Construction and characterization of a recombinant adenoviral vector expressing human interleukin-12

Jian Qiao, Shu Hsia Chen, Khiem B. Pham-Nguyen, John Mandeli, Savio L.C. Woo

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Interleukin-12 (IL-12) is a 70-kDa heterodimeric cytokine composed of a 35-kDa subunit (p35) and a 40-kDa subunit (p40). We have demonstrated previously that intratumoral delivery of a recombinant adenoviral vector expressing the mouse IL-12 gene significantly prolongs the survival time of mice with metastatic colon carcinoma in the liver. We now report the molecular cloning of cDNA for both subunits of human IL-12 (hIL-12) in a recombinant adenoviral vector in which the p40 and p35 subunits are linked and coexpressed using the encephalomyocarditis virus internal ribosome entry site. The recombinant adenoviral vector was used to transduce human tumor cell lines, and the presence of hIL-12 in the conditioned media was illustrated by enzyme-linked immunosorbent assay. The biological activity of hIL-12 in the conditioned media was also demonstrated in vitro through its ability to induce interferon-γ production from peripheral blood mononuclear cells (PBMCs), to stimulate PBMC proliferation, and to enhance natural killer activity from normal human PBMCs to lyse natural killer-sensitive K562 target cells. The results of these studies support the application of this recombinant adenoviral vector construct as an efficient gene delivery vehicle in phase I/II clinical studies of hIL-12 gene therapy for cancer.

Original languageEnglish (US)
Pages (from-to)373-379
Number of pages7
JournalCancer Gene Therapy
Volume6
Issue number4
StatePublished - Dec 1 1999

Fingerprint

Interleukin-12
Blood Cells
Conditioned Culture Medium
Human Activities
Encephalomyocarditis virus
K562 Cells
Molecular Cloning
Tumor Cell Line
Genetic Therapy
Interferons
Genes
Colon
Complementary DNA
Enzyme-Linked Immunosorbent Assay
Cell Proliferation
Cytokines
Carcinoma
Survival
Liver
Neoplasms

Keywords

  • Antitumor immunity
  • Gene therapy
  • Human interleukin-12
  • Recombinant adenoviral vector

ASJC Scopus subject areas

  • Molecular Medicine
  • Molecular Biology
  • Cancer Research

Cite this

Qiao, J., Chen, S. H., Pham-Nguyen, K. B., Mandeli, J., & Woo, S. L. C. (1999). Construction and characterization of a recombinant adenoviral vector expressing human interleukin-12. Cancer Gene Therapy, 6(4), 373-379.

Construction and characterization of a recombinant adenoviral vector expressing human interleukin-12. / Qiao, Jian; Chen, Shu Hsia; Pham-Nguyen, Khiem B.; Mandeli, John; Woo, Savio L.C.

In: Cancer Gene Therapy, Vol. 6, No. 4, 01.12.1999, p. 373-379.

Research output: Contribution to journalArticle

Qiao, J, Chen, SH, Pham-Nguyen, KB, Mandeli, J & Woo, SLC 1999, 'Construction and characterization of a recombinant adenoviral vector expressing human interleukin-12', Cancer Gene Therapy, vol. 6, no. 4, pp. 373-379.
Qiao, Jian ; Chen, Shu Hsia ; Pham-Nguyen, Khiem B. ; Mandeli, John ; Woo, Savio L.C. / Construction and characterization of a recombinant adenoviral vector expressing human interleukin-12. In: Cancer Gene Therapy. 1999 ; Vol. 6, No. 4. pp. 373-379.
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