Retroviral-mediated IL-12 gene transduction into human CD34+ cell-derived dendritic cells

  • Authors:
    • Y. Akiyama
    • K. Maruyama
    • M. Watanabe
    • K. Yamaguchi
  • View Affiliations

  • Published online on: September 1, 2002     https://doi.org/10.3892/ijo.21.3.509
  • Pages: 509-514
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Abstract

Genetically modified dendritic cells (DCs) with Th1 type cytokine genes are useful for activating anti-tumor immune response. We made human interleukin (IL)-12 p70 gene-transduced DCs generated from CD34+ progenitor cells using a retrovirus system and investigated the function of IL-12-producing DCs. We used the pMX retroviral vector and made cytokine gene-containing viral vectors referred to as GFP pMX and hIL-12 pMX. Supernatants from BOSC23 cells transfected with GFP pMX and hIL-12 pMX were harvested and used for transfection of DC. Cord blood CD34+ cells were incubated with supernatants containing retrovirus for 48 h with cytokines such as IL-3, IL-6, SCF, Flt3 ligand (FL), bFGF and IGF-I. The cells were cultured for 12 days in the presence of GM-CSF, SCF, FL, IL-4 and TNF-α to get mature DC-enriched population. Analysis of surface marker on DCs and allogeneic MLR assay were also performed. After a 14-day culture, 60-70% of cultured CD34+ cells were DC marker (CD1a, DEC205) positive. The IL-12 p70 protein levels in supernatant of DC-GFP and DC-hIL-12 were 0.2 ng/ml and 53 ng/ml/5x105 DCs for 72 h, respectively. The addition of CH296 fibronectin fragment (FN) increased 3-fold IL-12 gene transduction efficiency into DCs. MLR assay showed that IL-12-producing DC exhibited more potent T cell growth-stimulating activity compared with GFP-DC. These results suggested that genetically modified CD34+ cell-derived DCs with human IL-12 gene are fully efficient in T cell priming, and could be a good tool for effective cancer immunotherapy.

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September 2002
Volume 21 Issue 3

Print ISSN: 1019-6439
Online ISSN:1791-2423

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Spandidos Publications style
Akiyama Y, Maruyama K, Watanabe M and Yamaguchi K: Retroviral-mediated IL-12 gene transduction into human CD34+ cell-derived dendritic cells. Int J Oncol 21: 509-514, 2002
APA
Akiyama, Y., Maruyama, K., Watanabe, M., & Yamaguchi, K. (2002). Retroviral-mediated IL-12 gene transduction into human CD34+ cell-derived dendritic cells. International Journal of Oncology, 21, 509-514. https://doi.org/10.3892/ijo.21.3.509
MLA
Akiyama, Y., Maruyama, K., Watanabe, M., Yamaguchi, K."Retroviral-mediated IL-12 gene transduction into human CD34+ cell-derived dendritic cells". International Journal of Oncology 21.3 (2002): 509-514.
Chicago
Akiyama, Y., Maruyama, K., Watanabe, M., Yamaguchi, K."Retroviral-mediated IL-12 gene transduction into human CD34+ cell-derived dendritic cells". International Journal of Oncology 21, no. 3 (2002): 509-514. https://doi.org/10.3892/ijo.21.3.509