The anticancer effect of potential probiotic L. fermentum and L. plantarum in combination with 5-fluorouracil on colorectal cancer cells.

5-Fluorouracil (5-FU) is an effective chemotherapy drug in the treatment of colorectal cancer (CRC). However, auxiliary or alternative therapies must be sought due to its resistance and potential side effects. Certain probiotic metabolites exhibit anticancer properties. In this study evaluated the anticancer and potential therapeutic activities of cell extracts potential probiotic strains, Limosilactobacillus fermentum and Lactiplantibacillus plantarum isolated from the mule milk and the standard probiotic strain Lacticaseibacillus rhamnosus GG (LGG) against the human colon cancer cell line (HT-29) and the normal cell line (HEK-293) alone or in combination with 5-FU. In this study, L. plantarum and L. fermentum, which were isolated from mule milk, were identified using biochemical and molecular methods. Their probiotic properties were investigated in vitro and compared with the standard probiotic strain of the species L. rhamnosus GG. The MTT assay, acridine orange/ethidium bromide (AO/EB) fluorescent staining, and flow cytometry were employed to measure the viability of cell lines, cell apoptosis, and production rates of Th17 cytokines, respectively. The results demonstrated that the combination of lactobacilli cell extracts and 5-FU decreased cell viability and induced apoptosis in HT-29 cells. Furthermore, this combination protected HEK-293 cells from the cytotoxic effects of 5-FU, enhancing their viability and reducing apoptosis. Moreover, the combination treatment led to an increase in the levels of IL-17A, IFN-γ, and TNF-α, which can enhance anti-tumor immunity. In conclusion, the cell extracts of the lactobacilli strains probably can act as a potential complementary anticancer therapy.

Formalin and ferric chloride inactivated Pasteurella multocida type a adjuvanted with bacterial DNA and alum as a new vaccine candidate in sheep pasteurellosis.

The aim of the present study was to investigate humoral and cellular immune responses in sheep inoculated with inactivated P. multocida antigen with alum and bacterial DNA adjuvant by identifying IgG and cytokines from serum and cell culture. Sheep were immunized with iron and formalin-inactivated antigens at an interval of 2 weeks. These immunogens were mixed with alum adjuvant and P. multocida type A DNA (AbDNA). After injection and blood sampling, the serum antibody titer and cellular immune responses (IL-4, IFN-γ, and TNF-α) on serum samples and lymphocyte cell were tested by ELISA. The ELISA results showed a higher antibody titer in the bDNA adjuvant group compared to the alum adjuvant group and the control group. In general, the level of IgG in the serum of immunized animals was significantly increased compared to the control group. The peak antibody titer (1.794) was observed on the 28th day of injection in the IIV-AbDNA group. After immunization, inactivation with iron and bDNA adjuvant increased cytokine production compared to other experimental and control groups. High levels of lymphocyte and serum titers of IL-4, IFN-γ, and TNF-α were also obtained in the IIV-AbDNA group. The findings showed that killed P. multocida type A antigens formulated with bacterial DNA as an adjuvant are candidates for new immunogens against P. multocida infections in sheep. The inactivation of bacteria with iron also enhanced proper immune responses.