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IL-15 and IL-2 increase Cetuximab-mediated cellular cytotoxicity against triple negative breast cancer cell lines
M P Roberti1, M M Barrio, A I Bravo
1Centro de Investigaciones Oncológicas, Fundación Cáncer and Instituto Alexander Fleming, Cramer 1180 (1426), Buenos Aires, Argentina.
Abstract:
Triple negative breast cancer (TNBC) patients are not likely to benefit from anti-estrogen or anti-HER2 therapy and this phenotype is associated with a more aggressive clinical course and worse clinical outcome. Taking into account the limited treatment possibilities in TNBC, the aim of the present work was to study a potential therapy based on Cetuximab-mediated immune activity by natural killer (NK) cells. We performed in vitro studies on human breast cancer (BC) cell lines, IIB-BR-G, and the in vivo metastatic variant IIB-BR-G MT. The immunohistochemical analysis showed a TNBC phenotype with high but different levels of EGFR expression on each cell line, measured by flow cytometry. DNA sequencing showed that both cell lines have a mutated K-RAS status, 38 G > A at codon 13. Consequently, Cetuximab did not inhibit cellular proliferation or induce apoptosis. We investigated if Cetuximab could trigger immune mechanisms, and we determined that both cell lines treated with 1 μg/ml Cetuximab were susceptible to antibody dependent cellular cytotoxicity (ADCC), mediated by peripheral blood mononuclear cells (PBMC). At 50:1 effector:target ratio, lytic activity was 34 ± 2% against IIB-BR-G and 27 ± 6% against IIB-BR-G MT cells. PBMC pretreatment with IL-2 allowed reaching 65 ± 3% of Cetuximab-mediated ADCC against IIB-BR-G and 63 ± 6.5% against IIB-BR-G MT. Furthermore, IL-15 pretreatment increased the ADCC up to 71 ± 3% in IIB-BR-G and 79 ± 3.5% in IIB-BR-G MT. We suggest that NK cells are the effectors present in PBMC since they were able to induce ADCC at lower effector:target ratios. Besides, IL-2- and mainly IL-15-induced upregulation of NK activating receptors CD16 and NKG2D and enhanced IFN-γ production. EGFR-expressing TNBC could be killed by Cetuximab-mediated ADCC at clinically achievable concentrations. IL-15 could advantageously replace IL-2 in most of its immunologic activities, stimulating the ability to produce IFN-γ, and paralleling the up-regulation of activating receptors.
Insights
Cetuximab triggers natural killer (NK) cell immune activity against triple-negative breast cancer (TNBC) via antibody-dependent cellular cytotoxicity (ADCC). Interleukin-15 (IL-15) significantly enhances this NK cell-mediated ADCC response in EGFR-expressing TNBC.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- Triple-negative breast cancer (TNBC) lacks targeted therapies like anti-estrogen or anti-HER2 treatments, presenting a significant clinical challenge.
- TNBC is characterized by aggressive behavior and poorer patient outcomes, necessitating novel therapeutic strategies.
- EGFR expression is present in TNBC, offering a potential target for immunotherapeutic interventions.
Purpose of the Study:
- To investigate the potential of Cetuximab-mediated immune activity by natural killer (NK) cells as a therapy for triple-negative breast cancer (TNBC).
- To evaluate the efficacy of Cetuximab in triggering antibody-dependent cellular cytotoxicity (ADCC) in EGFR-expressing TNBC cell lines.
- To assess the role of cytokines Interleukin-2 (IL-2) and Interleukin-15 (IL-15) in enhancing Cetuximab-mediated ADCC.
Main Methods:
- In vitro studies using human TNBC cell lines (IIB-BR-G and IIB-BR-G MT) with confirmed EGFR expression and mutated K-RAS status.
- Assessment of Cetuximab's effect on cellular proliferation and apoptosis.
- Measurement of Cetuximab-mediated ADCC by peripheral blood mononuclear cells (PBMCs) at various effector:target ratios.
- Evaluation of cytokine (IL-2, IL-15) pretreatment effects on PBMC-mediated ADCC and NK cell activation markers (CD16, NKG2D) and IFN-γ production.
Main Results:
- Cetuximab did not inhibit proliferation or induce apoptosis in the studied TNBC cell lines.
- Both TNBC cell lines exhibited susceptibility to Cetuximab-mediated ADCC by PBMCs.
- IL-2 and IL-15 pretreatment significantly enhanced Cetuximab-mediated ADCC, with IL-15 showing superior efficacy.
- IL-15 upregulated NK activating receptors (CD16, NKG2D) and enhanced IFN-γ production, suggesting NK cells as key mediators.
Conclusions:
- Cetuximab can induce ADCC in EGFR-expressing TNBC, offering a potential immunotherapeutic approach.
- IL-15 is a potent enhancer of Cetuximab-mediated ADCC in TNBC, potentially improving treatment outcomes.
- NK cells are the primary effector cells in Cetuximab-mediated ADCC, and their activity can be significantly boosted by IL-15.
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