Antitumor virotherapy using syngeneic or allogeneic mesenchymal stem cell carriers induces systemic immune response and intratumoral leukocyte infiltration in mice
Oncolytic virotherapy uses oncolytic viruses that selectively replicate in cancer cells. The use of cellular vehicles with migration ability to tumors has been considered to increase their delivery to target sites. Following this approach, the antitumor efficacy of the treatment Celyvir (mesenchymal stem cells infected with the oncolytic adenovirus ICOVIR-5) has been demonstrated in patients with neuroblastoma. However, the better efficacy of syngeneic or allogeneic mesenchymal stem cells as cell carriers and the specific role of the immune system in this therapy are still unknown. In this study we use our virotherapy Celyvir with syngeneic and allogeneic mouse mesenchymal stem cells to determine their antitumor efficacy in a C57BL/6 murine adenocarcinoma model. Adoptive transfer of splenocytes from treated mice to new tumor-bearing mice followed by a secondary adoptive transfer to a third group was performed. Similar reduction of tumor growth and systemic activation of the innate and adaptive immune system was observed in groups treated with syngeneic or allogeneic mesenchymal stem cells loaded with ICOVIR-5. Moreover, a different pattern of infiltration was observed by immunofluorescence in Celyvir-treated groups. While non-treated tumors presented higher density of infiltrating immune cells in the periphery of the tumor, both syngeneic and allogeneic Celyvir-treated groups presented higher infiltration of CD45+ cells in the core of the tumor. Therefore, these results suggest that syngeneic and allogeneic Celyvir induce systemic activation of the immune system, similar antitumor effect and a higher intratumoral infiltration of leukocytes.
KeywordsOncolytic virus Mesenchymal stem cells Tumor infiltration Immune response Celyvir Immunotherapy
Mesenchymal stem cells infected with ICOVIR-5
Human mesenchymal stem cells
Murine mesenchymal stem cells infected with ICOVIR-5
Murine mesenchymal stem cells
Multiplicity of infection
Mesenchymal stem cells
Phosphorylated Akt protein
The authors would like to thank Isabel Cubillo, Alicia Giménez and Elena Calvo for their technical support in the study.
Álvaro Morales-Molina conducted and analyzed the experiments shown in this paper, contributed to the experimental design and participated in the writing of the paper; Stefano Gambera contributed to the tumor growth studies shown in this paper; Teresa Cejalvo participated in the immune analysis by flow cytometry; Rafael Moreno participated in the writing and revision of the paper; Miguel Ángel Rodríguez-Milla performed the western blot assays shown in this paper; Ana Judith Perisé-Barrios contributed to the IHC assays shown in this paper and the experimental design; and Javier García-Castro participated in the overall project and experimental design, and participated in the writing of the paper.
This study was funded by Ministerio de Economía y Competitividad of Spain (PI14CIII/00005 and PI17CIII/00013 grants to Javier García-Castro); Consejería de Educación, Juventud y Deporte of Comunidad de Madrid (P2010/BMD-2420 grant); Fundación Oncohematología Infantil (CIF G83770297), AFANION (CIF G02223733), and Asociación Pablo Ugarte (CIF G86121019), whose support we gratefully acknowledge.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
Ethical approval and ethical standards
All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving animals were approved by the Animal Research and Welfare Ethics Committee (Comité de Ética de la Investigación y de Bienestar Animal) of Instituto de Salud Carlos III (Ref: PROEX 347/15), where the studies were conducted. This article does not contain any studies with human participants.
Research involving animals source
C57BL/6 and C57BL/10 mice were bred in the animal facility of the Instituto de Salud Carlos III, Majadahonda, Spain. All animals were housed under specific pathogen-free conditions in accordance with the guidelines detailed in Royal Decree 52/2013 of Spain.
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