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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 bone  marrow DCs infected with Adβgal. Bone marrow DCs were  transduced in vitro with AdNull  or Adβgal at moi of 100 for 2 h.  24 h later, 3 × 105 cells were administered subcutaneously into  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 βgal peptide and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells, βgal-expressing  CT26.CL25 cells, or CT26.WT pulsed with βgal peptide in vitro [CT26.WT-βgal peptide]) as well as the E22 βgal-expressing allogeneic tumor cell  line. (A) CTL response in mice immunized with bone marrow DCs infected with AdNull. (B) CTL response in mice immunized with bone marrow  DCs infected with Adβgal.
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Figure 8: Induction of CTL response in BALB/c mice after in vivo administration of bone marrow DCs infected with Adβgal. Bone marrow DCs were transduced in vitro with AdNull or Adβgal at moi of 100 for 2 h. 24 h later, 3 × 105 cells were administered subcutaneously into 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 βgal peptide and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells, βgal-expressing CT26.CL25 cells, or CT26.WT pulsed with βgal peptide in vitro [CT26.WT-βgal peptide]) as well as the E22 βgal-expressing allogeneic tumor cell line. (A) CTL response in mice immunized with bone marrow DCs infected with AdNull. (B) CTL response in mice immunized with bone marrow DCs infected with Adβgal.

Mentions: To evaluate if genetically modified primary bone marrow DCs would induce an antigen-specific CTL response in vivo, splenocytes from mice immunized with subcutaneous administration of bone marrow DCs transduced with Adβgal were stimulated in vitro with βgal peptide and were then evaluated for specific cell lysis against a panel of target cells in a standard 51Cr–release assay. Immunization with bone marrow DC–Adβgal cells induced a βgal-specific CTL response (Fig. 8, B), whereas the negative control bone marrow DC–AdNull cells did not (A). Only negligible lysis of non–βgal-expressing CT26.WT target cells were detected in mice immunized with bone marrow DC–Adβgal cells, indicating that the cellular immune response induced was βgal specific. This specific cytotoxicity was mediated predominately by MHC-restricted CTLs rather than by natural killer cells since E22, an MHC-mismatched (H-2b) βgal-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 bone  marrow DCs infected with Adβgal. Bone marrow DCs were  transduced in vitro with AdNull  or Adβgal at moi of 100 for 2 h.  24 h later, 3 × 105 cells were administered subcutaneously into  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 βgal peptide and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells, βgal-expressing  CT26.CL25 cells, or CT26.WT pulsed with βgal peptide in vitro [CT26.WT-βgal peptide]) as well as the E22 βgal-expressing allogeneic tumor cell  line. (A) CTL response in mice immunized with bone marrow DCs infected with AdNull. (B) CTL response in mice immunized with bone marrow  DCs infected with Adβgal.
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Related In: Results  -  Collection

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Figure 8: Induction of CTL response in BALB/c mice after in vivo administration of bone marrow DCs infected with Adβgal. Bone marrow DCs were transduced in vitro with AdNull or Adβgal at moi of 100 for 2 h. 24 h later, 3 × 105 cells were administered subcutaneously into 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 βgal peptide and then assayed for specific cell lysis against three syngeneic colon carcinoma target cells (parental CT26.WT cells, βgal-expressing CT26.CL25 cells, or CT26.WT pulsed with βgal peptide in vitro [CT26.WT-βgal peptide]) as well as the E22 βgal-expressing allogeneic tumor cell line. (A) CTL response in mice immunized with bone marrow DCs infected with AdNull. (B) CTL response in mice immunized with bone marrow DCs infected with Adβgal.
Mentions: To evaluate if genetically modified primary bone marrow DCs would induce an antigen-specific CTL response in vivo, splenocytes from mice immunized with subcutaneous administration of bone marrow DCs transduced with Adβgal were stimulated in vitro with βgal peptide and were then evaluated for specific cell lysis against a panel of target cells in a standard 51Cr–release assay. Immunization with bone marrow DC–Adβgal cells induced a βgal-specific CTL response (Fig. 8, B), whereas the negative control bone marrow DC–AdNull cells did not (A). Only negligible lysis of non–βgal-expressing CT26.WT target cells were detected in mice immunized with bone marrow DC–Adβgal cells, indicating that the cellular immune response induced was βgal specific. This specific cytotoxicity was mediated predominately by MHC-restricted CTLs rather than by natural killer cells since E22, an MHC-mismatched (H-2b) βgal-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