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Systemic CD8+ T cell-mediated tumoricidal effects by intratumoral treatment of oncolytic herpes simplex virus with the agonistic monoclonal antibody for murine glucocorticoid-induced tumor necrosis factor receptor.

Ishihara M, Seo N, Mitsui J, Muraoka D, Tanaka M, Mineno J, Ikeda H, Shiku H - PLoS ONE (2014)

Bottom Line: In this study, we examined the tumoricidal effects of oncolytic HF10, a naturally occurring mutant of herpes simplex virus type-1, combined with an agonistic DTA-1 monoclonal antibody specific for the glucocorticoid-induced tumor necrosis factor receptor.The kinetics and immunological mechanisms of DTA-1 in HF10 infection were examined using flow cytometry and immunohistochemistry.Studies using Fc-digested DTA-1 and Fcγ receptor knockout mice demonstrated the direct participation of DTA-1 in regulatory T cell depletion by antibody-dependent cellular cytotoxicity primarily via macrophages.

View Article: PubMed Central - PubMed

Affiliation: Department of Immuno-Gene Therapy, Mie University Graduate School of Medicine, Mie, Japan.

ABSTRACT
Oncolytic virotherapy combined with immunomodulators is a novel noninvasive strategy for cancer treatment. In this study, we examined the tumoricidal effects of oncolytic HF10, a naturally occurring mutant of herpes simplex virus type-1, combined with an agonistic DTA-1 monoclonal antibody specific for the glucocorticoid-induced tumor necrosis factor receptor. Two murine tumor models were used to evaluate the therapeutic efficacies of HF10 virotherapy combined with DTA-1. The kinetics and immunological mechanisms of DTA-1 in HF10 infection were examined using flow cytometry and immunohistochemistry. Intratumoral administration of HF10 in combination with DTA-1 at a low dose resulted in a more vigorous attenuation of growth of the untreated contralateral as well as the treated tumors than treatment with either HF10 or DTA-1 alone. An accumulation of CD8(+) T cells, including tumor- and herpes simplex virus type-1-specific populations, and a decrease in the number of CD4(+) Foxp3(+) T regulatory cells were seen in both HF10- and DTA-1-treated tumors. Studies using Fc-digested DTA-1 and Fcγ receptor knockout mice demonstrated the direct participation of DTA-1 in regulatory T cell depletion by antibody-dependent cellular cytotoxicity primarily via macrophages. These results indicated the potential therapeutic efficacy of a glucocorticoid-induced tumor necrosis factor receptor-specific monoclonal antibody in oncolytic virotherapy at local tumor sites.

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Kinetics of CD8+ T cells after the combination therapy with HF10 and DTA-1.CT26/NY-ESO-1 tumor sections from untreated mice (control) or mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with hematoxylin and eosin (A), and phycoerythrin (PE)-conjugated anti-CD8α mAb and DAPI (B). (C) Frozen sections of CT26/NY-ESO-1 tumors from mice i.t. injected with HF10 combined with DTA-1 were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI. (D) CT26/NY-ESO-1 growth (mm2) in both i.t. HF10- and DTA-1-treated control or CD8+ cell-depleted mice was measured. Seven mice per group were used. (E) Bilateral CT26/NY-ESO-1-bearing mice were treated with a combination of HF10 and DTA-1 in the tumors on the right flanks. Subsequent tumor growth (mm2) of the treated right and contralateral left sites was measured. Tumor growth in untreated mice was measured and used as a control. Fourteen mice per group were used. By the Kruskal-Wallis ANOVA test, CT26/NY-ESO-1 growth inhibition by the combined HF10 and DTA-1 treatment in contralateral as well as treated sites was significantly different from the untreated control group. (F) CT26/NY-ESO-1 tumors from one side of bilateral tumor-bearing mice were treated i.t. with HF10 combined with DTA-1. Frozen sections of contralateral CT26/NY-ESO-1 tumors were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI.
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pone-0104669-g002: Kinetics of CD8+ T cells after the combination therapy with HF10 and DTA-1.CT26/NY-ESO-1 tumor sections from untreated mice (control) or mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with hematoxylin and eosin (A), and phycoerythrin (PE)-conjugated anti-CD8α mAb and DAPI (B). (C) Frozen sections of CT26/NY-ESO-1 tumors from mice i.t. injected with HF10 combined with DTA-1 were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI. (D) CT26/NY-ESO-1 growth (mm2) in both i.t. HF10- and DTA-1-treated control or CD8+ cell-depleted mice was measured. Seven mice per group were used. (E) Bilateral CT26/NY-ESO-1-bearing mice were treated with a combination of HF10 and DTA-1 in the tumors on the right flanks. Subsequent tumor growth (mm2) of the treated right and contralateral left sites was measured. Tumor growth in untreated mice was measured and used as a control. Fourteen mice per group were used. By the Kruskal-Wallis ANOVA test, CT26/NY-ESO-1 growth inhibition by the combined HF10 and DTA-1 treatment in contralateral as well as treated sites was significantly different from the untreated control group. (F) CT26/NY-ESO-1 tumors from one side of bilateral tumor-bearing mice were treated i.t. with HF10 combined with DTA-1. Frozen sections of contralateral CT26/NY-ESO-1 tumors were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI.

Mentions: Intratumoral injection of HF10 resulted in the collapse of tumor structure with a decrease in the nuclear density of tumor cells, as shown at 7 days after the last HF10 treatment in both the group treated with HF10 and that treated with HF10 and DTA-1 (Fig. 2A). Tumor-infiltrating CD8+ T cells were shown to be the most frequent population after the administration of HF10 combined with DTA-1 at 3 days after the final treatment (Fig. 2B). Importantly, these cells appeared to accumulate near HF10-infected tumor areas (Fig. 2C and S1A), suggesting that HF10 infection is able to attract CD8+ T cells by leaking virus-associated proteins and tumor antigenic proteins from infected tumor cells and changing the tumor microenvironment after oncolysis. Inhibition of CT26/NY-ESO-1 growth by the combination therapy was completely negated by depleting CD8+ cells by intravenous treatment with a murine CD8α-specific mAb (Fig. 2D), indicating that the tumor-infiltrating CD8+ T cells shown in Figure 2B and 2C include tumoricidal effector populations. In the study using bilateral tumor-bearing mice, tumor growth inhibition by HF10 combined with DTA-1 occurred not only in the treated tumors but also in the contralateral non-treated tumors (Fig. 2E and S1B). In addition, the sections of contralateral tumor showed infiltrating CD8+ T cells without HF10 infection (Fig. 2F and S1C). These results indicate that CD8+ T cells activated in a local tumor site under the influence of HF10 and DTA-1 participate in systemic surveillance and could attack distant tumors without tissue destruction due to HF10 infection.


Systemic CD8+ T cell-mediated tumoricidal effects by intratumoral treatment of oncolytic herpes simplex virus with the agonistic monoclonal antibody for murine glucocorticoid-induced tumor necrosis factor receptor.

Ishihara M, Seo N, Mitsui J, Muraoka D, Tanaka M, Mineno J, Ikeda H, Shiku H - PLoS ONE (2014)

Kinetics of CD8+ T cells after the combination therapy with HF10 and DTA-1.CT26/NY-ESO-1 tumor sections from untreated mice (control) or mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with hematoxylin and eosin (A), and phycoerythrin (PE)-conjugated anti-CD8α mAb and DAPI (B). (C) Frozen sections of CT26/NY-ESO-1 tumors from mice i.t. injected with HF10 combined with DTA-1 were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI. (D) CT26/NY-ESO-1 growth (mm2) in both i.t. HF10- and DTA-1-treated control or CD8+ cell-depleted mice was measured. Seven mice per group were used. (E) Bilateral CT26/NY-ESO-1-bearing mice were treated with a combination of HF10 and DTA-1 in the tumors on the right flanks. Subsequent tumor growth (mm2) of the treated right and contralateral left sites was measured. Tumor growth in untreated mice was measured and used as a control. Fourteen mice per group were used. By the Kruskal-Wallis ANOVA test, CT26/NY-ESO-1 growth inhibition by the combined HF10 and DTA-1 treatment in contralateral as well as treated sites was significantly different from the untreated control group. (F) CT26/NY-ESO-1 tumors from one side of bilateral tumor-bearing mice were treated i.t. with HF10 combined with DTA-1. Frozen sections of contralateral CT26/NY-ESO-1 tumors were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI.
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Related In: Results  -  Collection

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pone-0104669-g002: Kinetics of CD8+ T cells after the combination therapy with HF10 and DTA-1.CT26/NY-ESO-1 tumor sections from untreated mice (control) or mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with hematoxylin and eosin (A), and phycoerythrin (PE)-conjugated anti-CD8α mAb and DAPI (B). (C) Frozen sections of CT26/NY-ESO-1 tumors from mice i.t. injected with HF10 combined with DTA-1 were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI. (D) CT26/NY-ESO-1 growth (mm2) in both i.t. HF10- and DTA-1-treated control or CD8+ cell-depleted mice was measured. Seven mice per group were used. (E) Bilateral CT26/NY-ESO-1-bearing mice were treated with a combination of HF10 and DTA-1 in the tumors on the right flanks. Subsequent tumor growth (mm2) of the treated right and contralateral left sites was measured. Tumor growth in untreated mice was measured and used as a control. Fourteen mice per group were used. By the Kruskal-Wallis ANOVA test, CT26/NY-ESO-1 growth inhibition by the combined HF10 and DTA-1 treatment in contralateral as well as treated sites was significantly different from the untreated control group. (F) CT26/NY-ESO-1 tumors from one side of bilateral tumor-bearing mice were treated i.t. with HF10 combined with DTA-1. Frozen sections of contralateral CT26/NY-ESO-1 tumors were stained with a PE-anti-CD8α monoclonal antibody, a fluorescein isothiocyanate (FITC)-anti-HSV-1 polyclonal antibody, and DAPI.
Mentions: Intratumoral injection of HF10 resulted in the collapse of tumor structure with a decrease in the nuclear density of tumor cells, as shown at 7 days after the last HF10 treatment in both the group treated with HF10 and that treated with HF10 and DTA-1 (Fig. 2A). Tumor-infiltrating CD8+ T cells were shown to be the most frequent population after the administration of HF10 combined with DTA-1 at 3 days after the final treatment (Fig. 2B). Importantly, these cells appeared to accumulate near HF10-infected tumor areas (Fig. 2C and S1A), suggesting that HF10 infection is able to attract CD8+ T cells by leaking virus-associated proteins and tumor antigenic proteins from infected tumor cells and changing the tumor microenvironment after oncolysis. Inhibition of CT26/NY-ESO-1 growth by the combination therapy was completely negated by depleting CD8+ cells by intravenous treatment with a murine CD8α-specific mAb (Fig. 2D), indicating that the tumor-infiltrating CD8+ T cells shown in Figure 2B and 2C include tumoricidal effector populations. In the study using bilateral tumor-bearing mice, tumor growth inhibition by HF10 combined with DTA-1 occurred not only in the treated tumors but also in the contralateral non-treated tumors (Fig. 2E and S1B). In addition, the sections of contralateral tumor showed infiltrating CD8+ T cells without HF10 infection (Fig. 2F and S1C). These results indicate that CD8+ T cells activated in a local tumor site under the influence of HF10 and DTA-1 participate in systemic surveillance and could attack distant tumors without tissue destruction due to HF10 infection.

Bottom Line: In this study, we examined the tumoricidal effects of oncolytic HF10, a naturally occurring mutant of herpes simplex virus type-1, combined with an agonistic DTA-1 monoclonal antibody specific for the glucocorticoid-induced tumor necrosis factor receptor.The kinetics and immunological mechanisms of DTA-1 in HF10 infection were examined using flow cytometry and immunohistochemistry.Studies using Fc-digested DTA-1 and Fcγ receptor knockout mice demonstrated the direct participation of DTA-1 in regulatory T cell depletion by antibody-dependent cellular cytotoxicity primarily via macrophages.

View Article: PubMed Central - PubMed

Affiliation: Department of Immuno-Gene Therapy, Mie University Graduate School of Medicine, Mie, Japan.

ABSTRACT
Oncolytic virotherapy combined with immunomodulators is a novel noninvasive strategy for cancer treatment. In this study, we examined the tumoricidal effects of oncolytic HF10, a naturally occurring mutant of herpes simplex virus type-1, combined with an agonistic DTA-1 monoclonal antibody specific for the glucocorticoid-induced tumor necrosis factor receptor. Two murine tumor models were used to evaluate the therapeutic efficacies of HF10 virotherapy combined with DTA-1. The kinetics and immunological mechanisms of DTA-1 in HF10 infection were examined using flow cytometry and immunohistochemistry. Intratumoral administration of HF10 in combination with DTA-1 at a low dose resulted in a more vigorous attenuation of growth of the untreated contralateral as well as the treated tumors than treatment with either HF10 or DTA-1 alone. An accumulation of CD8(+) T cells, including tumor- and herpes simplex virus type-1-specific populations, and a decrease in the number of CD4(+) Foxp3(+) T regulatory cells were seen in both HF10- and DTA-1-treated tumors. Studies using Fc-digested DTA-1 and Fcγ receptor knockout mice demonstrated the direct participation of DTA-1 in regulatory T cell depletion by antibody-dependent cellular cytotoxicity primarily via macrophages. These results indicated the potential therapeutic efficacy of a glucocorticoid-induced tumor necrosis factor receptor-specific monoclonal antibody in oncolytic virotherapy at local tumor sites.

Show MeSH
Related in: MedlinePlus