The development of new research techniques, especially molecular ones, creates hopes for improved treatment efficacy and a better prognosis in lung cancer. The starting point for very important experiments in the field of immunotherapy is the sensitization of the immune system to the tumor antigen.

The aim of the study was to determine whether using flagellin can induce innate antitumor effective, and thus, increase the immunogenicity of the tumor.

The test material was the established mouse cell line LLC, derived from Lewis lung cancer. Vectors were constructed by cloning FliC inserts into the pCDH-MSC-T2A Puro plasmid. The cells were transfected directly and indirectly (with pseudoviruses, produced by previously transfected packaging cells HEK 293T). The transfection efficiency was confirmed by RT-PCR. The cells thus prepared were implanted subcutaneously in mice. The control group received non-transfected LLC cells. Then, after 6 weeks, the mice were scarified. The animals were sectioned to isolate the tumor and lungs in which the presence or absence of metastases was assessed.

In mice that received immunotherapeutic vaccines containing transfected LLC cells, less tumor mass was found or a complete lack of it, and the lifespan was noticeably prolonged. In addition, no metastases occurred in the group vaccinated with cells transfected with direct technique.

It can be concluded that flagellin is effective as an adjuvant in the process of presenting tumor antigens to T cells Thus, in the light of recent studies and reports, it is likely that in this experiment effectively increased immunogenicity of the tumor. Activating anti-tumor cellular responses using flagellin is a promising target for lung cancer immunotherapy.

Tokarz−Deptuła B, Niedźwiedzka P, Deptuła W.: Nowe receptory w immunologii. Centaur Lubuski, 2005; 63: 12–15

Akira, S, Hemmi H.: Recognition of pathogen-associated molecular patterns by TLR family. Immunology Letters, 2003; 85: 85-95

Majewska M., Szczepanik M.: Rola receptorów toll-podobnych (TLR) w odporności wrodzonej i nabytej oraz ich funkcja w regulacji odpowiedzi immunologicznej. Postępy Higieny i Medycyny Doświadczalnej, 2006; 60: 52-63

Takeuchi O, Akira S.: Genetic approaches to the study of Toll-like receptor function. Microbes and Infection, 2002; 4: 887-895

Banchereau J., Palucka A.K.: Dendritic cells as therapeutic vaccines against cancer. Nature Reviews Immunology, 2005; 5: 296–306

Saloga J, Enk AH, Ross R.: Dendritic cells in the allergic immune response- from a key player in the pathological process to a potential therapeutic tool. ACI International, 2001; 13: 107-112

Seya T, Akazawa T, Tsujita T.: Role of Toll-like Receptors in Adjuvant-Augumented Immune Therapies. 2006; Evidence-based Complementary and Alternative Medicine 3(1): 31-38

Didierlaurent Arnaud., Ferrero I., Otten L.A.: Flagellin Promotes Myeloid Differentiation Factor 88-Dependent Development of Th2-Type Response. The Journal of Immunology 2004; 172: 6922–6930

Ivison S.M., Khan M.A.S., Graham N.R.: A phosphorylation site in the Toll-like receptor 5 TIR domain is required for inflammatory signaling in response of flagellin. Biochemical and Biophysical Research Communications, 2007; 352: 936-941

Steiner TS: How flagellin and toll-like receptor 5 contribute to enteric infection. Infection and Immunity, 2007; 75(2): 545-52

Applequist S.E., Rollman E., Wareing M.D.: Activation of Innate Immunity, Inflammation, and Potentiation of DNA Vaccination through Mammalian Expression of the TLR5 Agonist Flagellin. Journal of Immunology, 2005; 175: 3882-3891

Akira S., Takeda K.: Toll-like receptor signalling. Nature Reviews Immunology, 2004; 4: 499-511

Sfondrini L, Rossini A, Besusso D.: Antitumor Activity of the TLR-5 Ligand Flagellin in Mouse Models of Cancer. Journal of Immunology, 2006; 176: 6624-6630

Tsujimoto H, Uchida T, Efron PA: Flagellin engances NK cell proliferation and activation directly and though dendritic cell-NK cell interactions. Journal of Leukocyte Biology, 2005; 78: 888-897

Means T. K, Hayashi F, Smith K. D.: The Toll-Like Receptor 5 Stimulus Bacterial Flagellin Induces Maturation and Chemokine Production in Human Dendritic Cells. Journal of Immunology, 2003; 140: 5165-5175

http://www3.bio-rad.com/B2B/BioRad/product

USPTO Patent Application 20080248068, Use of flagellin as an adjuvant for vaccine

Agrawal S., Agarawal A., Doughty B.: Different Toll-Like Receptor Agonists Instruct Dendritic Cells to Induce Distinct Th Responses via Differential Modulation of Extracellular Signal-Regulated Kinase-Mitogen-Activated Protein Kinase and c-Fos. Journal of Immunology, 2003; 171: 4984-4989

McSorley S.J., Ehst B.D., Yu Y.: Bacterial Flagellin Is an Effective Adjuvant for CD4+ T Cells In Vivo. Journal of Immunology, 2002; 169: 3914–3919

Salazar-Gonzalez RM, Srinivasan A, Griffin A.: Salmonella Flagellin Induces Bystander Activation of Splenic Dendritic Cells and Hinders Bacterial Replication In Vivo. Journal of Immunology, 2007; 179: 6169-6175

Cuadros C., Lopez-Hernandez F., Dominguez A.L.: Flagellin Fusion Proteins as Adjuvants or Vaccines Induce Specific Immune Responses. Infection and Immunity 2004

Vicente-Suarez I., Brayer J., Villarga A.: TLR5 ligation by flagellin converts tolerogenic dendritic cells into activating antigen-presenting cells that preferentionally induce T-helper 1 responses. Immunology Letters, 2009; 125(2):114-118