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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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Generation of tumor- and HF10-specific CD8+ T cells by intratumoral treatment of DTA-1 and HF10 combined with DTA-1.(A) CT26- and NY-ESO-1-specific CD8+ T cell responses in CT26/NY-ESO-1-regressed mice by i.t. treatment of DTA-1 at indicated doses were assessed by intracellular staining of IFN-γ in splenocytes cultured with 10 µg/mL of the indicated peptides for 5 hrs. Splenocytes from two mice per group were pooled and assessed. hrs. Splenocytes from two mice per group were pooled and assessed. (B) Splenocytes from untreated and both i.t. HF10- and DTA-1-treated CT26/NY-ESO-1-bearing mice were obtained at 5 days after final treatment, and cultured with AH-1 peptide (10 µg/mL) or HF10-infected CMS5a tumor cells for 5 hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN-γ. The 9 m peptide and uninfected CMS5a cells were used as controls. m peptide and uninfected CMS5a cells were used as controls.
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pone-0104669-g003: Generation of tumor- and HF10-specific CD8+ T cells by intratumoral treatment of DTA-1 and HF10 combined with DTA-1.(A) CT26- and NY-ESO-1-specific CD8+ T cell responses in CT26/NY-ESO-1-regressed mice by i.t. treatment of DTA-1 at indicated doses were assessed by intracellular staining of IFN-γ in splenocytes cultured with 10 µg/mL of the indicated peptides for 5 hrs. Splenocytes from two mice per group were pooled and assessed. hrs. Splenocytes from two mice per group were pooled and assessed. (B) Splenocytes from untreated and both i.t. HF10- and DTA-1-treated CT26/NY-ESO-1-bearing mice were obtained at 5 days after final treatment, and cultured with AH-1 peptide (10 µg/mL) or HF10-infected CMS5a tumor cells for 5 hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN-γ. The 9 m peptide and uninfected CMS5a cells were used as controls. m peptide and uninfected CMS5a cells were used as controls.

Mentions: Next, we investigated whether the tumor- or HF10-specific CD8+ T cell response was enhanced in CT26/NY-ESO-1-bearing mice by i.t. treatment with DTA-1 alone or HF10 combined with DTA-1. To detect low proportions of CD8+ T cells with tumor specificity, spleen cells from tumor-regressed mice selected after i.t. treatment with DTA-1 at the indicated doses were stimulated with a CT26-specific AH-1 peptide or an NY-ESO-1 81–88 peptide to expand each population of peptide-specific CD8+ T cells. As shown in Figure 3A, the response of CT26-specific IFN-γ-producing CD8+ T cells was enhanced by DTA-1 in a dose-dependent manner. In addition, CD8+ T cells with NY-ESO-1 specificity were observed when DTA-1 was administered at a high dose (10 µg). In this experiment, we used tumor-regressed mice because we could not enhance the negligible CT26-specific IFN-γ-producing CD8+ T cell responses seen in tumor-bearers in a DTA-1 dose-dependent manner. Although CT26/NY-ESO-1 growth after i.t. treatment with DTA-1 was suppressed compared with the control group, tumor size was not different in each group of DTA-1 (0.5, 2.0, or 10.0 µg/mouse). Furthermore, HF10-specific CD8+ T cells were found in addition to the AH-1-specific population (Fig. 3B left) when splenocytes from both HF10- and DTA-1-treated CT26/NY-ESO-1-bearing mice with HF10-infected CMS5a cells were cultured (Fig. 3B right). These results indicated that i.t. treatment of HF10 and DTA-1 had the capacity to enhance tumor- and HF10-specific CD8+ T cell populations.


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)

Generation of tumor- and HF10-specific CD8+ T cells by intratumoral treatment of DTA-1 and HF10 combined with DTA-1.(A) CT26- and NY-ESO-1-specific CD8+ T cell responses in CT26/NY-ESO-1-regressed mice by i.t. treatment of DTA-1 at indicated doses were assessed by intracellular staining of IFN-γ in splenocytes cultured with 10 µg/mL of the indicated peptides for 5 hrs. Splenocytes from two mice per group were pooled and assessed. hrs. Splenocytes from two mice per group were pooled and assessed. (B) Splenocytes from untreated and both i.t. HF10- and DTA-1-treated CT26/NY-ESO-1-bearing mice were obtained at 5 days after final treatment, and cultured with AH-1 peptide (10 µg/mL) or HF10-infected CMS5a tumor cells for 5 hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN-γ. The 9 m peptide and uninfected CMS5a cells were used as controls. m peptide and uninfected CMS5a cells were used as controls.
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4126744&req=5

pone-0104669-g003: Generation of tumor- and HF10-specific CD8+ T cells by intratumoral treatment of DTA-1 and HF10 combined with DTA-1.(A) CT26- and NY-ESO-1-specific CD8+ T cell responses in CT26/NY-ESO-1-regressed mice by i.t. treatment of DTA-1 at indicated doses were assessed by intracellular staining of IFN-γ in splenocytes cultured with 10 µg/mL of the indicated peptides for 5 hrs. Splenocytes from two mice per group were pooled and assessed. hrs. Splenocytes from two mice per group were pooled and assessed. (B) Splenocytes from untreated and both i.t. HF10- and DTA-1-treated CT26/NY-ESO-1-bearing mice were obtained at 5 days after final treatment, and cultured with AH-1 peptide (10 µg/mL) or HF10-infected CMS5a tumor cells for 5 hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN hrs. Splenocytes from ten mice per group were pooled and assessed. The obtained cells were immunohistologically stained for intracellular IFN-γ. The 9 m peptide and uninfected CMS5a cells were used as controls. m peptide and uninfected CMS5a cells were used as controls.
Mentions: Next, we investigated whether the tumor- or HF10-specific CD8+ T cell response was enhanced in CT26/NY-ESO-1-bearing mice by i.t. treatment with DTA-1 alone or HF10 combined with DTA-1. To detect low proportions of CD8+ T cells with tumor specificity, spleen cells from tumor-regressed mice selected after i.t. treatment with DTA-1 at the indicated doses were stimulated with a CT26-specific AH-1 peptide or an NY-ESO-1 81–88 peptide to expand each population of peptide-specific CD8+ T cells. As shown in Figure 3A, the response of CT26-specific IFN-γ-producing CD8+ T cells was enhanced by DTA-1 in a dose-dependent manner. In addition, CD8+ T cells with NY-ESO-1 specificity were observed when DTA-1 was administered at a high dose (10 µg). In this experiment, we used tumor-regressed mice because we could not enhance the negligible CT26-specific IFN-γ-producing CD8+ T cell responses seen in tumor-bearers in a DTA-1 dose-dependent manner. Although CT26/NY-ESO-1 growth after i.t. treatment with DTA-1 was suppressed compared with the control group, tumor size was not different in each group of DTA-1 (0.5, 2.0, or 10.0 µg/mouse). Furthermore, HF10-specific CD8+ T cells were found in addition to the AH-1-specific population (Fig. 3B left) when splenocytes from both HF10- and DTA-1-treated CT26/NY-ESO-1-bearing mice with HF10-infected CMS5a cells were cultured (Fig. 3B right). These results indicated that i.t. treatment of HF10 and DTA-1 had the capacity to enhance tumor- and HF10-specific CD8+ T cell populations.

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