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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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Protection against lethal tumor challenge using modified XS52 DCs. Shown are examples and  quantitative data of protection against lung metastases after XS52 immunization, using the βgal as a model antigen, BALB/c mice, and syngeneic CT26.CL25 colon carcinoma cells (expressing βgal). XS52 cells were genetically modified by in vitro infection with AdNull or Adβgal (moi 100, 2 h). 24 h after infection, 3 × 105  XS52-AdNull or XS52-Adβgal were injected subcutaneously to syngeneic BALB/c mice. 14 d later, the mice  were challenged with intravenous administration of 3 × 105 CT26.CL25 cells. 12–16 d after tumor challenge,  the mice were killed, their lungs harvested, fixed, and stained for βgal expression with X-Gal. (A) Example of  lungs from a nonimmunized mouse. (B) Example of lungs from a mouse immunized with XS52-AdNull. (C  and D) Examples of lungs from mice immunized with XS52-Adβgal. (E) Quantification of the number of lung  metastases in nonimmunized, XS52-AdNull–immunized and XS52-Adβgal–immunized mice after tumor challenge. Using a dissecting microscope, surface blue-staining (βgal+) lung metastases were enumerated. Each data  point represents an individual animal. Only metastatic deposits ⩽250 could be reliably enumerated; lungs with  >250 metastases were assigned an empirical number of 250.
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Figure 3: Protection against lethal tumor challenge using modified XS52 DCs. Shown are examples and quantitative data of protection against lung metastases after XS52 immunization, using the βgal as a model antigen, BALB/c mice, and syngeneic CT26.CL25 colon carcinoma cells (expressing βgal). XS52 cells were genetically modified by in vitro infection with AdNull or Adβgal (moi 100, 2 h). 24 h after infection, 3 × 105 XS52-AdNull or XS52-Adβgal were injected subcutaneously to syngeneic BALB/c mice. 14 d later, the mice were challenged with intravenous administration of 3 × 105 CT26.CL25 cells. 12–16 d after tumor challenge, the mice were killed, their lungs harvested, fixed, and stained for βgal expression with X-Gal. (A) Example of lungs from a nonimmunized mouse. (B) Example of lungs from a mouse immunized with XS52-AdNull. (C and D) Examples of lungs from mice immunized with XS52-Adβgal. (E) Quantification of the number of lung metastases in nonimmunized, XS52-AdNull–immunized and XS52-Adβgal–immunized mice after tumor challenge. Using a dissecting microscope, surface blue-staining (βgal+) lung metastases were enumerated. Each data point represents an individual animal. Only metastatic deposits ⩽250 could be reliably enumerated; lungs with >250 metastases were assigned an empirical number of 250.

Mentions: Between 12 and 16 d after an intravenous tumor challenge, tumor-bearing mice which were not treated showed signs of respiratory distress and developed hemorrhagic pleural effusion and diffuse lung metastases (Fig. 3, A). Similarly, mice which were previously immunized with XS52-AdNull developed numerous lung metastases (B). In marked contrast, mice which were previously immunized with XS52-Adβgal showed very few visible lung metastases under the dissecting microscope (C and D), indicating that immunization of mice with DCs genetically modified to express the βgal model antigen could effectively protect against a lethal tumor challenge. Quantitative analysis of the number of lung metastases confirmed the significant difference between XS52-Adβgal–immunized mice and untreated mice (P <0.0001) or XS52-AdNull–immunized mice (E). Further, immunization with XS52-Adβgal that had been frozen and thawed did not confer protection when compared to untreated and XS52-AdNull–immunized groups (P >0.1; not shown), i.e., live DCs were a prerequisite to generate an effective antitumor immunity.


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)

Protection against lethal tumor challenge using modified XS52 DCs. Shown are examples and  quantitative data of protection against lung metastases after XS52 immunization, using the βgal as a model antigen, BALB/c mice, and syngeneic CT26.CL25 colon carcinoma cells (expressing βgal). XS52 cells were genetically modified by in vitro infection with AdNull or Adβgal (moi 100, 2 h). 24 h after infection, 3 × 105  XS52-AdNull or XS52-Adβgal were injected subcutaneously to syngeneic BALB/c mice. 14 d later, the mice  were challenged with intravenous administration of 3 × 105 CT26.CL25 cells. 12–16 d after tumor challenge,  the mice were killed, their lungs harvested, fixed, and stained for βgal expression with X-Gal. (A) Example of  lungs from a nonimmunized mouse. (B) Example of lungs from a mouse immunized with XS52-AdNull. (C  and D) Examples of lungs from mice immunized with XS52-Adβgal. (E) Quantification of the number of lung  metastases in nonimmunized, XS52-AdNull–immunized and XS52-Adβgal–immunized mice after tumor challenge. Using a dissecting microscope, surface blue-staining (βgal+) lung metastases were enumerated. Each data  point represents an individual animal. Only metastatic deposits ⩽250 could be reliably enumerated; lungs with  >250 metastases were assigned an empirical number of 250.
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Related In: Results  -  Collection

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Figure 3: Protection against lethal tumor challenge using modified XS52 DCs. Shown are examples and quantitative data of protection against lung metastases after XS52 immunization, using the βgal as a model antigen, BALB/c mice, and syngeneic CT26.CL25 colon carcinoma cells (expressing βgal). XS52 cells were genetically modified by in vitro infection with AdNull or Adβgal (moi 100, 2 h). 24 h after infection, 3 × 105 XS52-AdNull or XS52-Adβgal were injected subcutaneously to syngeneic BALB/c mice. 14 d later, the mice were challenged with intravenous administration of 3 × 105 CT26.CL25 cells. 12–16 d after tumor challenge, the mice were killed, their lungs harvested, fixed, and stained for βgal expression with X-Gal. (A) Example of lungs from a nonimmunized mouse. (B) Example of lungs from a mouse immunized with XS52-AdNull. (C and D) Examples of lungs from mice immunized with XS52-Adβgal. (E) Quantification of the number of lung metastases in nonimmunized, XS52-AdNull–immunized and XS52-Adβgal–immunized mice after tumor challenge. Using a dissecting microscope, surface blue-staining (βgal+) lung metastases were enumerated. Each data point represents an individual animal. Only metastatic deposits ⩽250 could be reliably enumerated; lungs with >250 metastases were assigned an empirical number of 250.
Mentions: Between 12 and 16 d after an intravenous tumor challenge, tumor-bearing mice which were not treated showed signs of respiratory distress and developed hemorrhagic pleural effusion and diffuse lung metastases (Fig. 3, A). Similarly, mice which were previously immunized with XS52-AdNull developed numerous lung metastases (B). In marked contrast, mice which were previously immunized with XS52-Adβgal showed very few visible lung metastases under the dissecting microscope (C and D), indicating that immunization of mice with DCs genetically modified to express the βgal model antigen could effectively protect against a lethal tumor challenge. Quantitative analysis of the number of lung metastases confirmed the significant difference between XS52-Adβgal–immunized mice and untreated mice (P <0.0001) or XS52-AdNull–immunized mice (E). Further, immunization with XS52-Adβgal that had been frozen and thawed did not confer protection when compared to untreated and XS52-AdNull–immunized groups (P >0.1; not shown), i.e., live DCs were a prerequisite to generate an effective antitumor immunity.

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