Copyright: © 2024 by the authors. Licensee: Pirogov University.
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ORIGINAL RESEARCH

Interferon type I-expressing recombinant vaccinia virus as a platform for selective immunotherapy of glioblastoma and melanoma

About authors

1 Engelhardt Institute of Molecular Biology, Russian Academy of Science, Moscow, Russia

2 Federal Scientific and Clinical Center of Specialized Types of Medical Care and Medical Technologies, Federal Medical Biological Agency of Russia, Moscow, Russia

3 Federal Center of Brain Research and Neurotechnologies of the Federal Medical Biological Agency, Moscow, Russia

4 Research Institute of Pulmonology of the Federal Medical Biological Agency, Moscow, Russia

Correspondence should be addressed: Anastasia V. Lipatova
Vavilova, 32/1, Moscow, 119991, Russia; moc.liamg@vnaavotapil

About paper

Funding: the design of recombinant vaccinia virus strains was supported by the Russian Science Foundation grant No. 23-14-00370, and assessment of their properties in in vitro and in vivo models was supported by the Russian Science Foundation grant No. 22-64-00057; histologic and immunohistochemical assessment of tumor tissue was supported by FMBA of Russia.

Author contribution: Naberezhnaya ER — implementation of in vitro and in vivo experiments, manuscript writing; Soboleva AV — flow cytometry data acquisition, determining the cell line sensitivity to viruses; Vorobyev PO — designing recombinant viruses, production of preparative amounts of strains; Vadekhina VV — conducting in vivo experiments; Yusubalieva GM — interpretation of in vivo experimental data, manuscript writing; Isaeva IV — histologic and immunohistochemical assessment; Baklaushev VP — microscopy, describing histology and immunohistochemistry data, preparing a drawing, manuscript writing; Chumakov PM — manuiscript editing; Lipatova AV — study concept, general project management.

Compliance with ethical standards: the in vivo study was approved by the Ethics Committee of the Federal Scientific and Clinical Center of FMBA of Russia (protocol No. 7 dated 06 September 2022) and conducted in accordance with the the Eurasian Economic Commission Board’s guidelines No. 33 dated 14 November 2023 "On the Guidelines for handling laboratory (experimental) animals when conducting preclinical (non-clinical) studies". The number of animals per group was minimized; the subcutaneous tumor size in the groups did not exceed 2000 mm3. In vitro experiments involved the commercially available animal and human cell lines.

Received: 2024-11-26 Accepted: 2024-12-19 Published online: 2024-12-30
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Fig. 1. Schematic illustration of the plasmid design
Fig. 2. Photographs of BHK-21 cells 24 h after infection with LIVP-mIFNα (A–C) and LIVP-hIFNα (D–F) (A and D — light field; B and E — red fluorescent channel; C and F — combined image). 200x magnification; scale bar = 100 μm
Fig. 3. Evaluation of virus replication kinetics 24, 48, 72 and 96 h after infection with recombinant VV strain on different cell lines
Fig. 4. Sensitivity of tumor and normal cell lines to vaccinia virus expressing tagRFP and a) human IFNα and b) mouse IFNα. X-axis: tumor cell lines, Y-axis: lgTCID50/ml
Fig. 5. Evaluation of functional activity of interferons expressed by LIVP-hIFNα strains on U87-MG cell line (A) and LIVP-mIFNα on the B16 cell line (B)
Fig. 6. Dynamics of tumor growth after LIVP-mIFNα injection
Fig. 7. Histological and immunohistochemical examination of tumor preparations 24 days after inoculation. A. Hematoxylin and eosin staining (general view of the tumor on the left side images and fragments at ×200 magnification on the right side images). B. Immunohistochemical staining for differentiation clusters: CD4, CD8, CD56. Representative preparations of animals from the control group, the group with LIVP-RFP, and the LIVP-mIFNα group, respectively, are shown from top to bottom.