uncontrolled
therapy
eng
uncontrolled
pet dogs
eng
uncontrolled
nude-mice
eng
uncontrolled
breast-tumors
eng
uncontrolled
microvascular density
eng
uncontrolled
endothelial growth-factor
Medizin und Gesundheit
open_access
Rudolf-Virchow-Zentrum
Lehrstuhl für Biochemie
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/13003/journal.pone.0047472.pdf
9601
2013
eng
article
1
--
--
--
Treatment of malignant effusion by oncolytic virotherapy in an experimental subcutaneous xenograft model of lung cancer
Background
Malignant pleural effusion (MPE) is associated with advanced stages of lung cancer and is mainly dependent on invasion of the pleura and expression of vascular endothelial growth factor (VEGF) by cancer cells. As MPE indicates an incurable disease with limited palliative treatment options and poor outcome, there is an urgent need for new and efficient treatment options.
Methods
In this study, we used subcutaneously generated PC14PE6 lung adenocarcinoma xenografts in athymic mice that developed subcutaneous malignant effusions (ME) which mimic pleural effusions of the orthotopic model. Using this approach monitoring of therapeutic intervention was facilitated by direct observation of subcutaneous ME formation without the need of sacrificing mice or special imaging equipment as in case of MPE. Further, we tested oncolytic virotherapy using Vaccinia virus as a novel treatment modality against ME in this subcutaneous PC14PE6 xenograft model of advanced lung adenocarcinoma.
Results
We demonstrated significant therapeutic efficacy of Vaccinia virus treatment of both advanced lung adenocarcinoma and tumor-associated ME. We attribute the efficacy to the virus-mediated reduction of tumor cell-derived VEGF levels in tumors, decreased invasion of tumor cells into the peritumoral tissue, and to viral infection of the blood vessel-invading tumor cells. Moreover, we showed that the use of oncolytic Vaccinia virus encoding for a single-chain antibody (scAb) against VEGF (GLAF-1) significantly enhanced mono-therapy of oncolytic treatment.
Conclusions
Here, we demonstrate for the first time that oncolytic virotherapy using tumor-specific Vaccinia virus represents a novel and promising treatment modality for therapy of ME associated with advanced lung cancer.
Journal of Translational Medicine
doi:10.1186/1479-5876-11-106
http://www.translational-medicine.com/content/11/1/106
urn:nbn:de:bvb:20-opus-96016
MRB Forschungszentrum für Magnet-Resonanz-Bayern e.V., Am Hubland, D-97074 Würzburg
Genelux Corporation, San Diego Science Center, 3030 Bunker Hill Street, Suite 310, San Diego, California 92109, USA
In: Journal of Translational Medicine (2013) 11: 106, doi:10.1186/1479-5876-11-106
Aladar A Szalay
Stephanie Weibel
Elisabeth Hofmann
Thomas Christian Basse-Luesebrink
Ulrike Donat
Carolin Seubert
Marion Adelfinger
Prisca Gnamlin
Christina Kober
Alexa Frentzen
Ivaylo Gentschev
Peter Michael Jakob
eng
uncontrolled
Oncolytic virotherapy
eng
uncontrolled
Malignant effusion
eng
uncontrolled
Lung cancer
eng
uncontrolled
VEGF
deu
swd
Lungenkrebs
deu
swd
Vascular endothelial Growth Factor
Medizin und Gesundheit
open_access
Physikalisches Institut
Institut für Molekulare Infektionsbiologie
Rudolf-Virchow-Zentrum
Lehrstuhl für Biochemie
Förderzeitraum 2013
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/9601/Szalay_1479-5876-11-106.pdf
17078
2017
eng
41-61
5
article
1
2018-10-30
--
--
Humanized Mice with Subcutaneous Human Solid Tumors for Immune Response Analysis of Vaccinia Virus-Mediated Oncolysis
Oncolytic vaccinia virus (VACV) therapy is an alternative cancer treatment modality that mediates targeted tumor destruction through a tumor-selective replication and an induction of anti-tumor immunity. We developed a humanized tumor mouse model with subcutaneous human tumors to analyze the interactions of VACV with the developing tumors and human immune system. A successful systemic reconstitution with human immune cells including functional T cells as well as development of tumors infiltrated with human T and natural killer (NK) cells was observed. We also demonstrated successful in vivo colonization of such tumors with systemically administered VACVs. Further, a new recombinant GLV-1h376 VACV encoding for a secreted human CTLA4-blocking single-chain antibody (CTLA4 scAb) was tested. Surprisingly, although proving CTLA4 scAb’s in vitro binding ability and functionality in cell culture, beside the significant increase of CD56\(^{bright}\) NK cell subset, GLV-1h376 was not able to increase cytotoxic T or overall NK cell levels at the tumor site. Importantly, the virus-encoded β-glucuronidase as a measure of viral titer and CTLA4 scAb amount was demonstrated. Therefore, studies in our “patient-like” humanized tumor mouse model allow the exploration of newly designed therapy strategies considering the complex relationships between the developing tumor, the oncolytic virus, and the human immune system.
Molecular Therapy Oncolytics
10.1016/j.omto.2017.03.001
28480327
urn:nbn:de:bvb:20-opus-170786
Molecular Therapy Oncolytics 2017, Vol. 5, 41-61. DOI: 10.1016/j.omto.2017.03.001
false
true
CC BY-NC-ND: Creative-Commons-Lizenz: Namensnennung, Nicht kommerziell, Keine Bearbeitungen 4.0 International
Desislava Tsoneva
Boris Minev
Alexa Frentzen
Qian Zhang
Anja K. Wege
Aladar A. Szalay
eng
uncontrolled
humanized tumor
eng
uncontrolled
mouse model
eng
uncontrolled
subcutaneous human tumors
eng
uncontrolled
Oncolytic vaccinia virus
Biochemie
open_access
Rudolf-Virchow-Zentrum
Lehrstuhl für Biochemie
Universität Würzburg
https://opus.bibliothek.uni-wuerzburg.de/files/17078/064_Tsoneva_MOLECULAR-THERAPY-ONCOLYTICS.pdf