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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 affects the quality but not quantity of the allogeneic T cell MLR response stimulated by BCG matured DC DC were treated with 4.1 × 105 CFU/ml of BCG alone, BCG plus 1000 U/ml IFN-γ or left untreated (immature) for 24 h prior to mixed leukocyte culture. Varying numbers of DC were added to 2 × 105 enriched T cells in triplicate wells of a 96-well microculture plate and cultured for 5 days. The cells were pulsed with 3H-thymidine for an additional 24 h before harvesting and quantitation of incorporated radioactivity in a scintillation counter (A). Immediately preceding the 3H-thymidine pulse, culture supernatants were collected for evaluating IFN-γ secretion (B). Results shown in each panel are from a representative experiment out of three similar experiments.
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Figure 3: IFN-γ treatment affects the quality but not quantity of the allogeneic T cell MLR response stimulated by BCG matured DC DC were treated with 4.1 × 105 CFU/ml of BCG alone, BCG plus 1000 U/ml IFN-γ or left untreated (immature) for 24 h prior to mixed leukocyte culture. Varying numbers of DC were added to 2 × 105 enriched T cells in triplicate wells of a 96-well microculture plate and cultured for 5 days. The cells were pulsed with 3H-thymidine for an additional 24 h before harvesting and quantitation of incorporated radioactivity in a scintillation counter (A). Immediately preceding the 3H-thymidine pulse, culture supernatants were collected for evaluating IFN-γ secretion (B). Results shown in each panel are from a representative experiment out of three similar experiments.

Mentions: The observed increase in co-stimulatory molecules and TNF-α production suggested the potential for enhanced T cell stimulation by DC that were matured using BCG and IFN-γ as opposed to those matured with BCG alone. We tested this hypothesis by performing a mixed lymphocyte reaction. Immature DC, or DC matured with BCG alone or BCG plus IFN-γ for 24 h, were cultured with enriched allogeneic T cells. As shown in Figure 3A, the DC matured in the presence of BCG alone or BCG plus IFN-γ were both found to induce stronger T cell proliferative responses than immature DC, while there was no significant difference between DC matured in either manner. On the other hand, IFN-γ co-treated DC induced significantly greater IFN-γ secretion by T-cells responding to alloantigenic stimuli, especially at lower T-cell:DC ratios, than did DC matured in the absence of IFN-γ (Figure 3B). These data provided an initial indication that IFN-γ addition during BCG-induced maturation modulates a change in the quality but not the quantity of the ensuing T-cell response.


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 affects the quality but not quantity of the allogeneic T cell MLR response stimulated by BCG matured DC DC were treated with 4.1 × 105 CFU/ml of BCG alone, BCG plus 1000 U/ml IFN-γ or left untreated (immature) for 24 h prior to mixed leukocyte culture. Varying numbers of DC were added to 2 × 105 enriched T cells in triplicate wells of a 96-well microculture plate and cultured for 5 days. The cells were pulsed with 3H-thymidine for an additional 24 h before harvesting and quantitation of incorporated radioactivity in a scintillation counter (A). Immediately preceding the 3H-thymidine pulse, culture supernatants were collected for evaluating IFN-γ secretion (B). Results shown in each panel are from a representative experiment out of three similar experiments.
© Copyright Policy
Related In: Results  -  Collection

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Figure 3: IFN-γ treatment affects the quality but not quantity of the allogeneic T cell MLR response stimulated by BCG matured DC DC were treated with 4.1 × 105 CFU/ml of BCG alone, BCG plus 1000 U/ml IFN-γ or left untreated (immature) for 24 h prior to mixed leukocyte culture. Varying numbers of DC were added to 2 × 105 enriched T cells in triplicate wells of a 96-well microculture plate and cultured for 5 days. The cells were pulsed with 3H-thymidine for an additional 24 h before harvesting and quantitation of incorporated radioactivity in a scintillation counter (A). Immediately preceding the 3H-thymidine pulse, culture supernatants were collected for evaluating IFN-γ secretion (B). Results shown in each panel are from a representative experiment out of three similar experiments.
Mentions: The observed increase in co-stimulatory molecules and TNF-α production suggested the potential for enhanced T cell stimulation by DC that were matured using BCG and IFN-γ as opposed to those matured with BCG alone. We tested this hypothesis by performing a mixed lymphocyte reaction. Immature DC, or DC matured with BCG alone or BCG plus IFN-γ for 24 h, were cultured with enriched allogeneic T cells. As shown in Figure 3A, the DC matured in the presence of BCG alone or BCG plus IFN-γ were both found to induce stronger T cell proliferative responses than immature DC, while there was no significant difference between DC matured in either manner. On the other hand, IFN-γ co-treated DC induced significantly greater IFN-γ secretion by T-cells responding to alloantigenic stimuli, especially at lower T-cell:DC ratios, than did DC matured in the absence of IFN-γ (Figure 3B). These data provided an initial indication that IFN-γ addition during BCG-induced maturation modulates a change in the quality but not the quantity of the ensuing T-cell response.

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.