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Interferon-gamma Added During Bacillus Calmette-Guerin Induced Dendritic Cell Maturation Stimulates Potent Th1 Immune Responses.

Shankar G, Pestano LA, Bosch ML - J Transl Med (2003)

Bottom Line: In MLR or recall immune responses, the addition of IFN-gamma at the time of BCG-treatment did not increase the number of antigen-specific T cells but enhanced the development of IFN-gamma-producing Th1 cells.IFN-gamma co-treatment with BCG was found to induce IL-12 and, in some instances, inhibit IL-10 secretion by DC.These findings greatly enhance the potential of BCG-matured dendritic cells for use in cancer immunotherapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Northwest Biotherapeutics, Inc, 21720-23rd Dr, SE, Suite 100, Bothell, WA, U,S,A. gshanka3@cntus.jnj.com

ABSTRACT
Dendritic cells (DC) are increasingly prepared in vitro for use in immunotherapy trials. Mature DC express high levels of surface molecules needed for T cell activation and are superior at antigen-presentation than immature DC. Bacillus Calmette-Guerin (BCG) is one of several products known to induce DC maturation, and interferon (IFN)-gamma has been shown to enhance the activity of DC stimulated with certain maturation factors. In this study, we investigated the use of IFN-gamma in combination with the powerful maturation agent, BCG. The treatment of immature DC with IFN-gamma plus BCG led to the upregulation of CD54, CD80, and CD86 in comparison with BCG treatment alone. In MLR or recall immune responses, the addition of IFN-gamma at the time of BCG-treatment did not increase the number of antigen-specific T cells but enhanced the development of IFN-gamma-producing Th1 cells. In primary immune responses, on the other hand, BCG and IFN-gamma co-treated DC stimulated higher proportions of specific T cells as well as IFN-gamma secretion by these T cells. Thus the use of IFN-gamma during BCG-induced DC maturation differentially affects the nature of recall versus naïve antigen-specific T-cell responses. IFN-gamma co-treatment with BCG was found to induce IL-12 and, in some instances, inhibit IL-10 secretion by DC. These findings greatly enhance the potential of BCG-matured dendritic cells for use in cancer immunotherapy.

No MeSH data available.


IFN-γ treatment in combination with BCG induces TNF-α in a higher percentage of dendritic cells than treatment with BCG alone DC were treated with 1.6 × 106 CFU/ml of BCG, with or without 1000 U/ml IFN-γ, or left untreated (immature). Twenty hours later GolgiPlug™ was added to inhibit cytokine secretion and the cells were cultured for an additional 16 h prior to intracellular staining with anti-TNF-α-FITC and extracellular staining with anti-HLA-DR, DP, DQ-PerCP. Data shown here are from a representative experiment out of four similar experiments.
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Figure 2: IFN-γ treatment in combination with BCG induces TNF-α in a higher percentage of dendritic cells than treatment with BCG alone DC were treated with 1.6 × 106 CFU/ml of BCG, with or without 1000 U/ml IFN-γ, or left untreated (immature). Twenty hours later GolgiPlug™ was added to inhibit cytokine secretion and the cells were cultured for an additional 16 h prior to intracellular staining with anti-TNF-α-FITC and extracellular staining with anti-HLA-DR, DP, DQ-PerCP. Data shown here are from a representative experiment out of four similar experiments.

Mentions: To investigate whether this potentiating effect of IFN-γ on BCG-induced DC maturation was restricted to surface molecule expression alone, we evaluated the production of TNF-α. DC were matured for 36 h as above and the proportion of TNF-α-producing DC was determined by intracellular cytokine staining. As indicated in Figure 2, IFN-γ induced over a 3-fold increase in the proportion of BCG-matured DC producing TNF-α. Furthermore, the TNF-α levels on a per-cell-basis were noticeably higher in DC matured with the combination of BCG plus IFN-γ compared to those DC matured with BCG alone. These initial results suggested that IFN-γ enhances BCG-induced DC maturation.


Interferon-gamma Added During Bacillus Calmette-Guerin Induced Dendritic Cell Maturation Stimulates Potent Th1 Immune Responses.

Shankar G, Pestano LA, Bosch ML - J Transl Med (2003)

IFN-γ treatment in combination with BCG induces TNF-α in a higher percentage of dendritic cells than treatment with BCG alone DC were treated with 1.6 × 106 CFU/ml of BCG, with or without 1000 U/ml IFN-γ, or left untreated (immature). Twenty hours later GolgiPlug™ was added to inhibit cytokine secretion and the cells were cultured for an additional 16 h prior to intracellular staining with anti-TNF-α-FITC and extracellular staining with anti-HLA-DR, DP, DQ-PerCP. Data shown here are from a representative experiment out of four similar experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC239912&req=5

Figure 2: IFN-γ treatment in combination with BCG induces TNF-α in a higher percentage of dendritic cells than treatment with BCG alone DC were treated with 1.6 × 106 CFU/ml of BCG, with or without 1000 U/ml IFN-γ, or left untreated (immature). Twenty hours later GolgiPlug™ was added to inhibit cytokine secretion and the cells were cultured for an additional 16 h prior to intracellular staining with anti-TNF-α-FITC and extracellular staining with anti-HLA-DR, DP, DQ-PerCP. Data shown here are from a representative experiment out of four similar experiments.
Mentions: To investigate whether this potentiating effect of IFN-γ on BCG-induced DC maturation was restricted to surface molecule expression alone, we evaluated the production of TNF-α. DC were matured for 36 h as above and the proportion of TNF-α-producing DC was determined by intracellular cytokine staining. As indicated in Figure 2, IFN-γ induced over a 3-fold increase in the proportion of BCG-matured DC producing TNF-α. Furthermore, the TNF-α levels on a per-cell-basis were noticeably higher in DC matured with the combination of BCG plus IFN-γ compared to those DC matured with BCG alone. These initial results suggested that IFN-γ enhances BCG-induced DC maturation.

Bottom Line: In MLR or recall immune responses, the addition of IFN-gamma at the time of BCG-treatment did not increase the number of antigen-specific T cells but enhanced the development of IFN-gamma-producing Th1 cells.IFN-gamma co-treatment with BCG was found to induce IL-12 and, in some instances, inhibit IL-10 secretion by DC.These findings greatly enhance the potential of BCG-matured dendritic cells for use in cancer immunotherapy.

View Article: PubMed Central - HTML - PubMed

Affiliation: Northwest Biotherapeutics, Inc, 21720-23rd Dr, SE, Suite 100, Bothell, WA, U,S,A. gshanka3@cntus.jnj.com

ABSTRACT
Dendritic cells (DC) are increasingly prepared in vitro for use in immunotherapy trials. Mature DC express high levels of surface molecules needed for T cell activation and are superior at antigen-presentation than immature DC. Bacillus Calmette-Guerin (BCG) is one of several products known to induce DC maturation, and interferon (IFN)-gamma has been shown to enhance the activity of DC stimulated with certain maturation factors. In this study, we investigated the use of IFN-gamma in combination with the powerful maturation agent, BCG. The treatment of immature DC with IFN-gamma plus BCG led to the upregulation of CD54, CD80, and CD86 in comparison with BCG treatment alone. In MLR or recall immune responses, the addition of IFN-gamma at the time of BCG-treatment did not increase the number of antigen-specific T cells but enhanced the development of IFN-gamma-producing Th1 cells. In primary immune responses, on the other hand, BCG and IFN-gamma co-treated DC stimulated higher proportions of specific T cells as well as IFN-gamma secretion by these T cells. Thus the use of IFN-gamma during BCG-induced DC maturation differentially affects the nature of recall versus naïve antigen-specific T-cell responses. IFN-gamma co-treatment with BCG was found to induce IL-12 and, in some instances, inhibit IL-10 secretion by DC. These findings greatly enhance the potential of BCG-matured dendritic cells for use in cancer immunotherapy.

No MeSH data available.