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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 in  bone marrow DC (BMDC) in vitro. The primary murine DCs were infected in vitro with Adβgal or 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. For all panels, the y-axis reflects DC number and  the x-axis reflects log fluorescein intensity. The percentage of βgal-expressing DCs was determined by the right shift of the curve along the K  gate. (A) Uninfected bone marrow DCs. (B) Bone marrow DCs infected  with AdNull. (C) Bone marrow DCs infected with Adβgal.
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Figure 7: Ad vector–mediated gene transfer and expression of βgal in bone marrow DC (BMDC) in vitro. The primary murine DCs were infected in vitro with Adβgal or 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. For all panels, the y-axis reflects DC number and the x-axis reflects log fluorescein intensity. The percentage of βgal-expressing DCs was determined by the right shift of the curve along the K gate. (A) Uninfected bone marrow DCs. (B) Bone marrow DCs infected with AdNull. (C) Bone marrow DCs infected with Adβgal.

Mentions: To evaluate the ability of Ad vectors to transfer and express genes in primary bone marrow DCs, Adβgal was used to infect bone marrow DCs an moi of 100, and the cells were quantified for βgal activity at 24 h after infection by flow cytometry. Similar to the findings with XS52-Adβgal (Fig. 1, C), expression of βgal was readily detectable (95%) in bone marrow DCs infected with Adβgal (Fig. 7, C), but not in naive bone marrow DCs (A) or bone marrow DCs infected with AdNull (B), demonstrating that Ad vectors can transfer and express genes in primary DCs. Parallel studies carried out using other moi demonstrated 17% bone marrow DC cells expressing βgal with an moi of 30 and 96% at an moi of 300 (not shown). Based on these studies, all subsequent studies were carried out at an moi 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 in  bone marrow DC (BMDC) in vitro. The primary murine DCs were infected in vitro with Adβgal or 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. For all panels, the y-axis reflects DC number and  the x-axis reflects log fluorescein intensity. The percentage of βgal-expressing DCs was determined by the right shift of the curve along the K  gate. (A) Uninfected bone marrow DCs. (B) Bone marrow DCs infected  with AdNull. (C) Bone marrow DCs infected with Adβgal.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2199096&req=5

Figure 7: Ad vector–mediated gene transfer and expression of βgal in bone marrow DC (BMDC) in vitro. The primary murine DCs were infected in vitro with Adβgal or 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. For all panels, the y-axis reflects DC number and the x-axis reflects log fluorescein intensity. The percentage of βgal-expressing DCs was determined by the right shift of the curve along the K gate. (A) Uninfected bone marrow DCs. (B) Bone marrow DCs infected with AdNull. (C) Bone marrow DCs infected with Adβgal.
Mentions: To evaluate the ability of Ad vectors to transfer and express genes in primary bone marrow DCs, Adβgal was used to infect bone marrow DCs an moi of 100, and the cells were quantified for βgal activity at 24 h after infection by flow cytometry. Similar to the findings with XS52-Adβgal (Fig. 1, C), expression of βgal was readily detectable (95%) in bone marrow DCs infected with Adβgal (Fig. 7, C), but not in naive bone marrow DCs (A) or bone marrow DCs infected with AdNull (B), demonstrating that Ad vectors can transfer and express genes in primary DCs. Parallel studies carried out using other moi demonstrated 17% bone marrow DC cells expressing βgal with an moi of 30 and 96% at an moi of 300 (not shown). Based on these studies, all subsequent studies were carried out at an moi 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