In vivo characterization of fusion protein comprising of A1 subunit of Shiga toxin and human GM-CSF: Assessment of its immunogenicity and toxicity

Iran Biomed J. 2010 Oct;14(4):136-41.

Abstract

Background: Most cancer cells become resistant to anti-cancer agents. In the last few years, a new approach for targeted therapy of human cancer has been developed using immunotoxins which comprise both the cell targeting and the cell killing moieties.

Methods: In the present study, the recombinant Shiga toxin A1 subunit fused to human granulocyte-macrophage colony stimulating factor (A1-GM-CSF), previously produced in E. coli, was further characterized.

Results: The recombinant protein could cause 50% cytotoxicity and induced apoptosis in cells bearing GM-CSF receptors. The non-specific toxicity of the fusion protein was assessed in C57BL/6 and BALB/c mice. No mortality was observed in either group of mice, with different concentration of fusion protein.

Conclusion: The lymphocyte proliferation assay, induction of specific IgG response and a mixed (Th1/Th2) response were observed only in BALB/c mice. The mixed response in BALB/c mice (Th1/Th2) could be explained on the basis of the two components of the fusion protein i.e. A1 and GM-CSF.

Keywords: Cell line; Fusion protein; Toxicity.

MeSH terms

  • Animals
  • Antineoplastic Agents / immunology*
  • Antineoplastic Agents / toxicity*
  • Cytotoxicity, Immunologic*
  • Female
  • Granulocyte-Macrophage Colony-Stimulating Factor / genetics
  • Granulocyte-Macrophage Colony-Stimulating Factor / immunology*
  • Granulocyte-Macrophage Colony-Stimulating Factor / toxicity
  • Humans
  • Immunoglobulin G / blood
  • Immunoglobulin G / immunology
  • Immunotoxins / immunology*
  • Immunotoxins / toxicity*
  • Interferon-gamma / analysis
  • Interleukin-4 / analysis
  • K562 Cells
  • Lethal Dose 50
  • Lymphocyte Activation
  • Lymphocytes / immunology
  • Mice
  • Mice, Inbred BALB C
  • Mice, Inbred C57BL
  • Molecular Sequence Data
  • Rabbits
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / immunology*
  • Recombinant Fusion Proteins / toxicity*
  • Shiga Toxin / genetics
  • Shiga Toxin / immunology*
  • Shiga Toxin / toxicity*

Substances

  • Antineoplastic Agents
  • Immunoglobulin G
  • Immunotoxins
  • Recombinant Fusion Proteins
  • Shiga toxin subunit A
  • Interleukin-4
  • Shiga Toxin
  • Interferon-gamma
  • Granulocyte-Macrophage Colony-Stimulating Factor

Associated data

  • GENBANK/EF032873