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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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Disappearance of DTA-1-conjugated tumor-infiltrating CD4+ Foxp3+ cells after combined i.t. treatment with HF10 and DTA-1.(A) CT26/NY-ESO-1 tumor sections form untreated group (control) or group from mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with FITC-anti-Foxp3 mAb and DAPI. (B) The frequency of tumor-infiltrating CD4+ Foxp3+ Treg cells from mice injected i.t. with HF10, DTA-1, or HF10 combined with DTA-1 at 12 days after CT26/NY-ESO-1 inoculation was assessed by flow cytometry. Data from 4 individual experiments were analyzed statistically. Kruskal-Wallis ANOVA test was used to compare data from the 4 groups. The decrease in the frequency of tumoral CD4+ Foxp3+ Treg cells in the HF10 and DTA-1 combined treatment group was significantly different from untreated control, but not from the HF10- or DTA-1-treated group (N.S: Not significant). (C) At 6 hrs after DTA hrs after DTA-1 injection into day 9 CT26/NY-ESO-1 tumors, tumor-infiltrating cells collected under collagenase-free conditions were analyzed by flow cytometry after staining with FITC-labeled anti-rat IgG2b mAb to detect DTA-1-bound cells.
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pone-0104669-g004: Disappearance of DTA-1-conjugated tumor-infiltrating CD4+ Foxp3+ cells after combined i.t. treatment with HF10 and DTA-1.(A) CT26/NY-ESO-1 tumor sections form untreated group (control) or group from mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with FITC-anti-Foxp3 mAb and DAPI. (B) The frequency of tumor-infiltrating CD4+ Foxp3+ Treg cells from mice injected i.t. with HF10, DTA-1, or HF10 combined with DTA-1 at 12 days after CT26/NY-ESO-1 inoculation was assessed by flow cytometry. Data from 4 individual experiments were analyzed statistically. Kruskal-Wallis ANOVA test was used to compare data from the 4 groups. The decrease in the frequency of tumoral CD4+ Foxp3+ Treg cells in the HF10 and DTA-1 combined treatment group was significantly different from untreated control, but not from the HF10- or DTA-1-treated group (N.S: Not significant). (C) At 6 hrs after DTA hrs after DTA-1 injection into day 9 CT26/NY-ESO-1 tumors, tumor-infiltrating cells collected under collagenase-free conditions were analyzed by flow cytometry after staining with FITC-labeled anti-rat IgG2b mAb to detect DTA-1-bound cells.

Mentions: We hypothesized that the increase in tumor-specific CD8+ T cell responses after DTA-1 treatment combined with HF10 therapy was involved in the attenuation and/or depletion of immune suppressors including Treg cells. To address this issue, CT26/NY-ESO-1 tumors obtained after DTA-1 treatment with or without HF10 were evaluated by immunohistological staining of tissue sections and flow cytometric analysis of infiltrating lymphocytes using a Foxp3-specific mAb. Foxp3+ cells accumulated abundantly in both untreated and HF10-treated tumors, whereas a vigorous decrease in the number of Treg cells was shown in tumors following treatment with DTA-1 alone or DTA-1 combined with HF10 (Fig. 4A). This result was confirmed by flow cytometric analysis of tumor-infiltrating Treg cells. The frequency of tumoral CD4+ Foxp3+ Treg cells from the HF10- and DTA-1-treated group was decreased significantly compared with that from the untreated (control) group but not from the HF10- or DTA-1- treated mice (Fig. 4B). The decrease in the frequency of Foxp3+ cells in the HF10-treated group (Fig. 4B) is possibly correlated with the decrease in tumor size due to HF10 treatment. This may be attributed to the lack of modulation of the absolute number of Foxp3+ cells in the HF10-treated group unlike in the control group (Fig. 4A). DTA-1 is a rat IgG2b class mAb. By visualizing DTA-1 with the FITC-conjugated anti-rat IgG2b mAb, it was demonstrated that the tumor-infiltrating CD4+ Foxp3+ Treg population bound predominantly with DTA-1 at 6 hrs after i.t. injection (Fig. 4C), in parallel with the disappearance of tumoral Treg cells after treatment with DTA-1. Taken together, these results strongly indicate that DTA-1 was essential to the decrease in the number of CD4+ Foxp3+ cells.


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)

Disappearance of DTA-1-conjugated tumor-infiltrating CD4+ Foxp3+ cells after combined i.t. treatment with HF10 and DTA-1.(A) CT26/NY-ESO-1 tumor sections form untreated group (control) or group from mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with FITC-anti-Foxp3 mAb and DAPI. (B) The frequency of tumor-infiltrating CD4+ Foxp3+ Treg cells from mice injected i.t. with HF10, DTA-1, or HF10 combined with DTA-1 at 12 days after CT26/NY-ESO-1 inoculation was assessed by flow cytometry. Data from 4 individual experiments were analyzed statistically. Kruskal-Wallis ANOVA test was used to compare data from the 4 groups. The decrease in the frequency of tumoral CD4+ Foxp3+ Treg cells in the HF10 and DTA-1 combined treatment group was significantly different from untreated control, but not from the HF10- or DTA-1-treated group (N.S: Not significant). (C) At 6 hrs after DTA hrs after DTA-1 injection into day 9 CT26/NY-ESO-1 tumors, tumor-infiltrating cells collected under collagenase-free conditions were analyzed by flow cytometry after staining with FITC-labeled anti-rat IgG2b mAb to detect DTA-1-bound cells.
© Copyright Policy
Related In: Results  -  Collection

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

pone-0104669-g004: Disappearance of DTA-1-conjugated tumor-infiltrating CD4+ Foxp3+ cells after combined i.t. treatment with HF10 and DTA-1.(A) CT26/NY-ESO-1 tumor sections form untreated group (control) or group from mice injected i.t. with DTA-1, HF10, or HF10 combined with DTA-1 were stained with FITC-anti-Foxp3 mAb and DAPI. (B) The frequency of tumor-infiltrating CD4+ Foxp3+ Treg cells from mice injected i.t. with HF10, DTA-1, or HF10 combined with DTA-1 at 12 days after CT26/NY-ESO-1 inoculation was assessed by flow cytometry. Data from 4 individual experiments were analyzed statistically. Kruskal-Wallis ANOVA test was used to compare data from the 4 groups. The decrease in the frequency of tumoral CD4+ Foxp3+ Treg cells in the HF10 and DTA-1 combined treatment group was significantly different from untreated control, but not from the HF10- or DTA-1-treated group (N.S: Not significant). (C) At 6 hrs after DTA hrs after DTA-1 injection into day 9 CT26/NY-ESO-1 tumors, tumor-infiltrating cells collected under collagenase-free conditions were analyzed by flow cytometry after staining with FITC-labeled anti-rat IgG2b mAb to detect DTA-1-bound cells.
Mentions: We hypothesized that the increase in tumor-specific CD8+ T cell responses after DTA-1 treatment combined with HF10 therapy was involved in the attenuation and/or depletion of immune suppressors including Treg cells. To address this issue, CT26/NY-ESO-1 tumors obtained after DTA-1 treatment with or without HF10 were evaluated by immunohistological staining of tissue sections and flow cytometric analysis of infiltrating lymphocytes using a Foxp3-specific mAb. Foxp3+ cells accumulated abundantly in both untreated and HF10-treated tumors, whereas a vigorous decrease in the number of Treg cells was shown in tumors following treatment with DTA-1 alone or DTA-1 combined with HF10 (Fig. 4A). This result was confirmed by flow cytometric analysis of tumor-infiltrating Treg cells. The frequency of tumoral CD4+ Foxp3+ Treg cells from the HF10- and DTA-1-treated group was decreased significantly compared with that from the untreated (control) group but not from the HF10- or DTA-1- treated mice (Fig. 4B). The decrease in the frequency of Foxp3+ cells in the HF10-treated group (Fig. 4B) is possibly correlated with the decrease in tumor size due to HF10 treatment. This may be attributed to the lack of modulation of the absolute number of Foxp3+ cells in the HF10-treated group unlike in the control group (Fig. 4A). DTA-1 is a rat IgG2b class mAb. By visualizing DTA-1 with the FITC-conjugated anti-rat IgG2b mAb, it was demonstrated that the tumor-infiltrating CD4+ Foxp3+ Treg population bound predominantly with DTA-1 at 6 hrs after i.t. injection (Fig. 4C), in parallel with the disappearance of tumoral Treg cells after treatment with DTA-1. Taken together, these results strongly indicate that DTA-1 was essential to the decrease in the number of CD4+ Foxp3+ cells.

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