Autologous dendritic cells and activated cytotoxic T‐cells as combination therapy for breast cancer

Cytotoxicity against MCF-7

Cytotoxicity against MCF-7



Breast cancer is the most common oncological pathology in women worldwide. A type of treatment applying dendritic cells (DCs) and cytotoxic DC‑induced antigen‑specific T lymphocytes efficiently eliminates residual cancer cells that are the key cause of tumor recurrence and metastasis.The results of the present study demonstrated that the investigated cellular immunotherapy for breast cancer is safe, reduces the risk of relapse and metastasis, and improves immunity by reducing the number of regulatory T cells.


The study involved 25 patients ages 18-65 years old with histologically diagnosed with Breast cancer, with adequate bone marrow, liver and renal function, negative for human immunodeficiency virus and hepatitis B and C virus.

Preparation of tumor cell lysates.
Tumor cell lysates were prepared from tissue samples after radical surgery or incisional biopsy.


A 1-3-cm3 tumor tissue sample was placed into a sterile tube, the adjacent (macroscopically unaltered) tissues were removed, and the sample was subjected to mechanical homogenization and four-freeze thaw cycle (-80oC and room temperature, respectively). Larger particles were removed by centrifugation at 266 x g and 24oC (room temperature) for 2 minutes. The lysate was filter-sterilized (filter pore diameter = 0.45 um) and the protein concentration of the lysate was measured using a NanoDrop 2000. The tumor cell lysate was divided into aliquots, frozen and stored -80oC.


DC preparation and characterization:

Mononuclear cells were isolated from the peripheral blood of the patients using Ficoll gradient centrifugation. The isolated peripheral blood mono- nuclear cells (PBMCs) were washed in RPMI- 1640 medium and centrifuged twice at 266 x g at 24oC (room temperature) for 10 min. Cells were cultured in RPMI-1640 medium supplemented with 10% FBS, 2 mM L-glutamine, 10 mM HEPES, 80 ug/ml gentamycin, 100 ug/ ampicillin. Mononuclear cells (1-1.5×106 cells/ml) in RPMI-1640 complete medium supplemented with 10% FBS were placed into 150-cm2 (690 mL) culture flasks with vintage caps. The cells were allowed to adhere to the flask in CO2 incubator at 37OC and 100% humidity for 30 minutes. Viable non-adherent mononuclear cells (non-adherent PBMCs) were cultured in complete RPMI-1640 medium with partial media changes on days 3 and 5. The adherent cells were removed using a cell scraper and washed with RPMI-1640 medium.

The adherent cell fraction was used to generate mature antigen activated DCs. For this purpose, 100 ng/ml granulocyte-macrophage colony-stimulating factor (GM-CSF) and 50 ng/ml IL-4 were added to the cells, which were cultured in a 75-cm2 (270mL) culture flask in complete RPMI-1640 medium supplemented with 10% FBS for 72 h to generate immature DCs. The tumor tissue lysate was added to immature DC for 24
hours. To obtain mature DCs, TNF-a was added to the fresh medium within 48 hours.


Clinical activity for patients with stage IV or progressive disease:

Among patients with stage IV disease, 2 patients remained stable for 6 months. 1 patient developed local progresion, distant foci were undetectable, and chemotherapy was not administered. 1 patient succumbed to disease progression (two injections were administered). Three lines of chemotherapy did not achieve positive response.

Side effects:
The treatment was generally well-tolerated. The patients were monitored by medical personnel for 24 h after injection to assess toxicity. The most common adverse event were flu-like symptoms, such as fever and fatigue, that did not require additional treatment or prolonged hospitalization. All symptoms spontaneously resolved without treatment after 2-3 h.


This result demonstrated that the investigated cellular immunotherapy for breast cancer is safe, reduces the risk of relapse and metastasis, and improves immunity by reducing the number of regulatory T cells. Therefore, this therapeutic strategy may represent a novel approach to combating distant metastases of breast cancer. Dendritic cell and Activated T cells in combination with other therapies is effective and prolonged life of patients with breast cancer.

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Scientific article publishing date : 22/11/2019

Immucura identifier : BSC21_026EN