Human anti-CCR4 minibody gene transfer for the treatment of cutaneous T-cell lymphoma

PLoS One. 2012;7(9):e44455. doi: 10.1371/journal.pone.0044455. Epub 2012 Sep 4.

Abstract

Background: Although several therapeutic options have become available for patients with Cutaneous T-cell Lymphoma (CTCL), no therapy has been curative. Recent studies have demonstrated that CTCL cells overexpress the CC chemokine receptor 4 (CCR4).

Methodology/principal findings: In this study, a xenograft model of CTCL was established and a recombinant adeno-associated viral serotype 8 (AAV8) vector expressing a humanized single-chain variable fragment (scFv)-Fc fusion (scFvFc or "minibody") of anti-CCR4 monoclonal antibody (mAb) h1567 was evaluated for curative treatment. Human CCR4+ tumor-bearing mice treated once with intravenous infusion of AAV8 virions encoding the h1567 (AAV8-h1567) minibody showed anti-tumor activity in vivo and increased survival. The AAV8-h1567 minibody notably increased the number of tumor-infiltrating Ly-6G+ FcγRIIIa(CD16A)+ murine neutrophils in the tumor xenografts over that of AAV8-control minibody treated mice. Furthermore, in CCR4+ tumor-bearing mice co-treated with AAV8-h1567 minibody and infused with human peripheral blood mononuclear cells (PBMCs), marked tumor infiltration of human CD16A+ CD56+ NK cells was observed. The h1567 minibody also induced in vitro ADCC activity through both mouse neutrophils and human NK cells.

Conclusions/significance: Overall, our data demonstrate that the in vivo anti-tumor activity of h1567 minibody is mediated, at least in part, through CD16A+ immune effector cell ADCC mechanisms. These data further demonstrate the utility of the AAV-minibody gene transfer system in the rapid evaluation of candidate anti-tumor mAbs and the potency of h1567 as a potential novel therapy for CTCL.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Analysis of Variance
  • Animals
  • Antibodies, Monoclonal / genetics
  • Antibodies, Monoclonal / therapeutic use*
  • Blotting, Western
  • DNA Primers / genetics
  • Dependovirus / genetics
  • Flow Cytometry
  • Genetic Therapy / methods*
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Genetic Vectors / metabolism
  • Humans
  • Image Processing, Computer-Assisted
  • Immunoglobulin Fc Fragments / genetics
  • Immunoglobulin Fc Fragments / metabolism*
  • Immunohistochemistry
  • Lymphoma, T-Cell, Cutaneous / genetics
  • Lymphoma, T-Cell, Cutaneous / therapy*
  • Mice
  • Mice, SCID
  • Real-Time Polymerase Chain Reaction
  • Receptors, CCR4 / metabolism*
  • Single-Chain Antibodies / genetics
  • Single-Chain Antibodies / metabolism*
  • Transduction, Genetic

Substances

  • Antibodies, Monoclonal
  • CCR4 protein, human
  • DNA Primers
  • Immunoglobulin Fc Fragments
  • Receptors, CCR4
  • Single-Chain Antibodies