Vol 49, No 3 (2011)
Original paper
Submitted: 2012-01-05
Published online: 2011-10-28
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.
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)
Keywords
fusion gene; monocytes; tumor cells; tumor necrosis factor
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
Page views
1892
Article views/downloads
1814
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