GM-CSF enhances 3F8 monoclonal antibody-dependent cellular cytotoxicity against human melanoma and neuroblastoma

Abstract

3F8 is a murine monoclonal IgG3 antibody specific for the tumor- associated antigen ganglioside GD2. Previous in vitro studies suggest that tumor regressions observed in a phase I clinical trial of 3F8 may be attributable to complement activation by 3F8 and to 3F8-dependent cellular cytotoxicity (ADCC) with lymphocytes. We now describe 3F8- mediated ADCC of GD2-positive tumor targets (melanoma and neuroblastoma) with human granulocytes and report that recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) enhanced this phenomenon. Cytotoxicity required binding of 3F8 to the low-affinity Fc receptor type III (CD16) on the granulocytes and was poor with tumor-binding monoclonal antibodies of other immunoglobulin (ie, non-IgG3) subclasses. GM-CSF (2 to 20 ng/mL) increased ADCC by 93% to 267% at limiting dilutions of 3F8 (1 microgram/mL). With most GD2- positive cell lines tested, this effect translated into a tenfold or greater augmentation in 3F8 efficiency at mediating ADCC. Comparable enhancement occurred whether GM-CSF was present in the ADCC assay or granulocytes were incubated with GM-CSF and washed before the assay. Nonoxidative mechanisms may be important for ADCC since 3F8 mediated ADCC with granulocytes from two children with chronic granulomatous disease; this cytotoxicity was also enhanced by GM-CSF. Since GM-CSF induces a neutrophilia in patients, our data suggest that this cytokine may have the potential of amplifying 3F8 antitumor activity in patients by increasing effector cell numbers and by priming granulocytes for greater cytotoxicity.

Tumour Immunogenicity, Antigen Presentation, and Immunological Barriers in Cancer Immunotherapy

“…The inflammatory environment also strongly contributes to their differentiation and acquisition of their suppressive activities. Granulocyte monocyte colony stimulating factor (GM-CSF) produced by cancer cells strongly contributes to MDSC differentiation [296,298,303,304], which would explain why the use of GM-CSF in human cancer immunotherapy has mixed responses [305][306][307][308][309][310][311].…”

Immunotherapeutic Strategies for Neuroblastoma: Present, Past and Future

“…The effectiveness of GD2 Ab is related to different mechanisms as (i) direct induction of cell death, (ii) Fcγ receptor (FcγR)–mediated antibody-dependent cell-mediated cytotoxicity (ADCC) by immune cells such as NK, macrophages and neutrophils and (iii) complement-dependent cytotoxicity (CDC) [ 47 , 48 , 49 , 50 ]. Of note, these mechanisms of cytotoxicity may be highly potentiated by the use of immune stimulating cytokines or adoptive cell therapy.…”

Glycobiology of Neuroblastoma: Impact on Tumor Behavior, Prognosis, and Therapeutic Strategies

“…Its efficacy in treating NB was initially described in 1987 in the report of a Phase 1 trial which included patients with refractory high-risk NB ( 159 ). MAb 3F8 mediates highly efficient antibody-dependent cell-mediated cytotoxicity (ADCC) of NB in the presence of human natural killer (NK) cells and granulocytes in vitro ( 160 – 162 ). Moreover, it induces complement-mediated cytotoxicity (CMC), because NB cells lack decay-accelerating factor CD55 ( 163 ) and homologous restriction factor CD59 ( 164 ).…”

“…Moreover, it induces complement-mediated cytotoxicity (CMC), because NB cells lack decay-accelerating factor CD55 ( 163 ) and homologous restriction factor CD59 ( 164 ). IL-2 and GM-CSF were shown to enhance ADCC in vitro by activating cytotoxic NK cells and neutrophils ( 160 , 165 , 166 ). When combined with the cytokine GM-CSF and 13-cis-retinoic acid, 3F8 induced >60% long-term survival among high-risk patients with metastatic disease, treated at first remission ( 167 ).…”