M2c-like macrophage associated immunomodulatory function on CD44/CD24 phenotype in triple negative breast cancer (TNBC) subgroups


Etgü O., Bal S. H., Dündar F., Yağcıoğlu B., Özalp E., Oral H. B., ...More

7TH EUROPEAN CONGRESS OF IMMUNOLOGY (ECI2024), Dublin, Ireland, 1 - 04 September 2024, pp.1356

  • Publication Type: Conference Paper / Summary Text
  • City: Dublin
  • Country: Ireland
  • Page Numbers: pp.1356
  • Bursa Uludag University Affiliated: Yes

Abstract

Objective: CD44+/CD24- breast cancer stem cell-like cells are the dominant subgroup of triple negative breast cancer(TNBC) but we have limited understanding of their immunomodulatory effect. This study aims to evaluate M2c-, M2- and M0-like monocyte/macrophage dependent immunofunctional difference on T cells in TNBC and their effect on CD44/CD24 associated phenotype.

Material and method: THP-1 (monocytic), phorbol 12-myristate 13-acetate(PMA)-Induced THP-1(M0/macrophage),M2clike THP-1(CD169+CD163+CD206dim) cells differentiation status were confirmed by NGS and flow cytometry. MDA-MB-231 or MDA-MB-468 triple negative breast cancer (TNBC) cells co-cultured with monocyte/macrophage-like cells were purified with FACS according to their CD44/CD24 expression status. To measure these myleoid cells primed TNBC cells were cocultured with PBMCs obtained from healthy donors and CD4+ or CD8+ T cell proliferation was tested with CFSE dilution by flow cytometry. CD80,CD86,PD-L1 and PD-L2 expression on cancer cells and immune cells were tested by flow cytometry for costimulation. Purified different CD44/CD24 cell subpopulation were plated for scratch assays and analyzed with ImageJ.

Results and discussion: CD44+/CD24- subpopulation collected from M2c-like THP1cells/cancer cells co-culture condition, displayed less metastatic and more immunomodulatory capacity than THP-1 and PMA-Induced THP-1 conditions. In addition,CD163,CD169 vs CD14 expression were decreased on M2c-like THP-1 cells(CD44+/CD24- cancer cells primed).CD44+CD24+ population was decreased in THP-1 derived monocytic/macrophagelike cells coculture.In co-cultures between MDA-MB-468 cell lines and THP-1 derived monocytic/macrophage-like cells, especially CD44+CD24+ population was decreased.CD4+ or CD8+ T cells proliferation were affected to monocyte/macrophage-like cells primed CD44/CD24 sub-populations.In this study,CD44/CD24 phenotype in TNBC and their effects on monocytic/macrophage cell activation polarization were evaluated.In addition, the metastatic character and T cell responses of this interaction were detected,and the first preliminary data that could help immunotherapy approaches.

Keywords: CD4, CD8, T cells, CD44, CD24 Monocyte, Macrophage