Antibody-dependent cell-mediated cytotoxicity can be induced by MUC1 peptide vaccination of breast cancer patients.
Human polymorphic epithelial mucin (PEM, MUC1) is a high molecular weight transmembrane glycoprotein expressed on the apical cell surface of glandular epithelium and is over-expressed and hypo-glycosylated in adenocarcinomas. The extracellular part of the molecule consists mainly of a variable number of 20 amino acid repeats that contain cryptic epitopes exposed in malignancy. The objective of our study was to determine whether humanized MUC1 MAbs and Abs induced by vaccination of breast cancer patients with MUC1 peptides can effect an antibody-dependent cell-mediated cytotoxicity (ADCC). An in vitro assay has been set up in which the breast tumor cell line ZR-75-1 is used as target and PBMC of healthy donors as effector cells. Different target and effector cells, as well as various MUC1 MAbs were tested to optimize the efficacy of the in vitro assay. The humanized MAb HuHMFG-1, which recognizes the PDTR sequence in the MUC1 tandem repeat, induced a strong cell-mediated cytotoxicity. Nine MUC1-expressing tumor cell lines, including 3 bone marrow-derived cell lines, as well as 2 MUC1-transfected cell lines were susceptible to different extent to MUC1 Ab-dependent killing. Large variations in the killing capacity of PBMC from healthy donors were found. The NK cells were the essential effector cells for the MUC1 Ab-dependent killing. Plasma samples with induced high levels of MUC1 Ab were obtained from breast cancer patients repeatedly immunized with a KLH-conjugated 33-mer or 106-mer MUC1 tandem repeat. Pre- and post-vaccinated plasma samples of these patients were compared in the ADCC assay and it could be clearly demonstrated that the induced MUC1 Abs can effect tumor cell killing. MUC1 Ab-dependent cell-mediated tumor cell killing may occur in vivo and the ADCC assay can be applied to monitor MUC1 vaccination trials.
Snijdewint
F G
FG
Department of Obstetrics and Gynaecology, Academic Hospital Vrije Universiteit, Amsterdam, The Netherlands. snijdewint@azu.nl
von Mensdorff-Pouilly
S
S
Karuntu-Wanamarta
A H
AH
Verstraeten
A A
AA
Livingston
P O
PO
Hilgers
J
J
Kenemans
P
P
D000595:Amino Acid Sequence; D000920:Antibody-Dependent Cell Cytotoxicity; D001323:Autoantibodies / Q000276:immunology; D001854:Bone Marrow Cells / Q000473:pathology; D001943:Breast Neoplasms / Q000097:blood / Q000276:immunology* / Q000473:pathology / Q000628:therapy; D019496:Cancer Vaccines / Q000276:immunology* / Q000627:therapeutic use; D005260:Female; D006650:Histocompatibility Testing; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood; D007075:Immunoglobulin M / Q000097:blood; D007694:Killer Cells, Natural / Q000276:immunology*; D008969:Molecular Sequence Data; D018396:Mucin-1 / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D009367:Neoplasm Staging; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D011994:Recombinant Proteins / Q000276:immunology; D012016:Reference Values; D020449:Repetitive Sequences, Amino Acid; D014162:Transfection; D014407:Tumor Cells, Cultured
D001323:Autoantibodies; D019496:Cancer Vaccines; D007074:Immunoglobulin G; D007075:Immunoglobulin M; C116055:MUC1 tandem repeat peptide; D018396:Mucin-1; D010446:Peptide Fragments; D011994:Recombinant Proteins