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Silencing B7-H1 enhances the anti-tumor effect of bladder cancer antigen-loaded dendritic cell vaccine in vitro.

Wang S, Wang Y, Liu J, Shao S, Li X, Gao J, Niu H, Wang X - Onco Targets Ther (2014)

Bottom Line: After that, the infected DCs were pulsed by tumor antigens and used to stimulate cytotoxic T lymphocytes-based anti-tumor effect in vitro.The lentivirus-mediated shRNA delivery method efficiently and effectively silenced B7-H1 in DCs.This study suggests that a combination of B7-H1 knockdown and target antigen delivery could augment anti-tumor effects in vitro, which potentially provides a novel strategy in the immunotherapy of bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, The Affiliated Hospital of Qingdao University, Qingdao, People's Republic of China.

ABSTRACT

Objective: The aim of this study was to examine whether short hairpin RNA (shRNA) expressing lentiviral particles targeting B7-H1 infection could result in B7-H1 knockdown on dendritic cells (DCs) and to investigate whether B7-H1 silencing could augment the immune function of DCs and further elicit a more potent anti-tumor immune effect against bladder cancer cells in vitro.

Methods: Monocyte-derived DCs, which were generated from peripheral blood mononuclear cells, were infected by a recombinant lentivirus containing shRNA sequence aimed at B7-H1. After that, the infected DCs were pulsed by tumor antigens and used to stimulate cytotoxic T lymphocytes-based anti-tumor effect in vitro.

Results: The lentivirus-mediated shRNA delivery method efficiently and effectively silenced B7-H1 in DCs. Furthermore, the B7-H1 silencing enhanced the stimulatory capacity and the secretion of interleukin-12, but down-regulated interleukin-10 secretion. And more importantly, the anti-tumor effect of bladder cancer antigen-loaded DC vaccine in vitro was also potentially augmented.

Conclusion: This study suggests that a combination of B7-H1 knockdown and target antigen delivery could augment anti-tumor effects in vitro, which potentially provides a novel strategy in the immunotherapy of bladder cancer.

No MeSH data available.


Related in: MedlinePlus

B7-H1 silencing enhances Th1 type cytokine secretion and stimulatory capacity of DCs.Notes: (A) ELISA detection of IL-10 and IL-12 production in DCs. (B) Stimulatory capacity in mixed lymphocyte reaction. The supernatants were collected and cytokines were analyzed by ELISA. B7-H1 knockdown evidently increased IL-12 production and reduced IL-10 secretion simultaneously. T-cells’ proliferation was detected by CCK-8 kit (Dojindo, Tokyo, Japan). The result showed B7-H1-silenced DC elicited a stronger T-cell expansion in MLR. Assays were conducted in triplicate. Data were shown as mean ± standard deviation. *P<0.05 versus untreated DC.Abbreviations: IL, interleukin; NC/DC, LV-GFP-NC infected DC; shB7-H1/DC, LV-GFP-shB7-H1 infected DC; DC, dendritic cell; MLR, mixed lymphocyte reaction.
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f4-ott-7-1389: B7-H1 silencing enhances Th1 type cytokine secretion and stimulatory capacity of DCs.Notes: (A) ELISA detection of IL-10 and IL-12 production in DCs. (B) Stimulatory capacity in mixed lymphocyte reaction. The supernatants were collected and cytokines were analyzed by ELISA. B7-H1 knockdown evidently increased IL-12 production and reduced IL-10 secretion simultaneously. T-cells’ proliferation was detected by CCK-8 kit (Dojindo, Tokyo, Japan). The result showed B7-H1-silenced DC elicited a stronger T-cell expansion in MLR. Assays were conducted in triplicate. Data were shown as mean ± standard deviation. *P<0.05 versus untreated DC.Abbreviations: IL, interleukin; NC/DC, LV-GFP-NC infected DC; shB7-H1/DC, LV-GFP-shB7-H1 infected DC; DC, dendritic cell; MLR, mixed lymphocyte reaction.

Mentions: To investigate whether B7-H1 silencing could improve the immune function of DC vaccine, the capacity to stimulate T-cell proliferation was measured by mixed lymphocyte reaction and cytokine productions were analyzed by ELISA. We found that DC vaccine combined with B7-H1 silencing demonstrated a stronger capability to stimulate T-cell proliferation. Meanwhile, IL-12 secretion was increased and IL-10 secretion was decreased after B7-H1 silencing. Thus, our results indicate that B7-H1 silencing can enhance the stimulatory capacity and Th1 type cytokine secretion of DC vaccine (Figure 4).


Silencing B7-H1 enhances the anti-tumor effect of bladder cancer antigen-loaded dendritic cell vaccine in vitro.

Wang S, Wang Y, Liu J, Shao S, Li X, Gao J, Niu H, Wang X - Onco Targets Ther (2014)

B7-H1 silencing enhances Th1 type cytokine secretion and stimulatory capacity of DCs.Notes: (A) ELISA detection of IL-10 and IL-12 production in DCs. (B) Stimulatory capacity in mixed lymphocyte reaction. The supernatants were collected and cytokines were analyzed by ELISA. B7-H1 knockdown evidently increased IL-12 production and reduced IL-10 secretion simultaneously. T-cells’ proliferation was detected by CCK-8 kit (Dojindo, Tokyo, Japan). The result showed B7-H1-silenced DC elicited a stronger T-cell expansion in MLR. Assays were conducted in triplicate. Data were shown as mean ± standard deviation. *P<0.05 versus untreated DC.Abbreviations: IL, interleukin; NC/DC, LV-GFP-NC infected DC; shB7-H1/DC, LV-GFP-shB7-H1 infected DC; DC, dendritic cell; MLR, mixed lymphocyte reaction.
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4128850&req=5

f4-ott-7-1389: B7-H1 silencing enhances Th1 type cytokine secretion and stimulatory capacity of DCs.Notes: (A) ELISA detection of IL-10 and IL-12 production in DCs. (B) Stimulatory capacity in mixed lymphocyte reaction. The supernatants were collected and cytokines were analyzed by ELISA. B7-H1 knockdown evidently increased IL-12 production and reduced IL-10 secretion simultaneously. T-cells’ proliferation was detected by CCK-8 kit (Dojindo, Tokyo, Japan). The result showed B7-H1-silenced DC elicited a stronger T-cell expansion in MLR. Assays were conducted in triplicate. Data were shown as mean ± standard deviation. *P<0.05 versus untreated DC.Abbreviations: IL, interleukin; NC/DC, LV-GFP-NC infected DC; shB7-H1/DC, LV-GFP-shB7-H1 infected DC; DC, dendritic cell; MLR, mixed lymphocyte reaction.
Mentions: To investigate whether B7-H1 silencing could improve the immune function of DC vaccine, the capacity to stimulate T-cell proliferation was measured by mixed lymphocyte reaction and cytokine productions were analyzed by ELISA. We found that DC vaccine combined with B7-H1 silencing demonstrated a stronger capability to stimulate T-cell proliferation. Meanwhile, IL-12 secretion was increased and IL-10 secretion was decreased after B7-H1 silencing. Thus, our results indicate that B7-H1 silencing can enhance the stimulatory capacity and Th1 type cytokine secretion of DC vaccine (Figure 4).

Bottom Line: After that, the infected DCs were pulsed by tumor antigens and used to stimulate cytotoxic T lymphocytes-based anti-tumor effect in vitro.The lentivirus-mediated shRNA delivery method efficiently and effectively silenced B7-H1 in DCs.This study suggests that a combination of B7-H1 knockdown and target antigen delivery could augment anti-tumor effects in vitro, which potentially provides a novel strategy in the immunotherapy of bladder cancer.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, The Affiliated Hospital of Qingdao University, Qingdao, People's Republic of China.

ABSTRACT

Objective: The aim of this study was to examine whether short hairpin RNA (shRNA) expressing lentiviral particles targeting B7-H1 infection could result in B7-H1 knockdown on dendritic cells (DCs) and to investigate whether B7-H1 silencing could augment the immune function of DCs and further elicit a more potent anti-tumor immune effect against bladder cancer cells in vitro.

Methods: Monocyte-derived DCs, which were generated from peripheral blood mononuclear cells, were infected by a recombinant lentivirus containing shRNA sequence aimed at B7-H1. After that, the infected DCs were pulsed by tumor antigens and used to stimulate cytotoxic T lymphocytes-based anti-tumor effect in vitro.

Results: The lentivirus-mediated shRNA delivery method efficiently and effectively silenced B7-H1 in DCs. Furthermore, the B7-H1 silencing enhanced the stimulatory capacity and the secretion of interleukin-12, but down-regulated interleukin-10 secretion. And more importantly, the anti-tumor effect of bladder cancer antigen-loaded DC vaccine in vitro was also potentially augmented.

Conclusion: This study suggests that a combination of B7-H1 knockdown and target antigen delivery could augment anti-tumor effects in vitro, which potentially provides a novel strategy in the immunotherapy of bladder cancer.

No MeSH data available.


Related in: MedlinePlus