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Dendritic cells retrovirally transduced with a model antigen gene are therapeutically effective against established pulmonary metastases.

Specht JM, Wang G, Do MT, Lam JS, Royal RE, Reeves ME, Rosenberg SA, Hwu P - J. Exp. Med. (1997)

Bottom Line: DC pulsed with tumor-associated antigen (TAA) peptide or protein have recently been demonstrated to elicit antigen-specific protective antitumor immunity in a number of murine models.In addition, coculture of beta-gal transduced DC with a beta-gal-specific T cell line (CTLx) resulted in the production of large amounts of interferon-gamma, demonstrating that transduced DCs could process and present endogenously expressed beta-gal.Treatment with beta-gal-transduced DC significantly reduced the number of pulmonary metastatic nodules compared with treatment with Hank's balanced salt solution or DCs transduced with rat HER-2/neu.

View Article: PubMed Central - PubMed

Affiliation: Surgery Branch, National Cancer Institute, Bethesda, Maryland 20892, USA.

ABSTRACT
Dendritic cells (DCs) are bone marrow-derived leukocytes that function as potent antigen presenting cells capable of initiating T cell-dependent responses from quiescent lymphocytes. DC pulsed with tumor-associated antigen (TAA) peptide or protein have recently been demonstrated to elicit antigen-specific protective antitumor immunity in a number of murine models. Transduction of DCs with TAA genes may allow stable, prolonged antigen expression as well as the potential for presentation of multiple, or unidentified, epitopes in association with major histocompatibility complex class I and/or class II molecules. To evaluate the potential efficacy of retrovirally transduced DCs, bone marrow cells harvested from BALB/c mice were transduced with either a model antigen gene encoding beta-galactosidase (beta-gal) or a control gene encoding rat HER-2/neu (Neu) by coculture with irradiated ecotropic retroviral producer lines. Bone marrow cells were differentiated into DC in vitro using granulocyte/macrophage colony-stimulating factor and interleukin-4. After 7 d in culture, cells were 45-78% double positive for DC phenotypic cell surface markers by FACS(R) analysis, and DC transduced with beta-gal were 41-72% positive for beta-gal expression by X-gal staining. In addition, coculture of beta-gal transduced DC with a beta-gal-specific T cell line (CTLx) resulted in the production of large amounts of interferon-gamma, demonstrating that transduced DCs could process and present endogenously expressed beta-gal. DC transduced with beta-gal and control rat HER-2/neu were then used to treat 3-d lung metastases in mice bearing an experimental murine tumor CT26.CL25, expressing the model antigen, beta-gal. Treatment with beta-gal-transduced DC significantly reduced the number of pulmonary metastatic nodules compared with treatment with Hank's balanced salt solution or DCs transduced with rat HER-2/neu. In addition, immunization with beta-gal-transduced DCs resulted in the generation of antigen-specific cytotoxic T lymphocytes (CTLs), which were significantly more reactive against relevant tumor targets than CTLs generated from mice immunized with DCs pulsed with the Ld-restricted beta-gal peptide. The results observed in this rapidly lethal tumor model suggest that DCs transduced with TAA may be a useful treatment modality in tumor immunotherapy.

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Active immunotherapy with retrovirally transduced or peptide-pulsed DCs significantly reduces the number of established pulmonary metastases in this 3-d tumor model. (A) 8–10 BALB/c mice/group  were injected intravenously with 3 × 105 CT26.CL25 tumor cells. On  days 3 and 6 after tumor challenge, mice received intravenous injections  of 4 × 105 transduced or peptide-pulsed DCs, fresh peptide-pulsed splenocytes, or HBSS alone. On day 12 after tumor challenge, lungs were  harvested and pulmonary nodules were enumerated in a blinded fashion.  (B) BALB/c mice were challenged with 3 × 105 CT26.CL25 tumor cells.  On days 3 and 6 after tumor challenge, mice received intravenous injections of transduced DCs or fresh peptide-pulsed splenocytes at varying  doses (4–16 × 105). On day 12 after tumor challenge, lungs were harvested and pulmonary nodules were enumerated in a blinded fashion.
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Figure 4: Active immunotherapy with retrovirally transduced or peptide-pulsed DCs significantly reduces the number of established pulmonary metastases in this 3-d tumor model. (A) 8–10 BALB/c mice/group were injected intravenously with 3 × 105 CT26.CL25 tumor cells. On days 3 and 6 after tumor challenge, mice received intravenous injections of 4 × 105 transduced or peptide-pulsed DCs, fresh peptide-pulsed splenocytes, or HBSS alone. On day 12 after tumor challenge, lungs were harvested and pulmonary nodules were enumerated in a blinded fashion. (B) BALB/c mice were challenged with 3 × 105 CT26.CL25 tumor cells. On days 3 and 6 after tumor challenge, mice received intravenous injections of transduced DCs or fresh peptide-pulsed splenocytes at varying doses (4–16 × 105). On day 12 after tumor challenge, lungs were harvested and pulmonary nodules were enumerated in a blinded fashion.

Mentions: To determine the therapeutic efficacy of retrovirally transduced and peptide-pulsed DCs against established pulmonary metastases, BALB/c mice were challenged with 3 × 105 CT26.CL25 tumor cells intravenously and treated intravenously with either 4 × 105 β-gal or HER-2/neu–transduced DCs, 4 × 105 peptide-pulsed DCs, 4 × 105 βgP-pulsed splenocytes, or HBSS on days 3 and 6 after tumor challenge. On day 12, the number of pulmonary metastastic nodules were enumerated in a blinded fashion. Mice treated with DCs transduced with β-gal (DC–β-gal) or pulsed with βgP (DC– βgP) showed a significant reduction in the number of pulmonary metastases compared with mice treated with HER-2/neu–transduced (DC–Neu) or P1A peptide-pulsed DCs (DC–P1A) (Fig. 4 A). The unique role of DCs in this response is evidenced by the lack of treatment effect seen in mice treated with βgP-pulsed splenocytes (splen– βgP). These results are representative of data obtained from four independent experiments.


Dendritic cells retrovirally transduced with a model antigen gene are therapeutically effective against established pulmonary metastases.

Specht JM, Wang G, Do MT, Lam JS, Royal RE, Reeves ME, Rosenberg SA, Hwu P - J. Exp. Med. (1997)

Active immunotherapy with retrovirally transduced or peptide-pulsed DCs significantly reduces the number of established pulmonary metastases in this 3-d tumor model. (A) 8–10 BALB/c mice/group  were injected intravenously with 3 × 105 CT26.CL25 tumor cells. On  days 3 and 6 after tumor challenge, mice received intravenous injections  of 4 × 105 transduced or peptide-pulsed DCs, fresh peptide-pulsed splenocytes, or HBSS alone. On day 12 after tumor challenge, lungs were  harvested and pulmonary nodules were enumerated in a blinded fashion.  (B) BALB/c mice were challenged with 3 × 105 CT26.CL25 tumor cells.  On days 3 and 6 after tumor challenge, mice received intravenous injections of transduced DCs or fresh peptide-pulsed splenocytes at varying  doses (4–16 × 105). On day 12 after tumor challenge, lungs were harvested and pulmonary nodules were enumerated in a blinded fashion.
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Related In: Results  -  Collection

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Figure 4: Active immunotherapy with retrovirally transduced or peptide-pulsed DCs significantly reduces the number of established pulmonary metastases in this 3-d tumor model. (A) 8–10 BALB/c mice/group were injected intravenously with 3 × 105 CT26.CL25 tumor cells. On days 3 and 6 after tumor challenge, mice received intravenous injections of 4 × 105 transduced or peptide-pulsed DCs, fresh peptide-pulsed splenocytes, or HBSS alone. On day 12 after tumor challenge, lungs were harvested and pulmonary nodules were enumerated in a blinded fashion. (B) BALB/c mice were challenged with 3 × 105 CT26.CL25 tumor cells. On days 3 and 6 after tumor challenge, mice received intravenous injections of transduced DCs or fresh peptide-pulsed splenocytes at varying doses (4–16 × 105). On day 12 after tumor challenge, lungs were harvested and pulmonary nodules were enumerated in a blinded fashion.
Mentions: To determine the therapeutic efficacy of retrovirally transduced and peptide-pulsed DCs against established pulmonary metastases, BALB/c mice were challenged with 3 × 105 CT26.CL25 tumor cells intravenously and treated intravenously with either 4 × 105 β-gal or HER-2/neu–transduced DCs, 4 × 105 peptide-pulsed DCs, 4 × 105 βgP-pulsed splenocytes, or HBSS on days 3 and 6 after tumor challenge. On day 12, the number of pulmonary metastastic nodules were enumerated in a blinded fashion. Mice treated with DCs transduced with β-gal (DC–β-gal) or pulsed with βgP (DC– βgP) showed a significant reduction in the number of pulmonary metastases compared with mice treated with HER-2/neu–transduced (DC–Neu) or P1A peptide-pulsed DCs (DC–P1A) (Fig. 4 A). The unique role of DCs in this response is evidenced by the lack of treatment effect seen in mice treated with βgP-pulsed splenocytes (splen– βgP). These results are representative of data obtained from four independent experiments.

Bottom Line: DC pulsed with tumor-associated antigen (TAA) peptide or protein have recently been demonstrated to elicit antigen-specific protective antitumor immunity in a number of murine models.In addition, coculture of beta-gal transduced DC with a beta-gal-specific T cell line (CTLx) resulted in the production of large amounts of interferon-gamma, demonstrating that transduced DCs could process and present endogenously expressed beta-gal.Treatment with beta-gal-transduced DC significantly reduced the number of pulmonary metastatic nodules compared with treatment with Hank's balanced salt solution or DCs transduced with rat HER-2/neu.

View Article: PubMed Central - PubMed

Affiliation: Surgery Branch, National Cancer Institute, Bethesda, Maryland 20892, USA.

ABSTRACT
Dendritic cells (DCs) are bone marrow-derived leukocytes that function as potent antigen presenting cells capable of initiating T cell-dependent responses from quiescent lymphocytes. DC pulsed with tumor-associated antigen (TAA) peptide or protein have recently been demonstrated to elicit antigen-specific protective antitumor immunity in a number of murine models. Transduction of DCs with TAA genes may allow stable, prolonged antigen expression as well as the potential for presentation of multiple, or unidentified, epitopes in association with major histocompatibility complex class I and/or class II molecules. To evaluate the potential efficacy of retrovirally transduced DCs, bone marrow cells harvested from BALB/c mice were transduced with either a model antigen gene encoding beta-galactosidase (beta-gal) or a control gene encoding rat HER-2/neu (Neu) by coculture with irradiated ecotropic retroviral producer lines. Bone marrow cells were differentiated into DC in vitro using granulocyte/macrophage colony-stimulating factor and interleukin-4. After 7 d in culture, cells were 45-78% double positive for DC phenotypic cell surface markers by FACS(R) analysis, and DC transduced with beta-gal were 41-72% positive for beta-gal expression by X-gal staining. In addition, coculture of beta-gal transduced DC with a beta-gal-specific T cell line (CTLx) resulted in the production of large amounts of interferon-gamma, demonstrating that transduced DCs could process and present endogenously expressed beta-gal. DC transduced with beta-gal and control rat HER-2/neu were then used to treat 3-d lung metastases in mice bearing an experimental murine tumor CT26.CL25, expressing the model antigen, beta-gal. Treatment with beta-gal-transduced DC significantly reduced the number of pulmonary metastatic nodules compared with treatment with Hank's balanced salt solution or DCs transduced with rat HER-2/neu. In addition, immunization with beta-gal-transduced DCs resulted in the generation of antigen-specific cytotoxic T lymphocytes (CTLs), which were significantly more reactive against relevant tumor targets than CTLs generated from mice immunized with DCs pulsed with the Ld-restricted beta-gal peptide. The results observed in this rapidly lethal tumor model suggest that DCs transduced with TAA may be a useful treatment modality in tumor immunotherapy.

Show MeSH
Related in: MedlinePlus