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Adenovirus-mediated interferon alpha gene transfer induces regional direct cytotoxicity and possible systemic immunity against pancreatic cancer.

Ohashi M, Yoshida K, Kushida M, Miura Y, Ohnami S, Ikarashi Y, Kitade Y, Yoshida T, Aoki K - Br. J. Cancer (2005)

Bottom Line: Here, we dissected and examined the two mechanisms, taking advantage of the fact that IFN-alpha did not show any cross-species activity in its in vivo effect.The IFN-alpha protein concentration was markedly increased in the injected subcutaneous tumour, but leakage of the potent cytokine into the systemic blood circulation was minimal.Moreover, in this case, tumour regression was observed not only for the injected subcutaneous tumours but also for the untreated tumours at distant sites.

View Article: PubMed Central - PubMed

Affiliation: Genetics Division, National Cancer Center Research Institute, Chuo-ku, Tokyo, Japan.

ABSTRACT
We previously demonstrated a characteristically high sensitivity of pancreatic cancer cells to interferon alpha (IFN-alpha) gene transfer, which induced a more prominent growth suppression and cell death in pancreatic cancer cells than in other types of cancers and normal cells. The IFN-alpha protein can exhibit both direct cytotoxicity and indirect immunological antitumour activity. Here, we dissected and examined the two mechanisms, taking advantage of the fact that IFN-alpha did not show any cross-species activity in its in vivo effect. When a human IFN-alpha adenovirus was injected into subcutaneous xenografts of human pancreatic cancer cells in nude mice, tumour growth was significantly suppressed due to cell death in an adenoviral dose-dependent manner. The IFN-alpha protein concentration was markedly increased in the injected subcutaneous tumour, but leakage of the potent cytokine into the systemic blood circulation was minimal. When a mouse IFN-alpha adenovirus was injected into the same subcutaneous tumour system, all mice showed significant tumour inhibition, an effect that was dependent on the indirect antitumour activities of IFN-alpha, notably a stimulation of natural killer cells. Moreover, in this case, tumour regression was observed not only for the injected subcutaneous tumours but also for the untreated tumours at distant sites. This study suggested that a local IFN-alpha gene therapy is a promising therapeutic strategy for pancreatic cancer, due to its dual mechanisms of antitumour activities and lack of significant toxicity.

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Expression of NKG2D ligands on pancreatic cancer cell lines. (A) RT–PCR analyses of NKG2D ligands. Total RNA was extracted from cell lines and after reverse transcription the mRNA of MICA, MICB, ULBP-1, ULBP-2 and ULBP-3 was detected by PCR. (B) Flow cytometric analysis of MICA and MICB. Cells were incubated with anti-human MICA/B monoclonal antibody (filled curve) or isotype control antibody (open curve), and flow cytometry was carried out using a FACScan system.
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fig5: Expression of NKG2D ligands on pancreatic cancer cell lines. (A) RT–PCR analyses of NKG2D ligands. Total RNA was extracted from cell lines and after reverse transcription the mRNA of MICA, MICB, ULBP-1, ULBP-2 and ULBP-3 was detected by PCR. (B) Flow cytometric analysis of MICA and MICB. Cells were incubated with anti-human MICA/B monoclonal antibody (filled curve) or isotype control antibody (open curve), and flow cytometry was carried out using a FACScan system.

Mentions: NK cells are an important element of the innate immune system as they are capable of killing tumour cells. The cytotoxic activity of NK cells is enhanced by IFN-α/β through TRAIL (tumour necrosis factor-related apoptosis-inducing ligand) gene induction (Sato et al, 2001). NKG2D is one of the activating receptors on NK cells, and a number of NKG2D ligands, such as MICA and MICB (major histocompatibility complex class I chain-related) and ULBP (UL16-binding proteins), have been identified (Groh et al, 1999; Bahram, 2000; Kubin et al, 2001; Stephens, 2001; Raulet, 2003; Yokoyama and Plougastel, 2003). It is known that the expression of NKG2D ligands is upregulated in transformed cells, and that the human NKG2D ligands can bind to mouse NKG2D, leading to the activation of mouse NK cells (Kubin et al, 2001). To examine whether the pancreatic cancer cells express the ligands, RT–PCR and flow cytometric analyses were performed in four cell lines (Panc-1, AsPC-1, BxPC-3 and MIAPaCa-2). As shown in Figure 5, all the pancreatic cancer cells examined expressed several kinds of NKG2D ligands, suggesting that the cancer cells are effective targets of NK cells.


Adenovirus-mediated interferon alpha gene transfer induces regional direct cytotoxicity and possible systemic immunity against pancreatic cancer.

Ohashi M, Yoshida K, Kushida M, Miura Y, Ohnami S, Ikarashi Y, Kitade Y, Yoshida T, Aoki K - Br. J. Cancer (2005)

Expression of NKG2D ligands on pancreatic cancer cell lines. (A) RT–PCR analyses of NKG2D ligands. Total RNA was extracted from cell lines and after reverse transcription the mRNA of MICA, MICB, ULBP-1, ULBP-2 and ULBP-3 was detected by PCR. (B) Flow cytometric analysis of MICA and MICB. Cells were incubated with anti-human MICA/B monoclonal antibody (filled curve) or isotype control antibody (open curve), and flow cytometry was carried out using a FACScan system.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2361577&req=5

fig5: Expression of NKG2D ligands on pancreatic cancer cell lines. (A) RT–PCR analyses of NKG2D ligands. Total RNA was extracted from cell lines and after reverse transcription the mRNA of MICA, MICB, ULBP-1, ULBP-2 and ULBP-3 was detected by PCR. (B) Flow cytometric analysis of MICA and MICB. Cells were incubated with anti-human MICA/B monoclonal antibody (filled curve) or isotype control antibody (open curve), and flow cytometry was carried out using a FACScan system.
Mentions: NK cells are an important element of the innate immune system as they are capable of killing tumour cells. The cytotoxic activity of NK cells is enhanced by IFN-α/β through TRAIL (tumour necrosis factor-related apoptosis-inducing ligand) gene induction (Sato et al, 2001). NKG2D is one of the activating receptors on NK cells, and a number of NKG2D ligands, such as MICA and MICB (major histocompatibility complex class I chain-related) and ULBP (UL16-binding proteins), have been identified (Groh et al, 1999; Bahram, 2000; Kubin et al, 2001; Stephens, 2001; Raulet, 2003; Yokoyama and Plougastel, 2003). It is known that the expression of NKG2D ligands is upregulated in transformed cells, and that the human NKG2D ligands can bind to mouse NKG2D, leading to the activation of mouse NK cells (Kubin et al, 2001). To examine whether the pancreatic cancer cells express the ligands, RT–PCR and flow cytometric analyses were performed in four cell lines (Panc-1, AsPC-1, BxPC-3 and MIAPaCa-2). As shown in Figure 5, all the pancreatic cancer cells examined expressed several kinds of NKG2D ligands, suggesting that the cancer cells are effective targets of NK cells.

Bottom Line: Here, we dissected and examined the two mechanisms, taking advantage of the fact that IFN-alpha did not show any cross-species activity in its in vivo effect.The IFN-alpha protein concentration was markedly increased in the injected subcutaneous tumour, but leakage of the potent cytokine into the systemic blood circulation was minimal.Moreover, in this case, tumour regression was observed not only for the injected subcutaneous tumours but also for the untreated tumours at distant sites.

View Article: PubMed Central - PubMed

Affiliation: Genetics Division, National Cancer Center Research Institute, Chuo-ku, Tokyo, Japan.

ABSTRACT
We previously demonstrated a characteristically high sensitivity of pancreatic cancer cells to interferon alpha (IFN-alpha) gene transfer, which induced a more prominent growth suppression and cell death in pancreatic cancer cells than in other types of cancers and normal cells. The IFN-alpha protein can exhibit both direct cytotoxicity and indirect immunological antitumour activity. Here, we dissected and examined the two mechanisms, taking advantage of the fact that IFN-alpha did not show any cross-species activity in its in vivo effect. When a human IFN-alpha adenovirus was injected into subcutaneous xenografts of human pancreatic cancer cells in nude mice, tumour growth was significantly suppressed due to cell death in an adenoviral dose-dependent manner. The IFN-alpha protein concentration was markedly increased in the injected subcutaneous tumour, but leakage of the potent cytokine into the systemic blood circulation was minimal. When a mouse IFN-alpha adenovirus was injected into the same subcutaneous tumour system, all mice showed significant tumour inhibition, an effect that was dependent on the indirect antitumour activities of IFN-alpha, notably a stimulation of natural killer cells. Moreover, in this case, tumour regression was observed not only for the injected subcutaneous tumours but also for the untreated tumours at distant sites. This study suggested that a local IFN-alpha gene therapy is a promising therapeutic strategy for pancreatic cancer, due to its dual mechanisms of antitumour activities and lack of significant toxicity.

Show MeSH
Related in: MedlinePlus