RESEARCH PAPER
Investigation of materials for the development of new molecular and cellular antiviral and antimalignancy vaccines and immunization methods: a pilot study
 
More details
Hide details
1
Institute of Experimental Morphology, Pathology and Anthropology with Museum to Bulgarian Academy of Sciences, Sofia, Bulgaria
 
2
Department of Pathophysiology, Medical University of Sofia, Bulgaria
 
3
Department of Toxicology, University Hospital “Joanna Queen”, Sofia, Bulgaria
 
4
Department of Biology, Medical University of Sofia, Bulgaria
 
5
Department of Pharmacology and Toxicology, Medical University of Sofia, Bulgaria
 
 
Submission date: 2025-07-02
 
 
Final revision date: 2025-09-03
 
 
Acceptance date: 2025-09-05
 
 
Corresponding author
Iskra Sainova   

Bulgarian Academy of Sciences (BAS), Bl. 61, “Hemus” St., 1574, Sofia, Bulgaria
 
 
 
KEYWORDS
TOPICS
ABSTRACT
Background:
Changes in biomolecules under the influence of chemical and physical factors on cells, tissues, and whole organisms are investigated.

Material and methods:
In vitro-incubated bovine embryonic cells were inoculated with low titers (high dilutions of viral suspensions) of vaccine avipoxviral strains. Mouse embryonic fibroblasts were co-cultivated with mouse malignant myeloma cells (P3-X63-Ag8) transfected by recombinant DNA plasmid or preincubated in culture fluid from prior incubation of the same cells. Sub-populations of virus-inoculated, co-cultivated, and preincubated cell cultures were frozen in the presence of the cryoprotectant dimethylsulfoxide (DMSO), subsequently thawed, and re-incubated. Newly formed cell monolayers were inoculated with extracellular and intracellular forms of each viral strain, both before and after exposure to DMSO and drastic temperature changes. Extracellular forms were derived from the cultural fluids of inoculated cell cultures, while intracellular forms were obtained from suspensions of mechanically scraped virus-inoculated cells.

Results:
Exchange of nucleotide (DNA and/or RNA) fragments between cellular and viral genomes, as well as between genomes of separate cells, was suggested. These changes were explained by activated fusion induced by the organic detergent (DMSO) combined with drastic temperature changes. Such processes could provide vectors for gene-engineering manipulations and the development of molecular (DNA-based, RNA-based, and/or protein-based) antiviral and antimalignant vaccines. Production of immune molecules by nonimmune cell types under appropriate conditions, such as the presence of immunomodulators, was also proposed.

Conclusions:
The results suggest the possibility of nucleotide (DNA and/or RNA) fragment exchange between separate cells, as well as between cells and virions. Nonimmune cells demonstrated the capacity to produce immune molecules under appropriate conditions.
REFERENCES (45)
1.
Acevedo OA, Berrios RV, Rodríguez-Guilarte L, Lillo-Dapre­mont B, Kalergis AM. 2021. Molecular and cellular mechanisms modulating trained immunity by various cell types in response to pathogen encounter. Front Immunol. 12: 745332. https://doi.org/10.3389/fimmu.....
 
2.
Ahmad I, Valverde A, Ahmad F, Naqvi AR. 2020. Long noncoding RNA in myeloid and lymphoid cell differentiation, polarization and function. Cells 9(2): 269. https://doi.org/10.3390/cells9....
 
3.
Barrette S, Douglas JL, Seidel NE, Bodine DM. 2000. Lentivirus-based vectors transduce mouse hematopoietic stem cells with similar efficiency to Moloney murine leukemia virus-based vectors. Blood 96(10): 3385–3391.
 
4.
Brachmann C, Davies A, Cost GJ, Caputo E, Li J, Hieter P, Boeke JD. 1998. Designer deletion strains derived from Saccharomyces cerevisiae S288C: a useful set of strains and plasmids for PCR-mediated gene disruption and other applications. Yeast 14(2): 115–132.
 
5.
Breschi A, Gingeras T, Guigó R. 2017. Comparative trans­criptomics in human and mouse. Nat Rev Genet. 18(7): 425–840.
 
6.
Chen S, Agarwal A, Glushakova OY, Jorgensen MS, Salgar SK, Poirier A, Flotte TA, Croker BP, Madsen KM, Atkinson MA, et al. 2003. Gene delivery in renal tubular epithelial cells using recombinant adeno-associated viral vectors. J Am Soc Nephrol. 14(4): 947–958.
 
7.
Cheng L, Du C, Lavau C, Chen S, Tong J, Chen BP, Scollay R, Hawley RG, Hill B. 1998. Sustained gene expression in retrovirally transduced, engrafting human hematopoietic stem cells and their lympho-myeloid progeny. Blood 92(1): 83–92.
 
8.
Domi A, Moss B. 2005. Engineering of a vaccinia virus bacterial artificial chromosome in Escherichia coli by bacteriophage-based recombination. Nat Methods. 2(2): 95–97.
 
9.
Frey S, Marsh M, Günther S, Pelchen-Matthews A, Stephens P, Ortlepp S, Stegmann T. 1995. Temperature dependence of cell–cell fusion induced by the envelope glycoprotein of human immunodeficiency virus type 1. J Virol. 69(3): 1462–1472.
 
10.
Galindo I, Lorenso M, Blasco R. 2001. Set of vectors for the expre­ssion of histidine-tagged proteins in vaccinia virus recombinants. Biotechniques. 30(3): 524–526.
 
11.
Goldstein ME, Scull MA. 2022. Modeling innate antiviral immunity in physiological context. J Mol Biol. 434(6): 167374. https://doi.org/10.1016/j.jmb.....
 
12.
Gonçalves MAFV. 2005. Adeno-associated virus: from defective virus to effective vector. Virol J. 2: 43. https://doi.org/10.1186/1743-4....
 
13.
Guenechea G, Gan OI, Inamitsu T, Dorell C, Pereira DS, Kelly M, Naldini L, Dick JE. 2000. Transduction of human CD34+ CD38– bone marrow and cord blood-derived SCID-repopulating cells with third-generation lentiviral vectors. Mol Ther. 1(6): 566–573.
 
14.
Hamada A, Torre C, Drancourt M, Ghogo E. 2019. Trained immunity carried by non-immune cells. Front Microbiol. 9: 3225. https://doi.org/10.3389/fmicb.....
 
15.
Jacob SW, Herschler R. 1986. Pharmacology of DMSO. Cryobiology. 23(1): 14–27.
 
16.
Johnson PA, Wu A, Johnson JC, Schauer Z, Wu T, Fernandes F, Schabert R, Mardon A. 2022. Advances in DNA- and RNA-based oncolytic viral therapeutics and immunotherapies. Appl Microbiol. 2(2): 319–329.
 
17.
Kang H, Ga YJ, Kim SH, Cho YH, Kim JW, Kim C, Yeh JY. 2023. Small interfering RNA (siRNA)-based therapeutic applications against viruses: principles, potential, and challen­ges. J Biomed Sci. 30(1): 88. https://doi.org/10.1186/s12929....
 
18.
Kao CY, Mills JA, Burke CJ, Morse B, Marques BF. 2023. Role of cytokines and growth factors in the manufacturing of iPSC-derived allogenic cell therapy products. Biology 12(5): 677. https://doi.org/10.3390/biolog....
 
19.
Kearney JF, Radbruch A, Liesegang B, Rajewsky K. 1979. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 123(4): 1548–1550.
 
20.
Kobari L, Pflumio F, Giarratana M, Li X, Titeux M, Izac B, Leteurtre F, Coulombel L, Douay L. 2000. In vitro and in vivo evidence for the long-term multilineage (myeloid, B, NK, and T) reconstitution capacity of ex vivo-expanded human CD34+ cord blood cells. Exp Hematol. 28(12): 1470–1480.
 
21.
Kyba M, Perlingeiro RC, Hoover RR, Lu C-W, Pierce J, Daley GQ. 2003. Enhanced hematopoietic differentiation of embryo­nic stem cells conditionally expressing Stat5. Proc Natl Acad Sci USA. 100(1): 11904–11910.
 
22.
Li G, Peñagaricano F, Weigel KA, Zhang Y, Rosa G, Khatib H. 2012. Comparative genomics between fly, mouse, and cattle identifies genes associated with sire conception rate. J Dairy Sci. 95(10): 6122–6129.
 
23.
Li Z, Zhang X, Liu C, Ma J. 2021. Non-immune cell components in the gastrointestinal tumor microenvironment influencing tumor immunotherapy. Front Cell Dev Biol. 9: 729941. https://doi.org/10.3389/fcell.....
 
24.
Lundstrom K. 2019. RNA viruses as tools in gene therapy and vaccine development. Genes (Basel) 10(3): 189. https://doi.org/10.3390/genes1....
 
25.
Maguire G. 2021. Stem cells part of the innate and adaptive immune systems as a therapeutic for COVID-19. Commun Integr Biol. 14(1): 186–198.
 
26.
Marshall JS, Warrington R, Watson W, Kim HL. 2018. An intro­duction to immunology and immunopathology. Allergy Asthma Clin Immunol. 14(2): 49. https://doi.org/10.1186/s13223....
 
27.
McLaughlin SK, Collis P, Hermonat PL, Muzyczka N. 1988. Adeno-associated virus general transduction vectors: ana­lysis of proviral structures. J Virol. 62(6): 1963–1973.
 
28.
de Ménorvan MA, Mir LM, Fernández LM, Reigada R. 2012. Effects of dimethyl sulfoxide in cholesterol-containing lipid membranes: a comparative study of experiments in silico and with cells. PLoS One. 7(7): e41733. https://doi.org/10.1371/journa....
 
29.
Murray MJ, Peters NE, Reeves MB. 2018. Navigating the host cell response during entry into sites of latent cytomegalovirus infection. Pathogens 7(1): 30. https://doi.org/10.3390/pathog....
 
30.
Neeli I, Dwivedi N, Khan S, Radic M. 2009. Regulation of extra­cellular chromatin release from neutrophils. J Innate Immun. 1(3): 194–201.
 
31.
Neubert E, Meyer D, Rocca F, Günay G, Kwaczala-Tessmann A, Grandke J, Senger-Sander S, Geisler C, Egner A, Schön MP, et al. 2018. Chromatin swelling drives neutrophil extracellular trap release. Nat Commun. 9(1): 3767. https://doi.org/10.1038/s41467....
 
32.
Norwood TH, Ceigler CJ, Martin GM. 1976. Dimethyl sul­foxide enhances polyethylene glycol-mediated somatic cell fusion. Somat Cell Genet. 2(3): 263–270.
 
33.
Ochando J, Mulder WJM, Madsen JC, Netea MG, Duivenvoorden R. 2023. Trained immunity – basic concepts and contributions to immunopathology. Nat Rev Nephrol. 19(1): 23–37.
 
34.
Palese P, Roizman B. 1996. Genetic engineering of viruses and of virus vectors: a preface. Proc Natl Acad Sci USA. 93(21): 11287. https://doi.org/10.1073/pnas.9....
 
35.
Sainova I, Kolyovska V, Ilieva I, Markova TZ, Dimitrova-Dikanarova D, Hadjiolova R. 2023. The development of methods for the production of new molecular vaccines and appropriate fragments to counteract unwanted genes: a pilot study. Vaccines 11(7): 1226. https://doi.org/10.3390/vaccin....
 
36.
Sainova I, Kril A, Simeonov K, Popova T, Ivanov I. 2005. Investigation of the morphology of cell clones, derived from the mammalian EBTr cell line and their susceptibility to the vaccine avian poxvirus strains FK and Dessau. J Virol Methods. 124(1–2): 37–40.
 
37.
Schott JW, Morgan M, Galla M, Schambach A. 2016. Viral and synthetic RNA vector technologies and applications. Mol Ther. 24(9): 1513–1527.
 
38.
Segura SE, Ramos-Rivera G, Suhrland M. 2018. Educational case: infectious diseases: pathogenesis, diagnosis, treatment, and prevention. Acad Pathol. 5: 1–6.
 
39.
Sisto M, Lisi S. 2023. Immune and non-immune inflammatory cells involved in autoimmune fibrosis: new discoveries. J Clin Med. 12(11): 3801. https://doi.org/10.3390/jcm-12....
 
40.
Tercan H, Riksen NP, Joosten LAB, Netea MG, Bekkering S. 2021. Trained immunity: long-term adaptation of innate immune responses. Arterioscler Thromb Vasc Biol. 41(1): 55–61.
 
41.
Wong CM, McFall ER, Burns JK, Parks RJ. 2013. The role of chromatin in adenoviral vector function. Viruses 5(6): 1500–1515.
 
42.
Xiong C, Tang DQ, Xie CQ, Zhang L, Xu KF, Thompson WE, Chou W, Gibbons GH, Chang LJ, Yang LJ, Chen YE. 2005. Genetic engineering of human embryonic stem cells with lentiviral vectors. Neural Stem Cells Compend. 14(4): 367–377.
 
43.
Yan N, Chen ZJ. 2012. Intrinsic antiviral immunity. Nat Immunol. 13(3): 214–222.
 
44.
Zhang XY, La Russa VF, Bao L, Kolls J, Schwarzenberger P, Reiser J. 2002. Lentiviral vectors for sustained transgene expression in human bone marrow-derived stromal cells. Mol Ther. 5(5 Pt 1): 555–565.
 
45.
Zhang XY, La Russa VF, Reiser J. 2004. Transduction of bone marrow-derived mesenchymal stem cells by using lentivirus vectors pseudotyped with modified RD114 envelope glycoproteins. J Virol. 78(3): 1219–1229.
 
eISSN:2353-9461
ISSN:0860-7796
Journals System - logo
Scroll to top