Effect of capsular polysaccharide and detoxified lipopolysaccharide Shigella sonnei on tumor node growth and cytokine synthesis in C57Bl/6 mice after inoculation of melanoma B16

Abstract

Introduction. Melanoma represents an immunogenic tumor which cells are recognized by the immune system. Under certain conditions the immune system can suppress the development of malignant neoplasm, as evidenced by clinical observations of spontaneous regression and complete elimination of the tumor in some patients.

One of the possible approaches to the induction of regression of malignant tumors is the stimulating the cells of the immune system with products of bacterial origin. In this stimulation Toll-like receptors play a key role, recognizing the conservative structures of microorganisms and activating the innate and adaptive immune response.

Aim of the study – to investigate the ability of the two bacterial compounds: detoxified lipopolysaccharide (Ас3-S-LPS) and capsular polysaccharide (CPS) of Shigella sonnei phase 1 to stimulate antitumor immune response in experimental animals with transplanted malignant melanoma.

Material and methods. The transplanted melanoma cell line B16 was used as a model for the study. The B16 melanoma cell line was provided by the Cellular Immunity Laboratory, Research Institute of Experimental Diagnostics and Tumor Therapy (NII EDiTO), N.N. Blokhin NMRCO, MOH of Russia. The antitumor activity of the drugs was estimated by the effect on the growth of primary tumor nodes in mice C57Bl/6J after introduction of tumor cells. When studying the mechanisms of antitumor activity, the effect of drugs on the formation of nitric oxide and the synthesis of cytokines (IL-1β, IL-6, IL-10, TNF and GM-CSF) was assessed.

Results. It has been shown that the administration of Ac3-S-LPS and CPS to mice leads to a slowdown in the growth of primary tumor nodes (PTN), which is not associated with a direct cytotoxic effect on tumor cells. Ac3-S-LPS and CPS, as well as Shigella sonnei native LPS, are unable to induce nitric oxide synthesis by B16 melanoma cells. However Ac3-S-LPS and native LPS induce nitric oxide synthesis in PTN cell culture. We have shown that B16 melanoma cells when cultured in vitro spontaneously produce three of five studied cytokines (IL-1β, IL-6 and IL-10) and do not synthesize TNF and GM-CSF. The addition of Shigella sonnei LPS or Ac3-S-LPS to cells stimulated the synthesis of IL-1β, IL-6, IL-10 and did not affect the synthesis of the other two cytokines.

The cytokines TNF, GM-CSF, and IL-4 were detected in the supernatants of PTN cultures. Their concentration increased when Ac3-S-LPS was added to cultures. The introduction of the Ac3-S-LPS preparation directly into the PTN led to the stimulation of IL-10 synthesis and did not affect the synthesis of other cytokines.

Conclusion. The antitumor activity of 2 bacterial compounds was studied on the model of transplanted melanoma: Ac3-S-LPS and CPS of Shigella sonnei phase 1 microorganisms, which have not been previously studied in this aspect. The obtained results allow us to classify them as TLRs agonists that suppress the growth of B16 malignant melanoma tumor after inoculation in experimental animals as a result of stimulation of innate immune responses. The study of the effect of cytokines induced by the interaction of bacterial ligands with TLRs on immune system and tumor cells leads to a deeper understanding of the pathophysiology of carcinogenesis and may contribute to the development of targeted therapy techniques for inhibiting tumor growth.

Keywords:antitumor activity; B16 melanoma; ligands; Toll-like receptor; lipopolysaccharide; cytokines

For citation: Novikova E.M., Kozyreva O.V., Razvalyayeva N.A., Chukhina E.S., Golovina M.E., L’vov V.L., Aparin P.G., Stepanenko R.N. Antitumor activity of ligands of Toll-like receptors of bacteria Shigella sonnei. Immunologiya. 2023; 44 (2): 167–80. DOI: https://doi.org/10.33029/0206-4952-2023-44-2-167-180 (in Russian)

Funding. The study had no sponsor support.

Conflict of interests. The authors declare no conflict of interests.

Authors’ contribution. Study conception and design – Stepanenko R.N.; material collection and processing – Novikova E.M., Kozyreva O.V., Razvalyaeva N.A., Chukhina E.S., Golovina M.E., L’vov V.L., Aparin P.G., Stepanenko R.N.; statistical processing – Kozyreva O.V., Chukhina E.S., Stepanenko R.N.; article writing – Novikova E.M., Kozyreva O.V., Stepanenko R.N.; article editing – Chukhina E.S., L’vov V.L., Aparin P.G., Stepanenko R.N.

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