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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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Induction of CTL  response in BALB/c mice after  in vivo administration of Ad╬▓gal-infected XS52 DCs. The  XS52 DCs were transduced in  vitro with AdNull or Ad╬▓gal at  moi of 100 for 2 h. 24 h later, 3 ├Ś  105 cells were administered subcutaneously to syngeneic BALB/ c mice. 14 d later, splenocytes  harvested from immunized mice  were stimulated for 5 d in vitro  with syngeneic fibroblasts  (SVBalb) pulsed with a ╬▓gal peptide, and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells,  ╬▓gal-expressing CT26.CL25 cells, or CT26.WT pulsed with ╬▓gal peptide in vitro [CT26.WT-╬▓gal peptide]) as well as the E22 ╬▓gal-expressing allogeneic tumor cell line. (A) CTL response in mice immunized with XS52 DCs infected with AdNull. (B) CTL response in mice immunized with XS52  DCs infected with Ad╬▓gal.
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Figure 2: Induction of CTL response in BALB/c mice after in vivo administration of Ad╬▓gal-infected XS52 DCs. The XS52 DCs were transduced in vitro with AdNull or Ad╬▓gal at moi of 100 for 2 h. 24 h later, 3 ├Ś 105 cells were administered subcutaneously to syngeneic BALB/ c mice. 14 d later, splenocytes harvested from immunized mice were stimulated for 5 d in vitro with syngeneic fibroblasts (SVBalb) pulsed with a ╬▓gal peptide, and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells, ╬▓gal-expressing CT26.CL25 cells, or CT26.WT pulsed with ╬▓gal peptide in vitro [CT26.WT-╬▓gal peptide]) as well as the E22 ╬▓gal-expressing allogeneic tumor cell line. (A) CTL response in mice immunized with XS52 DCs infected with AdNull. (B) CTL response in mice immunized with XS52 DCs infected with Ad╬▓gal.

Mentions: To determine if Ad-mediated gene expression in DCs would induce an antigen-specific CTL response in vivo, BALB/c mice were first immunized with 3 ├Ś 105 XS52 cells that had been modified in vitro with AdNull- or Ad╬▓gal-infection. Splenocytes from immunized mice were then harvested and stimulated in vitro with ╬▓gal peptide and were then assayed for specific cell lysis against a panel of target cells in a standared 51CrÔÇôrelease assay. Although immunization of naive XS52 or XS52-AdNull infected cells did not result in a measurable ╬▓gal-specific CTL response (Fig. 2, A), significant lysis of ╬▓gal-expressing CT26.CL25 target cells or ╬▓gal peptide-pulsed CT26.WT target cells were observed in mice immunized with XS52-Ad╬▓gal (B). Only negligible lysis of nonÔÇô╬▓gal-expressing CT26.WT target cells was detected in mice immunized with XS52-Ad╬▓gal, indicating that the cellular immune response induced by XS52-Ad╬▓gal was ╬▓gal specific. This specific cytotoxicity was mediated predominantly by MHC-restricted CTLs rather than by natural killer cells since E22, an MHC-mismatched (H-2b), ╬▓gal-expressing murine tumor cell line, was not significantly lysed in the same assay.


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)

Induction of CTL  response in BALB/c mice after  in vivo administration of Ad╬▓gal-infected XS52 DCs. The  XS52 DCs were transduced in  vitro with AdNull or Ad╬▓gal at  moi of 100 for 2 h. 24 h later, 3 ├Ś  105 cells were administered subcutaneously to syngeneic BALB/ c mice. 14 d later, splenocytes  harvested from immunized mice  were stimulated for 5 d in vitro  with syngeneic fibroblasts  (SVBalb) pulsed with a ╬▓gal peptide, and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells,  ╬▓gal-expressing CT26.CL25 cells, or CT26.WT pulsed with ╬▓gal peptide in vitro [CT26.WT-╬▓gal peptide]) as well as the E22 ╬▓gal-expressing allogeneic tumor cell line. (A) CTL response in mice immunized with XS52 DCs infected with AdNull. (B) CTL response in mice immunized with XS52  DCs infected with Ad╬▓gal.
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Related In: Results  -  Collection

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Figure 2: Induction of CTL response in BALB/c mice after in vivo administration of Ad╬▓gal-infected XS52 DCs. The XS52 DCs were transduced in vitro with AdNull or Ad╬▓gal at moi of 100 for 2 h. 24 h later, 3 ├Ś 105 cells were administered subcutaneously to syngeneic BALB/ c mice. 14 d later, splenocytes harvested from immunized mice were stimulated for 5 d in vitro with syngeneic fibroblasts (SVBalb) pulsed with a ╬▓gal peptide, and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells, ╬▓gal-expressing CT26.CL25 cells, or CT26.WT pulsed with ╬▓gal peptide in vitro [CT26.WT-╬▓gal peptide]) as well as the E22 ╬▓gal-expressing allogeneic tumor cell line. (A) CTL response in mice immunized with XS52 DCs infected with AdNull. (B) CTL response in mice immunized with XS52 DCs infected with Ad╬▓gal.
Mentions: To determine if Ad-mediated gene expression in DCs would induce an antigen-specific CTL response in vivo, BALB/c mice were first immunized with 3 ├Ś 105 XS52 cells that had been modified in vitro with AdNull- or Ad╬▓gal-infection. Splenocytes from immunized mice were then harvested and stimulated in vitro with ╬▓gal peptide and were then assayed for specific cell lysis against a panel of target cells in a standared 51CrÔÇôrelease assay. Although immunization of naive XS52 or XS52-AdNull infected cells did not result in a measurable ╬▓gal-specific CTL response (Fig. 2, A), significant lysis of ╬▓gal-expressing CT26.CL25 target cells or ╬▓gal peptide-pulsed CT26.WT target cells were observed in mice immunized with XS52-Ad╬▓gal (B). Only negligible lysis of nonÔÇô╬▓gal-expressing CT26.WT target cells was detected in mice immunized with XS52-Ad╬▓gal, indicating that the cellular immune response induced by XS52-Ad╬▓gal was ╬▓gal specific. This specific cytotoxicity was mediated predominantly by MHC-restricted CTLs rather than by natural killer cells since E22, an MHC-mismatched (H-2b), ╬▓gal-expressing murine tumor cell line, was not significantly lysed in the same assay.

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