PHENOTYPIC CHARACTERISTIC OF THE DENDRITIC CELLS GENERATED FROM PERIPHERAL BLOOD MONOCYTES IN PATIENTS WITH TRIPLE-NEGATIVE BREAST CANCER

Lyalkin S. A., Skachkova O. V., Gorbach O. I., Khranovska N. M., Syvak L. A., Veryovkina N. O.

CLINICAL AND EXPERIMENTAL MEDICINE

Scentific article

Triple-negative breast cancer (TNBC) is considered an aggressive cancer with the occurrence of early metastases and short follow-up period. TNBC was detected in 10-15% of breast cancer cases. The developing of the new treatment approaches for patients with TNBC is very important now. Immunotherapy (IT) is one of the perspective treatments in advanced TNBC, which is based on the activation and enhancement of the specific immune response to the tumor. Recent anti-tumor IT includes specific monoclonal antibodies (for example, checkpoint inhibitors) and adaptive cellular immunotherapy, which includes antitumor vaccines. The potential interest of anti-tumor IT is presented the natural origin adjuvants, such as dendritic cells (DC) – the main antigen-presenting cells of the immune system. The aim of the study was to investigate the quantitative and phenotypic characteristics of dendritic cells (DCs) derived from mononuclear peripheral blood cells, which may be used for antitumor therapy in TNBC patients. Object and methods. IT based on DC was administered as adjuvant treatment for patients with advanced TNBC after the basic treatment. DC was generated from peripheral blood monocytes and loaded with lysate of MDAMB-231 cells. The phenotypic analysis of the autologous generated DC and populations of peripheral blood lymphocytes was performed by flow cytometry. DCs generated from TNBC patients had slightly reduced quantitative and functional properties of maturity cells compared to cells from almost healthy people. Tumor antigens derived from MDA-MB-231 cell line can lead sufficient maturity of DCs, these data were confirmed by DCs phenotypic characteristics. The DC phenotype was analyzed by flow cytometry using monoclonal antibodies to CD83, CD86, HLA-ABC antigens labeled with FITC and antibodies to CD 11c, HLA-DR antigens labeled with PE (BectonСoulter, USA). DC phenotype was assessed at the initial (1-3 introduction of DC vaccine) and final stages (4-5 injections of DC vaccine) of IT in patients with TNBC. The phenotype analysis was performed on a FACSCalibur flow cytometer (Becton Dickinson, USA) using Cell Quest-PRO software (Becton Dickinson, USA). Results. DCs generated from peripheral blood monocytes of TNBC patients had the slightly reduced quantitative and functional parameters of maturity compared to cells in healthy people. So, the amount of CD86+HLA-DR + cells reached 77.64 ± 3.11% and CD83+ cells – 28.60 ± 2.79% before DC vaccine treatment and they are quite suitable for IT. Tumor antigens derived from the tumor cell line MDA-MB-231 contributed it possible to obtain sufficient mature DC with CD86 and HLA-DR high expression. The HLA-ABC expression was increased during DC vaccine immunotherapy confirming the enhancement of antigen cross-presentation. The number of HLA-ABC+ cells before IT reached 88.43 ± 3.34% compared to 92.99 ± 0.79% after IT. The maturity of DCs was increased after each subsequent stage of immunotherapy, so to achieve the most significant clinical effect of immunotherapy DC-vaccine must be administered at least 4-5 times. In our study, we developed methodological approach to obtaining the construct «DC-tumor antigen» as a natural adjuvant of antitumor vaccine for the treatment of advanced TNBC patients.

triple-negative breast cancer, immunotherapy, dendritic cells and MDA-MB-231 cells

1. Anders CK, Abramson V, Tan T, Dent R. The Evolution of Triple-Negative Breast Cancer: From Biology to Novel Therapeutics. Am Soc Clin Oncol Educ Book. 2016;35:34-42.
2. Claire H, Karandza V, Aktan G. Current treatment landscape for patients with locally recurrent inoperable or metastatic triple negative breast cancer: a systematic literature review. Breast cancer research. 2019;21:143-57.
3.  Bianchini G, Balko JM, Mayer IA, Sanders ME, Gianni L. Triple negative breast cancer: challenges and opportunities of a heterogeneous disease. Nat. Rev. Clin. Oncol. 2016;13:674-90.
4. Dana A, Franzese E, Centonze S, Carlino F, Della Corte CM, Ventriglia J, et al. Triple-negative breast cancers: systematic review of the literature on molecular and clinical features with a focus on treatment with innovative drugs. Curr. Oncol. Rep. 2018;20(10):76.
5. Lu Z, Qiu Y, Lu W, Jiang Y, Wang G. Immunotherapeutic interventions of Triple Negative Breast Cancer. J Transl Med. 2018;16:147.
6. Banerji S, Cibulskis K, Rangel C, Brown KK, Carter SL, Frederick AM, et al. Sequence analysis of mutations and translocations across breast cancer subtypes. Nature. 2012;486:405-9.
7. Wang ZX, Cao JX, Wang M, Li D, Cui YX, Zhang XY, et al. Adoptive cellular immunotherapy for the treatment of patients with breast cancer: a meta-analysis. Cytotherapy. 2014;16:934-45.
8. Melero I, Gaudernack G, Gerritsen W, Huber C, Parmiani G, Scholl S, et al. Therapeutic vaccines for cancer: an overview of clinical trials. Nat. Rev. Clin. Oncol. 2014;11:509-24.
9. Jung N-C, Lee J-H, Chung K-H, Kwak YS, Lim DS. Dendritic cell-based immunotherapy for solid tumor. Translational Oncology. 2018;11:686-90.
10. Saxena M, Bhardwaj N. Re-emergence of Dendritic Cell Vaccines for Cancer Treatment. Trends Cancer. 2018;4(2):119-37.
11. Gardner A, Ruffell B. Dendritic cells and cancer immunity. Trends Immunol. 2016;37(12):855-65.
12. Rojas-Sepúlveda D, Tittarelli A, Gleisner MA, Ávalos I, Pereda C, Gallegos I, et al. Tumor lysate-based vaccines: on the road to immunotherapy for gallbladder cancer. Cancer Immunol Immunother. 2018;67(12):1897-910.
13. Flores I, Hevia D, Tittarelli A, Soto D, Rojas-Sepúlveda D, Pereda C, et al. Dendritic cells loaded with heats hock-conditioned ovarian epithelial carcinoma cell lysates elicit T cell-dependent antitumor immune responses in vitro. J Immunol Res. 2019;2019:9631515.
14. Brown M-J, Bahsoun S, Morris MA, Akam EC. Determining conditions for successful culture of multi-cellular 3D tumour spheroids to investigate the effect of mesenchymal stem cells on breast cancer cell invasiveness. Bioengineering. 2019;6:101.
15. Khranovska N, Skachkova O, Sovenko V, Sydor P, Inomistova M, Melnyk V. Phenotypic and functional properties of generated dendritic cells in lung cancer patients. Cell and OrganTransplantology. 2016;4(2):162-6.
16. Farkas AM, Marvel D, Finn OJ. Antigen choice determines vaccine-induced generation of immunogenic versus tolerogenic dendritic cells that are marked by differential expression of pancreatic enzymes. J Immunol. 2013;190:19-27.
17. Gustafsson K, Junevik K, Werlenius O, Holmgren S, Karlsson-Parra A, Andersson P. Tumour-loaded α-type 1-polarized dendritic cells from patients with chronic lymphocytic leukaemia produce a superior NK-, NKT- and CD8+ T cell-attracting chemokine profile. Scand J Immunol. 2011;74:18-26.
18. Park J, Gerber MH, Babensee JE. Phenotype and polarization of autologous t cells by biomaterial-treated dendritic cells. J Biomed Mater Res A. 2015;103(1):170-84.
19. Kalinski P, Urban J, Narang R, Berk E, Wieckowski E, Muthuswamy R. Dendritic cell-based therapeutic cancer vaccines: what we have and what we need. Future Oncology. 2009;5(3):379-90.
20. Dudek A, Marti S, Garg A, Agostinis P. Immature, semi-mature, and fully mature dendritic cells: toward a DC-cancer cells interface that augments anticancer immunity. Frontiers In Immunology. 2013;4:40-53.
21. Richter C, Thieme S, Bandoła J. Generation of inducible immortalized dendritic cells with proper immune function in vitro and in vivo. PLoS One. 2013;8(4):62621.
22. Joffre OP, Segura E, Savina A, Amigorena S. Cross-presentation by dendritic cells. Nat Rev Immunol. 2012;12:557-69.
23. Roche PA, Furuta K. The ins and outs of MHC class II-mediated antigen processing and presentation. Nature Reviews Immunology. 2015;34:1-14.

«Bulletin of problems biology and medicine» Issue 2 (156), 2020 year, 124-128 pages, index UDK 616-006.484.04:615.277.3

10.29254/2077-4214-2020-2-156-124-128