Differential Response of MDA-MB-231 and MCF-7 Breast Cancer Cells to In Vitro Inhibition with CTLA-4 and PD-1 through Cancer-Immune Cells Modified Interactions
Kamil Grubczak,
Anna Kretowska-Grunwald,
Dawid Groth,
Izabela Poplawska,
Andrzej Eljaszewicz,
Lukasz Bolkun,
Aleksandra Starosz,
Jordan M. Holl,
Marta Mysliwiec,
Joanna Kruszewska,
Marek Z. Wojtukiewicz,
Marcin Moniuszko
Affiliations
Kamil Grubczak
Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Anna Kretowska-Grunwald
Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Dawid Groth
Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Izabela Poplawska
Department of Medical Pathomorphology, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Andrzej Eljaszewicz
Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Lukasz Bolkun
Department of Haematology, Medical University of Bialystok, M. Sklodowskiej-Curie 24A, 15-276 Bialystok, Poland
Aleksandra Starosz
Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Jordan M. Holl
Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Marta Mysliwiec
Department of Oncology, Medical University of Bialystok, Ogrodowa 12, 15-027 Bialystok, Poland
Joanna Kruszewska
Department of Oncology, Medical University of Bialystok, Ogrodowa 12, 15-027 Bialystok, Poland
Marek Z. Wojtukiewicz
Department of Oncology, Medical University of Bialystok, Ogrodowa 12, 15-027 Bialystok, Poland
Marcin Moniuszko
Department of Regenerative Medicine and Immune Regulation, Medical University of Bialystok, Jerzego Waszyngtona 13, 15-269 Bialystok, Poland
Drugs targeting immune checkpoint molecules have been found effective in melanoma, lung cancer, and other malignancies treatment. Recent studies on breast cancer demonstrated the significance of inhibitory anti-CTLA-4 and anti-PD-1 in the regulation of disease progression. However, seemingly the same types of breast cancer do not always respond unambiguously to immunotherapy. Thus, here we set out to analyze the in vitro effects of inhibiting CTLA-4 and PD-1 on interactions between co-cultured lymphocytes and two selected breast adenocarcinoma cell lines. Breast cancer cells were co-cultured with lymphocytes to evaluate the effects of CTLA-4 and PD-1 inhibition. Proliferation, cell cycle, and viability assessment were measured in cancer cells. IFN-gamma, IL-10, perforin, granzyme B production, and CTLA-4 and PD-1 expression were analyzed in lymphocytes. We found that administration of anti-CTLA-4 improved the anti-cancer activity of T cells with reduced proliferation and viability of MDA-MB-231. Lack of response was observed in the context of MCF-7. In addition, differential expression of checkpoint proteins was found between studied cancer cells lines. Inhibition of molecules was followed by IL-10 and IFN-gamma decrease in lymphocytes co-cultured with MDA-MB-231, not demonstrated in reference to MCF-7. Furthermore, CTLA-4 blockage was associated with reduction of CTLA-4+ and PD-1+ lymphocytes in MDA-MB-231, with a significant increase in MCF-7, reduced by anti-PD-1. Altogether, our study revealed that anti-CTLA-4 and anti-PD-1 treatment can improve lymphocytes effects on breast cancer cells. Favorable effects seemed to be related to breast cancer cells features as differential responses were reported. Novel blocking antibodies strategies should be tested for more effective cancer inhibition.
