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Generation of interleukin-13 receptor alpha2 antigen expressing modified vaccinia ankara recombinant virus for potential cancer immunotherapy

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Genetically modified recombinant poxviruses have shown promise in preclinical models of cancer immunotherapy due to their ability to induce effective cell-mediated immunity against target tumor-associated antigens (TAA). One such vector, recombinant Modified Vaccinia Ankara (MVA), is capable of expressing foreign genes in infected host cells. MVA is replication restricted in most mammalian cells exemplifying a unique safety profile. We have demonstrated that the interleukin-13 receptor α2 (IL-13Rα2) is selectively expressed in various solid tumors but not in normal tissues making it a promising TAA. Prophylactic and therapeutic vaccination with a plasmid vector expressing IL-13Rα2 caused only partial regression of established tumors [1], suggesting that host immune responses against IL-13Rα2 needed further enhancement. Thus, we constructed a recombinant MVA (rMVA-IL13Rα2) expressing both IL-13Rα2 and a green fluorescent protein (GFP) reporter gene. Purified virus titration by immunostaining using anti-vaccinia antibody and anti-IL-13Rα2 antibody confirmed the identity and purity of the recombinant MVA. Western Blot analysis showed the presence of IL-13Rα2 protein (65 kDa). Flow cytometric analysis of IL-13Rα2 negative T98G glioma cells infected with rMVA-IL13Rα2 virus (T98G-IL13Rα2) demonstrated surface expression of IL-13Rα2, indicating the infectivity potential of the recombinant virus. Incubation of T98G-IL13Rα2 cells with varying concentrations (0-100 ng/ml) of IL13-PE (interleukin-13 fused to truncated Pseudomonas exotoxin [2] resulted in depletion of GFP+ T98G-IL13Rα2 cells in a concentration-dependent manner. Higher concentrations of IL13-PE (10-1000 ng/ml) also inhibited the protein synthesis in T98G-IL13Rα2 compared to cells infected with control pLW44-MVA. We further observed that IL13-PE treatment of rMVA-IL13Rα2 infected chicken fibroblast, DF-1 cells led to a reduction in virus titer compared to untreated cells. These results indicate that rMVA-IL13Rα2 virus can successfully infect mammalian cells and express IL-13Rα2 in a biologically active form on the cell surface. The immunization studies of rMVA-IL13Rα2 are ongoing in a syngeneic mouse model of metastatic breast carcinoma. Based on in vitro results, we expect the rMVA-IL13Rα2 to be a useful agent in tumor immunotherapy as a vaccine alone and in combination with other therapeutic agents to eradicate metastatic tumors.


  1. 1.

    Nakashima H, Terabe M, Husain SR, Puri RK: A novel combination immunotherapy for cancer by IL-13Ra2-targeted DNA vaccine and immunotoxin in murine tumor models. J Immunol. 2011, 187: 4935-4946. 10.4049/jimmunol.1102095.

  2. 2.

    Husain SR, Puri RK: Interleukin-13 receptor-directed cytotoxin for malignant glioma therapy: from bench to bedside. J Neuro-Oncol. 2003, 65: 37-48. 10.1023/A:1026242432647.

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Correspondence to Raj K Puri.

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Sato, Y., Vatsan, R., Joshi, B.H. et al. Generation of interleukin-13 receptor alpha2 antigen expressing modified vaccinia ankara recombinant virus for potential cancer immunotherapy. j. immunotherapy cancer 2, P58 (2014) doi:10.1186/2051-1426-2-S3-P58

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  • Green Fluorescent Protein
  • Vaccinia
  • Recombinant Virus
  • Cancer Immunotherapy
  • Infected Host Cell