Purpose: Breast cancer, a leading cause of death in female cancer types, often poses a high metastasis risk. The study focuses on enhancing antitumor immunotherapy using dendritic cells (DCs), vital for initiating and coordinating adaptive immune responses. Additionally, gamma delta (γδ) T lymphocytes, known for their cytokine secretion (Interleukin 17 [IL-17], Interferon gamma [IFN-γ]) and cytotoxicity against various cells, are examined. Methods: To evaluate the effectiveness of different DC maturation protocols, female BALB/c mice were divided into three groups: Tumor (n=12), Protocol 1 (n=12, maturation with TNF-α), and Protocol 2 (n=12, maturation with TNF-α, IL-12, and RANTES). All were induced to develop tumors using the 4T1 cell line, followed by treatment with either Protocol 1 or 2. The quantification of cytotoxic T lymphocytes (CD8+) producing IFN-γ and γδ T lymphocytes producing IL-17 and IFN-γ was conducted using flow cytometry. Results: Findings revealed a higher number of cytotoxic T lymphocytes producing IFN-γ and γδ T lymphocytes producing IL-17 and IFN-γ in mice treated with Protocol 2 compared to Protocol 1. This suggests Protocol 2’s superior efficacy in DC maturation for antitumor treatment. Conclusion: Protocol 2 demonstrates greater efficiency in DC maturation for antitumor purposes. However, being an experimental study, further research is necessary to fully understand the optimal dendritic cell maturation protocol and the role of IL-17 and IFN-γ cytokines in combating tumor cells.

Assessing the Natural Killer (NK) cells in Dendritic Cell (DC) vaccine immunotherapy of 4T1 cell-induced breast cancer in Balb/C mice, we watch the expression of killer activation receptors NKp46, NKG2D, and inhibitory killer cell immunoglobulin-like receptor (KIR) Ly49G2, NKG2A/C/E and the interleukins (IL) 2, 10, 12, 17, tumor necrosis factor alpha (TNF-α) and interferon gamma (IFN-γ). 30 Balb/C mice divided into untreated, or control experimental group (GI), untreated induced tumor group (GII), and the DC treated vaccine-induced tumor group (GIII). Bone marrow from donor mice was collected to produce the vaccine. The 25-day experiment comprised tumor volume measurements and at the end of the treatment protocol mice were euthanized for removal of the spleen and evaluation of tumor infiltration by flow cytometry of the receptors NKp46, NKG2D, Ly49G2 and NKG2A/C/E and the cytokines. Our results showed that mice treated with the DC vaccine reduced the expression of inhibition receptors Ly49G2D/NKG2A/C/E in the spleen and tumor infiltrate (p=0.0001), with increased fluorescence intensity of splenic IL-10, IL-2, IL-12, IL-17, TNF-α and IFN-γ-producing NKp46/NKG2D cells (p=0.0001), intratumoral TNF-α and IFN-γ-producing by NKp46/NKG2D cells (p=0.0001). We demonstrated that immunotherapy with DC can alter the profile of NK cells by directly influencing them into an antitumor response due to reduced expression of inhibition receptors and increased production of cytokines such as IL-2, IL-12, TNF-α and IFN-γ by cells. Also, the activation of receptors confers NK cells the ability to reverse tumor-induced immunosuppression, stimulating the immune system to eliminate tumor cells.