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Cooperative B7-1/2 (CD80/CD86) and B7-DC costimulation of CD4+ T cells independent of the PD-1 receptor.

Shin T, Kennedy G, Gorski K, Tsuchiya H, Koseki H, Azuma M, Yagita H, Chen L, Powell J, Pardoll D, Housseau F - J. Exp. Med. (2003)

Bottom Line: B7-1/B7-2-deficient DCs, while strongly diminished in their ability to stimulate naive CD4+ T cells, nonetheless retain partial activity.B7-DC costimulates expression of CD40L with faster kinetics than B7-1 and displays potent synergy with B7-1 and B7-2 for T cell proliferation and cytokine production, indicating that these B7 family members work in concert to stimulate T cells.Finally, costimulation with B7-DC alone or in conjunction with B7-1 is PD-1 independent, indicating that B7-DC costimulates T cells via a second receptor.

View Article: PubMed Central - PubMed

Affiliation: Sidneu Kimmel Comprehensive Cancer Centre, Johns Hopkins School of Medicine, Baltimore, MD 21231, USA.

ABSTRACT
B7-DC is a recently discovered member of the B7 family that binds to PD-1 and is selectively expressed by dendritic cells (DCs). It has been shown to either costimulate or inhibit T cell responses. To assess the role of B7-DC in DC-T cell interactions, DCs from B7-DC knockout (KO) mice were generated and compared with DCs from wild-type (WT) and B7-1/B7-2 double KO mice. B7-1/B7-2-deficient DCs, while strongly diminished in their ability to stimulate naive CD4+ T cells, nonetheless retain partial activity. DCs from B7-DC KO mice are diminished in their ability to activate CD4+ T cells, demonstrating that DC-expressed B7-DC serves a predominantly stimulatory rather than inhibitory function in the initiation of T cell responses. B7-DC costimulates expression of CD40L with faster kinetics than B7-1 and displays potent synergy with B7-1 and B7-2 for T cell proliferation and cytokine production, indicating that these B7 family members work in concert to stimulate T cells. Finally, costimulation with B7-DC alone or in conjunction with B7-1 is PD-1 independent, indicating that B7-DC costimulates T cells via a second receptor.

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HA110–120 peptide presentation to specific HA-specific CD4+ T cells by RENCA cells engineered to express B7-DC and/or B7–1 molecules. CD4+ T cells were purified from either fresh (left panels) or preactivated (right panels) splenocytes from TCR transgenic 6.5 mice (top panels) or 6.5 RAG KO mice (bottom panels). For the preactivated cells, splenocytes were cultured 4 d in vitro with 1 μg/ml of HA 110–120 peptide. CD4+ T cells were then negatively selected and rested overnight. The indicated RENCA transfectants were treated 3 d with 10 ng/ml of IFN-γ to induce expression of I-Ed molecules and used to present HA peptide (10 μg/ml) to the CD4+ T cells. T cell proliferation with either 0 (filled bars) or 10 μg/ml (open bars) HA peptide was measured after 72 h by [3H]thymidine incorporation.
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fig3: HA110–120 peptide presentation to specific HA-specific CD4+ T cells by RENCA cells engineered to express B7-DC and/or B7–1 molecules. CD4+ T cells were purified from either fresh (left panels) or preactivated (right panels) splenocytes from TCR transgenic 6.5 mice (top panels) or 6.5 RAG KO mice (bottom panels). For the preactivated cells, splenocytes were cultured 4 d in vitro with 1 μg/ml of HA 110–120 peptide. CD4+ T cells were then negatively selected and rested overnight. The indicated RENCA transfectants were treated 3 d with 10 ng/ml of IFN-γ to induce expression of I-Ed molecules and used to present HA peptide (10 μg/ml) to the CD4+ T cells. T cell proliferation with either 0 (filled bars) or 10 μg/ml (open bars) HA peptide was measured after 72 h by [3H]thymidine incorporation.

Mentions: As an alternative approach to individually reconstitute specific costimulatory signals, we transfected RENCA cells with the B7–1 gene, B7-DC gene, or both. RENCA, a BALB/c-derived tumor does not naturally express B7–1, B7–2, B7-DC, or B7-H1 even after IFN-γ treatment. IFN-γ–treated RENCA and its various transfectants express I-Ed, allowing it to present HA110–120 peptide to 6.5 transgenic CD4+ T cells. Fig. 3 shows that the optimal stimulation of either unactivated 6.5 transgenic CD4+ T cells required the presence of costimulatory molecules. In all cases, the presentation of HA110–120 by RENCA cells transfected with the combination of B7–1 and B7-DC induced the greatest T cell proliferative responses. Similarly, B7–2 + B7-DC double transfectants induced significantly greater proliferative responses among 6.5 CD4 T cells than did B7–2 or B7-DC single transfectants (unpublished data). To assess the potential of the transfectants to stimulate truly naive T cells, we used T cells 6.5 transgenic mice with RAG KO background. In contrast with RAG+/+ 6.5 transgenic splenocytes, 6.5 transgenic RAG−/− T cells are extremely dependent on costimulatory signals whereas they are much less costimulation dependent when preactivated. Whereas naive 6.5 transgenic RAG−/− CD4+ T cells were activated neither by plain RENCA nor B7–1 or B7-DC transfectants, they proliferated significantly in response to HA peptide presented by B7–1/B7-DC double transfectants.


Cooperative B7-1/2 (CD80/CD86) and B7-DC costimulation of CD4+ T cells independent of the PD-1 receptor.

Shin T, Kennedy G, Gorski K, Tsuchiya H, Koseki H, Azuma M, Yagita H, Chen L, Powell J, Pardoll D, Housseau F - J. Exp. Med. (2003)

HA110–120 peptide presentation to specific HA-specific CD4+ T cells by RENCA cells engineered to express B7-DC and/or B7–1 molecules. CD4+ T cells were purified from either fresh (left panels) or preactivated (right panels) splenocytes from TCR transgenic 6.5 mice (top panels) or 6.5 RAG KO mice (bottom panels). For the preactivated cells, splenocytes were cultured 4 d in vitro with 1 μg/ml of HA 110–120 peptide. CD4+ T cells were then negatively selected and rested overnight. The indicated RENCA transfectants were treated 3 d with 10 ng/ml of IFN-γ to induce expression of I-Ed molecules and used to present HA peptide (10 μg/ml) to the CD4+ T cells. T cell proliferation with either 0 (filled bars) or 10 μg/ml (open bars) HA peptide was measured after 72 h by [3H]thymidine incorporation.
© Copyright Policy
Related In: Results  -  Collection

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fig3: HA110–120 peptide presentation to specific HA-specific CD4+ T cells by RENCA cells engineered to express B7-DC and/or B7–1 molecules. CD4+ T cells were purified from either fresh (left panels) or preactivated (right panels) splenocytes from TCR transgenic 6.5 mice (top panels) or 6.5 RAG KO mice (bottom panels). For the preactivated cells, splenocytes were cultured 4 d in vitro with 1 μg/ml of HA 110–120 peptide. CD4+ T cells were then negatively selected and rested overnight. The indicated RENCA transfectants were treated 3 d with 10 ng/ml of IFN-γ to induce expression of I-Ed molecules and used to present HA peptide (10 μg/ml) to the CD4+ T cells. T cell proliferation with either 0 (filled bars) or 10 μg/ml (open bars) HA peptide was measured after 72 h by [3H]thymidine incorporation.
Mentions: As an alternative approach to individually reconstitute specific costimulatory signals, we transfected RENCA cells with the B7–1 gene, B7-DC gene, or both. RENCA, a BALB/c-derived tumor does not naturally express B7–1, B7–2, B7-DC, or B7-H1 even after IFN-γ treatment. IFN-γ–treated RENCA and its various transfectants express I-Ed, allowing it to present HA110–120 peptide to 6.5 transgenic CD4+ T cells. Fig. 3 shows that the optimal stimulation of either unactivated 6.5 transgenic CD4+ T cells required the presence of costimulatory molecules. In all cases, the presentation of HA110–120 by RENCA cells transfected with the combination of B7–1 and B7-DC induced the greatest T cell proliferative responses. Similarly, B7–2 + B7-DC double transfectants induced significantly greater proliferative responses among 6.5 CD4 T cells than did B7–2 or B7-DC single transfectants (unpublished data). To assess the potential of the transfectants to stimulate truly naive T cells, we used T cells 6.5 transgenic mice with RAG KO background. In contrast with RAG+/+ 6.5 transgenic splenocytes, 6.5 transgenic RAG−/− T cells are extremely dependent on costimulatory signals whereas they are much less costimulation dependent when preactivated. Whereas naive 6.5 transgenic RAG−/− CD4+ T cells were activated neither by plain RENCA nor B7–1 or B7-DC transfectants, they proliferated significantly in response to HA peptide presented by B7–1/B7-DC double transfectants.

Bottom Line: B7-1/B7-2-deficient DCs, while strongly diminished in their ability to stimulate naive CD4+ T cells, nonetheless retain partial activity.B7-DC costimulates expression of CD40L with faster kinetics than B7-1 and displays potent synergy with B7-1 and B7-2 for T cell proliferation and cytokine production, indicating that these B7 family members work in concert to stimulate T cells.Finally, costimulation with B7-DC alone or in conjunction with B7-1 is PD-1 independent, indicating that B7-DC costimulates T cells via a second receptor.

View Article: PubMed Central - PubMed

Affiliation: Sidneu Kimmel Comprehensive Cancer Centre, Johns Hopkins School of Medicine, Baltimore, MD 21231, USA.

ABSTRACT
B7-DC is a recently discovered member of the B7 family that binds to PD-1 and is selectively expressed by dendritic cells (DCs). It has been shown to either costimulate or inhibit T cell responses. To assess the role of B7-DC in DC-T cell interactions, DCs from B7-DC knockout (KO) mice were generated and compared with DCs from wild-type (WT) and B7-1/B7-2 double KO mice. B7-1/B7-2-deficient DCs, while strongly diminished in their ability to stimulate naive CD4+ T cells, nonetheless retain partial activity. DCs from B7-DC KO mice are diminished in their ability to activate CD4+ T cells, demonstrating that DC-expressed B7-DC serves a predominantly stimulatory rather than inhibitory function in the initiation of T cell responses. B7-DC costimulates expression of CD40L with faster kinetics than B7-1 and displays potent synergy with B7-1 and B7-2 for T cell proliferation and cytokine production, indicating that these B7 family members work in concert to stimulate T cells. Finally, costimulation with B7-DC alone or in conjunction with B7-1 is PD-1 independent, indicating that B7-DC costimulates T cells via a second receptor.

Show MeSH