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materials and methods
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Fidler (The University of Texas MD Anderson Cancer Center, Houston, TX) and described previously (van Elsas et al., >>1999<<). The prostate cancer cell line TRAMP C2 was maintained as described previously (Foster et al., 1997). B16/F10, B16/BL6, and TRAMP C2 were transduced with retrovirus to express full-length mouse ICOSL on the cell membrane and tested for
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The prostate cancer cell line TRAMP C2 was maintained as described previously (Foster et al., >>1997<<). B16/F10, B16/BL6, and TRAMP C2 were transduced with retrovirus to express full-length mouse ICOSL on the cell membrane and tested for stable expression throughout the treatment process.
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results
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In light of the significant up-regulation of ICOS on intratumoral CD8 T cells, we developed a strategy to activate the ICOS pathway by transducing tumor cells with the cognate ligand, ICOSL (Yoshinaga et al., >>1999<<), and using the irradiated ICOSL-positive tumor cells as a vaccine (IVAX) to treat tumor-bearing mice.
n2:mentions
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We have previously shown that anti–CTLA-4, when combined with GM-CSF–secreting cellular vaccine (GVAX), increases the ratio of CD8 T cells to Treg cells in the tumor (Quezada et al., >>2006<<). To evaluate the impact of IVAX and CTLA-4 blockade on the intratumoral cell composition, we counted the absolute numbers of CD8, CD4 Foxp3−, and CD4 Foxp3+ T cells in B16/F10 tumors on day 14 and normalized these numbers by the tumor
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For example, it has been shown that ectopic expression of ICOSL could elicit tumor-specific T cell response, but antitumor responses could only be generated in a prophylactic and not a therapeutic setting (Liu et al., >>2001<<; Wallin et al., 2001; Zuberek et al., 2003).
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example, it has been shown that ectopic expression of ICOSL could elicit tumor-specific T cell response, but antitumor responses could only be generated in a prophylactic and not a therapeutic setting (Liu et al., 2001; Wallin et al., >>2001<<; Zuberek et al., 2003). Similarly, engagement of the ICOS pathway with an ICOSL-Ig fusion protein alone failed to induce rejection of poorly immunogenic tumors such as B16 melanoma (Ara et al., 2003; Zuberek et al., 2003).
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that ectopic expression of ICOSL could elicit tumor-specific T cell response, but antitumor responses could only be generated in a prophylactic and not a therapeutic setting (Liu et al., 2001; Wallin et al., 2001; Zuberek et al., >>2003<<). Similarly, engagement of the ICOS pathway with an ICOSL-Ig fusion protein alone failed to induce rejection of poorly immunogenic tumors such as B16 melanoma (Ara et al., 2003; Zuberek et al., 2003).
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Similarly, engagement of the ICOS pathway with an ICOSL-Ig fusion protein alone failed to induce rejection of poorly immunogenic tumors such as B16 melanoma (Ara et al., >>2003<<; Zuberek et al., 2003).
n2:mentions
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Similarly, engagement of the ICOS pathway with an ICOSL-Ig fusion protein alone failed to induce rejection of poorly immunogenic tumors such as B16 melanoma (Ara et al., 2003; Zuberek et al., >>2003<<). This was probably caused by the fact that in the absence of CTLA-4 blockade, ICOS was expressed by only a few CD8 and/or CD4 Teff cells but expressed highly by the majority of Treg cells (Fig. 1), so that engagement of the ICOS pathway
n2:mentions
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This hypothesis was supported by the fact that pretreatment of mice with cyclophosphamide, which preferentially depleted Treg cells, could help improve the efficacy of ICOSL-Ig against poorly immunogenic tumors (Ara et al., >>2003<<). A recent study showed that human melanoma cells expressing high levels of ICOSL facilitated expansion and IL-10 production in the Treg cell population in the setting of high-dose IL-2 treatment (Martin-Orozco et al., 2010).
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A recent study showed that human melanoma cells expressing high levels of ICOSL facilitated expansion and IL-10 production in the Treg cell population in the setting of high-dose IL-2 treatment (Martin-Orozco et al., >>2010<<). In our study, however, there was no expansion of Treg cells in mice treated with IVAX alone or with anti–CTLA-4, but the balance between effector and regulatory T cells in the tumor was enhanced considerably by the combination of IVAX
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CTLA-4 blockade has been reported to selectively reduce the frequency and number of Treg cells in B16 tumors but not in the peripheral lymphoid organs (Quezada et al., >>2006<<; Simpson et al., 2013). The depletion is strongest in the combination of GVAX and anti–CTLA-4 on adoptively transferred TCR-transgenic Trp1 CD4 cells when compared with GVAX monotherapy.
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CTLA-4 blockade has been reported to selectively reduce the frequency and number of Treg cells in B16 tumors but not in the peripheral lymphoid organs (Quezada et al., 2006; Simpson et al., >>2013<<). The depletion is strongest in the combination of GVAX and anti–CTLA-4 on adoptively transferred TCR-transgenic Trp1 CD4 cells when compared with GVAX monotherapy.
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This effect is potentially driven by heavy infiltration of macrophages in the tumor (Simpson et al., >>2013<<), which is a hallmark of GVAX therapy but not observed in the IVAX model.
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The underlying mechanism of IVAX therapy is likely to be distinct from GVAX and Flt3L-secreting vaccine (FVAX; Curran and Allison, >>2009<<). An optimal cancer immunotherapy entails ample tumor antigen presentation, co-stimulation, and/or removal of co-inhibition on T cells.
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