TY - JOUR
T1 - IL-15 and IL-2 increase Cetuximab-mediated cellular cytotoxicity against triple negative breast cancer cell lines expressing EGFR
AU - Roberti, M. P.
AU - Barrio, M. M.
AU - Bravo, A. I.
AU - Rocca, Y. S.
AU - Arriaga, J. M.
AU - Bianchini, M.
AU - Mordoh, J.
AU - Levy, E. M.
N1 - Funding Information:
Acknowledgments This work was supported with funds from Fun-dación Sales, Fundación P. Mosoteguy, Fundación Cáncer (FUCA), Agencia Nacional de Promoción Científica y Tecnológica (ANPCyT) and Fundación María Calderón de la Barca. M. Bianchini, M.M. Barrio, J. Mordoh, and E.M. Levy are members of CONICET. We are grateful to Dr. Norberto W. Zwirner for kindly providing us IL-15 and Biochemist Fernanda Reynoso for performing PCR for K-RAS analysis and to Ms. María Luisa Poljak for revising this manuscript. We also thank the Servicio de Hemoterapia del Instituto Médico Espe-cializado A. Fleming for providing healthy donors blood samples.
PY - 2011/11
Y1 - 2011/11
N2 - 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 IIBBR- G MT. Furthermore, IL-15 pretreatment increased the ADCC up to 71 ± 3% in IIB-BR-G and 79 ± 3.5% in IIBBR- 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. EGFRexpressing 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-c, and paralleling the up-regulation of activating receptors.
AB - 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 IIBBR- G MT. Furthermore, IL-15 pretreatment increased the ADCC up to 71 ± 3% in IIB-BR-G and 79 ± 3.5% in IIBBR- 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. EGFRexpressing 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-c, and paralleling the up-regulation of activating receptors.
KW - Cetuximab-mediated cytotoxicity
KW - IL-2 and IL-15
KW - K-RAS status
KW - Triple negative breast cancer
UR - https://www.scopus.com/pages/publications/82955165645
U2 - 10.1007/s10549-011-1360-2
DO - 10.1007/s10549-011-1360-2
M3 - Article
C2 - 21308409
AN - SCOPUS:82955165645
SN - 0167-6806
VL - 130
SP - 465
EP - 475
JO - Breast Cancer Research and Treatment
JF - Breast Cancer Research and Treatment
IS - 2
ER -