A Comprehensive Analysis of Cytotoxicity of Genetically Modified Lactobacillus plantarum 8РАЗ (B-11007) Strain Including Identification and Evaluation of Virulence Factors

Introduction. Active development of biotechnologies and genetic engineering techniques broadens the horizons for engineering the probiotic bacterial strains with tailored functional properties, including the strains based on bacteria of the genus Lactobacillus that are traditionally considered safe microorganisms. Genetic modification enhances their antagonist activity, resistance to adverse environmental factors, and boosts metabolic efficiency. However, bacterial genome editing may entail risk of introducing the undesirable traits, including cytotoxicity and expression of virulence factors. Safety concerns are particularly relevant in the frame of veterinary practice, where probiotics are widely used to correct the gastrointestinal microbiocenosis in agricultural animals and improve their productivity. The aim of the study is to conduct a comprehensive analysis of cytotoxicity of the genetically modified Lactobacillus plantarum 8PAЗ (B-11007) strain including identification and evaluation of the virulence factors responsible for inducing cell death and disruption of cellular functions.
Materials and Methods. A genetically modified Lactobacillus plantarum 8PAЗ (B-11007) strain was the object of the study conducted in the period from January to February 2026. A comprehensive in vitro analysis of its cytotoxicity included: assessment of the cytopathic effect in intestinal cell culture, the effect on the viability and metabolic activity of enterocytes, and the state of intestinal epithelial barrier function. Additionally, the analysis of potential virulence factors was performed, along with the study of the mechanisms of cell death, including signs of apoptosis and necrosis.
Results. The recombinant Lactobacillus plantarum 8PAЗ (B-11007) strain has demonstrated a high biological safety and pronounced biological activity, which enhanced the intestinal epithelial barrier function by 15.2%, the metabolic activity of cells by 8.7% and simultaneously reduced early apoptosis of enterocytes by 31.1% (p<0.05), showing no signs of cytotoxic action or activation of caspase-dependent cell death.
Discussion and Conclusion. The conducted comprehensive analysis allows making the unequivocal conclusion about genetically modified L. plantarum 8PAЗ (B-11007) strain exhibiting no cytotoxic activity in vitro. Moreover, by certain properties (cellular metabolic activity, barrier function), it exceeds the original native strain, which makes it a promising candidate for further preclinical and clinical trials as a potential probiotic agent.

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