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Dendritic cells genetically modified with an adenovirus vector encoding the cDNA for a model antigen induce protective and therapeutic antitumor immunity.

Song W, Kong HL, Carpenter H, Torii H, Granstein R, Rafii S, Moore MA, Crystal RG - J. Exp. Med. (1997)

Bottom Line: Dendritic cells (DCs) are potent antigen-presenting cells that play a critical role in the initiation of antitumor immune responses.In this study, we show that genetic modifications of a murine epidermis-derived DC line and primary bone marrow-derived DCs to express a model antigen beta-galactosidase (betagal) can be achieved through the use of a replication-deficient, recombinant adenovirus vector, and that the modified DCs are capable of eliciting antigen-specific, MHC-restricted CTL responses.Importantly, using a murine metastatic lung tumor model with syngeneic colon carcinoma cells expressing betagal, we show that immunization of mice with the genetically modified DC line or bone marrow DCs confers potent protection against a lethal tumor challenge, as well as suppression of preestablished tumors, resulting in a significant survival advantage.

View Article: PubMed Central - PubMed

Affiliation: Division of Pulmonary and Critical Care Medicine, The New York Hospital-Cornell Medical Center 10021, USA.

ABSTRACT
Dendritic cells (DCs) are potent antigen-presenting cells that play a critical role in the initiation of antitumor immune responses. In this study, we show that genetic modifications of a murine epidermis-derived DC line and primary bone marrow-derived DCs to express a model antigen beta-galactosidase (betagal) can be achieved through the use of a replication-deficient, recombinant adenovirus vector, and that the modified DCs are capable of eliciting antigen-specific, MHC-restricted CTL responses. Importantly, using a murine metastatic lung tumor model with syngeneic colon carcinoma cells expressing betagal, we show that immunization of mice with the genetically modified DC line or bone marrow DCs confers potent protection against a lethal tumor challenge, as well as suppression of preestablished tumors, resulting in a significant survival advantage. We conclude that genetic modification of DCs to express antigens that are also expressed in tumors can lead to antigen-specific, antitumor killer cells, with a concomitant resistance to tumor challenge and a decrease in the size of existing tumors.

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Ad vector–mediated gene transfer and expression of βgal to  the murine XS52 DC line in vitro. The XS52 cells were infected in vitro  with an Ad vector expressing βgal (Adβgal) or the AdNull control vector  at moi of 100 for 2 h. 24 h later, βgal expression was quantified by flow  cytometry using fluorescein di-β-galactoside. Cells were stained with  propidum iodide to facilitate discrimination among live and dead cells.  For all panels, the y-axis reflects DC number and the x-axis reflects log  fluorescein intensity. The K gate has been set according to the negative  control in A. The percentage of βgal-expressing DCs was determined by  the right shift of the curve along the K gate. (A) Uninfected XS52 cells.  (B) XS52 infected with AdNull. (C) XS52 infected with Adβgal.
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Figure 1: Ad vector–mediated gene transfer and expression of βgal to the murine XS52 DC line in vitro. The XS52 cells were infected in vitro with an Ad vector expressing βgal (Adβgal) or the AdNull control vector at moi of 100 for 2 h. 24 h later, βgal expression was quantified by flow cytometry using fluorescein di-β-galactoside. Cells were stained with propidum iodide to facilitate discrimination among live and dead cells. For all panels, the y-axis reflects DC number and the x-axis reflects log fluorescein intensity. The K gate has been set according to the negative control in A. The percentage of βgal-expressing DCs was determined by the right shift of the curve along the K gate. (A) Uninfected XS52 cells. (B) XS52 infected with AdNull. (C) XS52 infected with Adβgal.

Mentions: To evaluate the ability of Ad vectors to transfer and express genes in XS52 DCs, XS52 DCs were infected with Adβgal, and the cells were quantified for βgal activity at 24 h after infection by flow cytometry. Although naive XS52 (Fig. 1, A) and AdNull-infected XS52 (B) showed only background levels of βgal activity, expression of βgal was readily detected in the majority (73%) of XS52 infected with Adβgal at moi of 100 (C), demonstrating that successful gene transfer and expression can be achieved in DCs using an Ad vector. Parallel studies carried out using other moi demonstrated 14% XS52 cells expressing βgal with a moi of 30 and 62% at a moi of 300 (not shown). On the basis of these studies, all subsequent studies were carried out with a moi of 100.


Dendritic cells genetically modified with an adenovirus vector encoding the cDNA for a model antigen induce protective and therapeutic antitumor immunity.

Song W, Kong HL, Carpenter H, Torii H, Granstein R, Rafii S, Moore MA, Crystal RG - J. Exp. Med. (1997)

Ad vector–mediated gene transfer and expression of βgal to  the murine XS52 DC line in vitro. The XS52 cells were infected in vitro  with an Ad vector expressing βgal (Adβgal) or the AdNull control vector  at moi of 100 for 2 h. 24 h later, βgal expression was quantified by flow  cytometry using fluorescein di-β-galactoside. Cells were stained with  propidum iodide to facilitate discrimination among live and dead cells.  For all panels, the y-axis reflects DC number and the x-axis reflects log  fluorescein intensity. The K gate has been set according to the negative  control in A. The percentage of βgal-expressing DCs was determined by  the right shift of the curve along the K gate. (A) Uninfected XS52 cells.  (B) XS52 infected with AdNull. (C) XS52 infected with Adβgal.
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Related In: Results  -  Collection

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Figure 1: Ad vector–mediated gene transfer and expression of βgal to the murine XS52 DC line in vitro. The XS52 cells were infected in vitro with an Ad vector expressing βgal (Adβgal) or the AdNull control vector at moi of 100 for 2 h. 24 h later, βgal expression was quantified by flow cytometry using fluorescein di-β-galactoside. Cells were stained with propidum iodide to facilitate discrimination among live and dead cells. For all panels, the y-axis reflects DC number and the x-axis reflects log fluorescein intensity. The K gate has been set according to the negative control in A. The percentage of βgal-expressing DCs was determined by the right shift of the curve along the K gate. (A) Uninfected XS52 cells. (B) XS52 infected with AdNull. (C) XS52 infected with Adβgal.
Mentions: To evaluate the ability of Ad vectors to transfer and express genes in XS52 DCs, XS52 DCs were infected with Adβgal, and the cells were quantified for βgal activity at 24 h after infection by flow cytometry. Although naive XS52 (Fig. 1, A) and AdNull-infected XS52 (B) showed only background levels of βgal activity, expression of βgal was readily detected in the majority (73%) of XS52 infected with Adβgal at moi of 100 (C), demonstrating that successful gene transfer and expression can be achieved in DCs using an Ad vector. Parallel studies carried out using other moi demonstrated 14% XS52 cells expressing βgal with a moi of 30 and 62% at a moi of 300 (not shown). On the basis of these studies, all subsequent studies were carried out with a moi of 100.

Bottom Line: Dendritic cells (DCs) are potent antigen-presenting cells that play a critical role in the initiation of antitumor immune responses.In this study, we show that genetic modifications of a murine epidermis-derived DC line and primary bone marrow-derived DCs to express a model antigen beta-galactosidase (betagal) can be achieved through the use of a replication-deficient, recombinant adenovirus vector, and that the modified DCs are capable of eliciting antigen-specific, MHC-restricted CTL responses.Importantly, using a murine metastatic lung tumor model with syngeneic colon carcinoma cells expressing betagal, we show that immunization of mice with the genetically modified DC line or bone marrow DCs confers potent protection against a lethal tumor challenge, as well as suppression of preestablished tumors, resulting in a significant survival advantage.

View Article: PubMed Central - PubMed

Affiliation: Division of Pulmonary and Critical Care Medicine, The New York Hospital-Cornell Medical Center 10021, USA.

ABSTRACT
Dendritic cells (DCs) are potent antigen-presenting cells that play a critical role in the initiation of antitumor immune responses. In this study, we show that genetic modifications of a murine epidermis-derived DC line and primary bone marrow-derived DCs to express a model antigen beta-galactosidase (betagal) can be achieved through the use of a replication-deficient, recombinant adenovirus vector, and that the modified DCs are capable of eliciting antigen-specific, MHC-restricted CTL responses. Importantly, using a murine metastatic lung tumor model with syngeneic colon carcinoma cells expressing betagal, we show that immunization of mice with the genetically modified DC line or bone marrow DCs confers potent protection against a lethal tumor challenge, as well as suppression of preestablished tumors, resulting in a significant survival advantage. We conclude that genetic modification of DCs to express antigens that are also expressed in tumors can lead to antigen-specific, antitumor killer cells, with a concomitant resistance to tumor challenge and a decrease in the size of existing tumors.

Show MeSH
Related in: MedlinePlus