open access

Vol 49, No 3 (2011)
Original paper
Submitted: 2012-01-05
Published online: 2011-10-28
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Induction of monocyte antitumor response by human cancer cells transduced with TNF-GFP fusion gene: possible implications for immunotherapy of cancer

Jerzy Więckiewicz, Bożenna Mytar, Rafał Szatanek, Kazimierz Węglarczyk, Jarosław Baran
DOI: 10.5603/FHC.2011.0072
·
Folia Histochem Cytobiol 2011;49(3):512-520.

open access

Vol 49, No 3 (2011)
ORIGINAL PAPERS
Submitted: 2012-01-05
Published online: 2011-10-28

Abstract

This study was undertaken to determine how human pancreatic cancer (HPC-4) cells transduced with the TNF-GFP fusion gene (TLG) alter the antitumor response of human monocytes in vitro and whether they could act as an antitumor vaccine. In our model, HPC-4 cells were transduced with retroviral vector harboring TLG gene and designated as HPC-4TLG. The TLG protein expression was confirmed by Western blot and flow cytometry analysis. Monocytes were co-cultured with transduced and control HPC-4 cells. The secretion of TNF, IL-10 and IL-12 was measured by ELISA. The cytotoxicity of monocytes against HPC-4 cells was determined by MTT test. The results show that the HPC-4TLG cells expressed membrane-bound, intracellular and secretory TLG protein. When cultured with HPC-4TLG cells, monocytes released a higher amount of TNF, but IL-10 and IL-12 secretion was inhibited. The pre-exposure of monocytes to HPC-4TLG, but not to HPC-4, cells did not decrease TNF nor increase IL-10 production, thus not leading to monocyte deactivation. Also, the antitumor cytotoxicity of monocytes stimulated with HPC-4TLG was not downregulated, which occurred when non-transduced HPC-4 cells were used. In conclusion, compared to parental HPC-4 cells, TLG gene transduced HPC-4 cells induced stronger antitumor response of monocytes in vitro and prevented deactivation of monocytes. (Folia Histochemica et Cytobiologica 2011, Vol. 49, No. 3, 512–520)

Abstract

This study was undertaken to determine how human pancreatic cancer (HPC-4) cells transduced with the TNF-GFP fusion gene (TLG) alter the antitumor response of human monocytes in vitro and whether they could act as an antitumor vaccine. In our model, HPC-4 cells were transduced with retroviral vector harboring TLG gene and designated as HPC-4TLG. The TLG protein expression was confirmed by Western blot and flow cytometry analysis. Monocytes were co-cultured with transduced and control HPC-4 cells. The secretion of TNF, IL-10 and IL-12 was measured by ELISA. The cytotoxicity of monocytes against HPC-4 cells was determined by MTT test. The results show that the HPC-4TLG cells expressed membrane-bound, intracellular and secretory TLG protein. When cultured with HPC-4TLG cells, monocytes released a higher amount of TNF, but IL-10 and IL-12 secretion was inhibited. The pre-exposure of monocytes to HPC-4TLG, but not to HPC-4, cells did not decrease TNF nor increase IL-10 production, thus not leading to monocyte deactivation. Also, the antitumor cytotoxicity of monocytes stimulated with HPC-4TLG was not downregulated, which occurred when non-transduced HPC-4 cells were used. In conclusion, compared to parental HPC-4 cells, TLG gene transduced HPC-4 cells induced stronger antitumor response of monocytes in vitro and prevented deactivation of monocytes. (Folia Histochemica et Cytobiologica 2011, Vol. 49, No. 3, 512–520)
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Keywords

fusion gene; monocytes; tumor cells; tumor necrosis factor

About this article
Title

Induction of monocyte antitumor response by human cancer cells transduced with TNF-GFP fusion gene: possible implications for immunotherapy of cancer

Journal

Folia Histochemica et Cytobiologica

Issue

Vol 49, No 3 (2011)

Article type

Original paper

Pages

512-520

Published online

2011-10-28

DOI

10.5603/FHC.2011.0072

Bibliographic record

Folia Histochem Cytobiol 2011;49(3):512-520.

Keywords

fusion gene
monocytes
tumor cells
tumor necrosis factor

Authors

Jerzy Więckiewicz
Bożenna Mytar
Rafał Szatanek
Kazimierz Węglarczyk
Jarosław Baran

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