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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 Foxp3+ Treg cells and F4/80+ macrophages in DTA-1-treated tumors.Frozen sections of CT26/NY-ESO-1 tumors obtained at 6 hrs after DTA hrs after DTA-1 i.t. injection were stained with FITC-conjugated anti-Foxp3 mAb, PE-conjugated anti-rat IgG2b mAbs and DAPI (A and B: D3), or FITC-anti-F4/80 mAb, PE-anti-rat IgG2b mAb, and DAPI (B: D1 and D2). Sections from untreated and control rat IgG-treated tumors were used as controls (A and B: N1, N2, C1, C2). Representative photos from three experiments are shown.
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pone-0104669-g005: Kinetics of Foxp3+ Treg cells and F4/80+ macrophages in DTA-1-treated tumors.Frozen sections of CT26/NY-ESO-1 tumors obtained at 6 hrs after DTA hrs after DTA-1 i.t. injection were stained with FITC-conjugated anti-Foxp3 mAb, PE-conjugated anti-rat IgG2b mAbs and DAPI (A and B: D3), or FITC-anti-F4/80 mAb, PE-anti-rat IgG2b mAb, and DAPI (B: D1 and D2). Sections from untreated and control rat IgG-treated tumors were used as controls (A and B: N1, N2, C1, C2). Representative photos from three experiments are shown.

Mentions: Fluorescent immunohistological studies using double labeling with anti-rat IgG2b mAb (for DTA-1) and F4/80- (for macrophages) or Foxp3- (for Tregs) specific mAbs were performed to determine the mechanisms of DTA-1-dependent depletion of CT26/NY-ESO-1 tumor-infiltrating Treg cells. At 6 hrs after DTA-1 treatment, Foxp3+ cells clustered at the DTA-1-stained peritumor sites, whereas Foxp3+ cells did not accumulate in the control rat IgG-treated case (Fig. 5A and S2A). Images of red fluorescence from DTA-1 or the control rat IgG merged with the green fluorescence from F4/80+ macrophages in nearby tumor stroma (Fig. 5B; C1, D1, and S2B) indicated that DTA-1 and rat IgG bound with macrophage-expressing FcRs. In addition, a large number of cells visualized in lymphocyte-like formation by staining with anti-rat IgG2b mAb were positive for Foxp3 (Fig. 5B; D2, D3; Fig. S3A and B) and were in contact with macrophages in various areas of DTA-1–treated tumors (Fig. 5B; D2; Fig. S3A). These results strongly support the hypothesis that DTA-1 participates in GITR+ Foxp3+ Treg depletion by ADCC at the treated tumor sites.


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 Foxp3+ Treg cells and F4/80+ macrophages in DTA-1-treated tumors.Frozen sections of CT26/NY-ESO-1 tumors obtained at 6 hrs after DTA hrs after DTA-1 i.t. injection were stained with FITC-conjugated anti-Foxp3 mAb, PE-conjugated anti-rat IgG2b mAbs and DAPI (A and B: D3), or FITC-anti-F4/80 mAb, PE-anti-rat IgG2b mAb, and DAPI (B: D1 and D2). Sections from untreated and control rat IgG-treated tumors were used as controls (A and B: N1, N2, C1, C2). Representative photos from three experiments are shown.
© Copyright Policy
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4126744&req=5

pone-0104669-g005: Kinetics of Foxp3+ Treg cells and F4/80+ macrophages in DTA-1-treated tumors.Frozen sections of CT26/NY-ESO-1 tumors obtained at 6 hrs after DTA hrs after DTA-1 i.t. injection were stained with FITC-conjugated anti-Foxp3 mAb, PE-conjugated anti-rat IgG2b mAbs and DAPI (A and B: D3), or FITC-anti-F4/80 mAb, PE-anti-rat IgG2b mAb, and DAPI (B: D1 and D2). Sections from untreated and control rat IgG-treated tumors were used as controls (A and B: N1, N2, C1, C2). Representative photos from three experiments are shown.
Mentions: Fluorescent immunohistological studies using double labeling with anti-rat IgG2b mAb (for DTA-1) and F4/80- (for macrophages) or Foxp3- (for Tregs) specific mAbs were performed to determine the mechanisms of DTA-1-dependent depletion of CT26/NY-ESO-1 tumor-infiltrating Treg cells. At 6 hrs after DTA-1 treatment, Foxp3+ cells clustered at the DTA-1-stained peritumor sites, whereas Foxp3+ cells did not accumulate in the control rat IgG-treated case (Fig. 5A and S2A). Images of red fluorescence from DTA-1 or the control rat IgG merged with the green fluorescence from F4/80+ macrophages in nearby tumor stroma (Fig. 5B; C1, D1, and S2B) indicated that DTA-1 and rat IgG bound with macrophage-expressing FcRs. In addition, a large number of cells visualized in lymphocyte-like formation by staining with anti-rat IgG2b mAb were positive for Foxp3 (Fig. 5B; D2, D3; Fig. S3A and B) and were in contact with macrophages in various areas of DTA-1–treated tumors (Fig. 5B; D2; Fig. S3A). These results strongly support the hypothesis that DTA-1 participates in GITR+ Foxp3+ Treg depletion by ADCC at the treated tumor sites.

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