Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The . gov means it’s official. Federal government websites often end in . gov or . mil VSports app下载. Before sharing sensitive information, make sure you’re on a federal government site. .

Https

The site is secure. The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely. V体育官网.

. 2001 Jul;108(1):51-62.
doi: 10.1172/JCI10128.

IFN-gamma and Fas/FasL are required for the antitumor and antiangiogenic effects of IL-12/pulse IL-2 therapy

J M Wigginton  1 , E Gruys, L Geiselhart, J Subleski, K L Komschlies, J W Park, T A Wiltrout, K Nagashima, T C Back, R H Wiltrout
Affiliations

IFN-gamma and Fas/FasL are required for the antitumor and antiangiogenic effects of IL-12/pulse IL-2 therapy

J M Wigginton et al. J Clin Invest. 2001 Jul.

VSports在线直播 - Abstract

Systemic administration of IL-12 and intermittent doses of IL-2 induce complete regression of metastatic murine renal carcinoma VSports手机版. Here, we show that overt tumor regression induced by IL-12/pulse IL-2 is preceded by recruitment of CD8(+) T cells, vascular injury, disrupted tumor neovascularization, and apoptosis of both endothelial and tumor cells. The IL-12/IL-2 combination synergistically enhances cell surface FasL expression on CD8(+) T lymphocytes in vitro and induces Fas and FasL expression within tumors via an IFN-gamma-dependent mechanism in vivo. This therapy also inhibits tumor neovascularization and induces tumor regression by mechanisms that depend critically on endogenous IFN-gamma production and an intact Fas/FasL pathway. The ability of IL-12/pulse IL-2 to induce rapid destruction of tumor-associated endothelial cells and regression of established metastatic tumors is ablated in mice with a dysregulated Fas/FasL pathway. The common, critical role for endogenous IFN-gamma and the Fas/FasL pathway in early antiangiogenic effects and in antitumor responses suggests that early, cytokine-driven innate immune mechanisms and CD8(+) T cell-mediated responses are interdependent. Definition of critical early molecular events engaged by IL-12/IL-2 may provide new perspective into optimal therapeutic engagement of a productive host-antitumor immune response. .

PubMed Disclaimer

Figures

Figure 1
Figure 1
Infiltration of CD8+ T lymphocytes in established Renca tumors after treatment with IL-12/pulse IL-2. Cohorts of mice received subcutaneous Renca implants and were subsequently treated with IL-12 (0.5 μg daily on days 11–15 and 18–21), pulse IL-2 (300,000 IU twice daily on days 11 and 18), or vehicle alone. After two cycles of therapy, tumors were resected and stained for expression of CD8 or CD4. Anti-CD8 sections from control mice (c) and mice treated with IL-12/pulse IL-2 (d) and anti-CD4–stained sections from control (a) and IL-12/pulse IL-2–treated (b) mice are shown. ×200.
Figure 2
Figure 2
Detection of intracellular IFN-γ in splenic T lymphocyte subsets after in vivo treatment with IL-12/pulse IL-2. BALB/c mice were treated with IL-12 (0.5 μg daily on days 0, 1, and 2) and pulse IL-2 (300,000 IU twice daily on day 0) or vehicle alone. On day 3, single cell suspensions of splenic leukocytes were prepared, and intracellular IFN-γ expression by CD4+ and CD8+ T lymphocytes was determined by flow cytometry as described in Methods. Histograms represent the percentages of CD4+ and CD8+ T cells, respectively, which express IFN-γ after treatment with IL-12/pulse IL-2 or vehicle alone. Bar graphs represent the absolute number of cells expressing IFN-γ per spleen (×105).
Figure 3
Figure 3
Role of IFN-γ in the antitumor activity of IL-12/pulse IL-2 against metastatic Renca. WT versus IFN-γ–/– mice received intrarenal Renca tumor implants. Mice underwent unilateral nephrectomy to remove the primary tumor-bearing kidney on day 12 after tumor implantation and were subsequently treated with IL-12 (0.5 μg daily on days 13–17, 20–24, 27–31, and 34–38), pulse IL-2 (300,000 IU twice daily on days 13, 20, 27, 34, and 41), or vehicle alone. Mice surviving at last follow-up were tumor-free.
Figure 4
Figure 4
Role of IFN-γ in the antivascular activity of IL-12/pulse IL-2 against intrarenal Renca. WT versus IFN-γ–/– mice received intrarenal Renca tumor implants. Mice were treated with IL-12 (0.5 μg daily on days 11–15 and 18–20), pulse IL-2 (300,000 IU twice daily on days 11 and 18), or vehicle alone. Tumors were resected on day 21 and stained for CD31 expression. Representative sections from WT mice treated with IL-12/pulse IL-2 (b) or vehicle alone (a) and IFN-γ–/– mice treated with IL-12/pulse IL-2 (d) or vehicle alone (c) are shown. ×100.
Figure 5
Figure 5
Vascularity of advanced intrarenal Renca tumors and impact of IL-12/pulse IL-2 administration on tumor neovascularization. (a–c) The extensive vascularity of untreated advanced (3.5 weeks old) intrarenal Renca tumors was visualized as described in Methods. Using these methods, the vascularity of established intrarenal Renca tumors was compared directly after treatment of mice with IL-12 (0.5 μg daily on days 8–12 and 15–18) plus pulse IL-2 (300,000 IU twice daily on days 8 and 15) (f) or vehicle alone (d and e). Quantitative assessment of the impact of IL-12/pulse IL-2 administration on vascularization of intrarenal Renca tumor implants (g). Cohorts of mice received subcapsular intrarenal Renca tumor implants and were treated with IL-12 (0.5 μg on days 12–16 and 19–22), pulse IL-2 (300,000 IU twice daily on days 12 and 19), or vehicle alone. At baseline (day 12) and after one (day 16) or two (day 22) cycles of therapy, cohorts of mice were euthanized, and latex suspension containing 111In-ox was infused as described in Methods. The normal and tumor-bearing kidneys were resected, and the amount of radioactivity in each organ was quantitated. Values for the tumor-bearing kidney were normalized to the non–tumor-bearing kidney in each animal. Open circles represent the respective values for individual mice euthanized at the indicated time point.
Figure 6
Figure 6
Demonstration of direct correlation between increased accumulation of 111In-ox latex infusate and increased vascularity of Gelfoam sponges (control or bFGF-treated) were affixed onto the surface of the left kidney as described previously. Ten days later, mice were euthanized, infused with 111In-ox–containing liquid latex as described for Figure 5, and sponges were excised for visual inspection and quantitation of radioisotope incorporation. The results show that as expected, control sponges are poorly vascularized (a), bFGF sponges are well vascularized (b), and the degree of vascularity correlates directly with an increase in isotope accumulation (c).
Figure 7
Figure 7
Impact of IL-12/pulse IL-2 administration on local gene expression within established Renca tumors in WT versus IFN-γ–/– mice. Subcutaneous Renca tumors were resected from mice treated with IL-12 (0.5 μg daily on days 11–15 and 18–20), pulse IL-2 (300,000 IU twice daily on days 11 and 18), or vehicle alone, and gene expression was characterized by RT-PCR. Each lane represents analysis of material from an individual tumor site.
Figure 8
Figure 8
Impact of IL-12 with or without IL-2 on cell surface FasL expression by lymph node–derived lymphocytes. Activated lymph node–derived whole lymphocyte populations were cultured in vitro in the presence of KB8301 metalloproteinase inhibitor with IL-12, IL-2, or medium alone, and cell surface FasL expression was subsequently detected using immunofluorescent labeling with flow cytometric analysis as described in Methods. The respective histograms represent the fluorescent intensity distribution for cells cultured under the indicated conditions. The red and blue tracings represent fluorescence-intensity distribution histograms for cells cultured under the indicated conditions and stained subsequently with isotype control or hamster anti-mouse FasL Ab’s, respectively. The percentage of cells that were positive for FasL expression in the respective groups are indicated in each histogram.
Figure 9
Figure 9
Impact of administration of IL-12/pulse IL-2 on the ultrastructural histology of endothelial cells within established Renca tumors. Intrarenal Renca tumors were resected from mice treated with IL-12 (0.5 μg daily on days 11–14), pulse IL-2 (300,000 IU twice daily on day 11), or vehicle alone, and tissue specimens were processed for subsequent examination using electron microscopy. (a) Endothelial cells line a blood vessel within a control Renca tumor. ×6,000. (b–d) Endothelial cells from Renca tumors treated with IL-12/pulse IL-2. (b and c) ×6000. (d) ×30,000.
Figure 10
Figure 10
Role of the Fas/FasL pathway in IL-12/pulse IL-2–induced endothelial apoptosis within established renal tumors. Intrarenal tumors were resected from WT or lpr mice treated with IL-12 (0.5 μg daily on days 11–14), pulse IL-2 (300,000 IU twice daily on day 11), or vehicle alone, as indicated, and tissue specimens were processed for subsequent examination using electron microscopy. (a–c) Endothelial cells from control Renca tumors. (d–f) Endothelial cells from Renca tumors in IL-12/pulse IL-2–treated WT and (g–i) lpr mice. (a–d, f–i) ×3,000. (e) ×9,000.
Figure 11
Figure 11
Role of FasL in mediating the antitumor activity of IL-12/pulse IL-2 against metastatic Renca. Cohorts of WT versus gld mice bearing intrarenal Renca tumor implants underwent unilateral nephrectomy to remove the primary tumor-bearing kidney on day 12 after tumor implantation and were subsequently treated with IL-12 (0.5 μg daily on days 13–16, 20–24, 27–31, and 34–37), pulse IL-2 (300,000 IU twice daily on days 13, 20, 27, and 34), or vehicle alone. Mice surviving at last follow-up were tumor-free.
See this image and copyright information in PMC

VSports - References

    1. Trinchieri G. Interleukin-12: a cytokine at the interface of inflammation and immunity. Adv Immunol. 1998;70:83–243. - PubMed
    1. Brunda MJ, et al. Antitumor and antimetastatic activity of interleukin 12 against murine tumors. J Exp Med. 1993;178:1223–1230. - PMC (V体育官网) - PubMed
    1. Nastala CL, et al. Recombinant IL-12 administration induces tumor regression in association with IFN-gamma production. J Immunol. 1994;153:1697–1706. - PubMed
    1. Gately MK, Wilson DE, Wong HL. Synergy between recombinant interleukin 2 (rIL 2) and IL 2-depleted lymphokine-containing supernatants in facilitating allogeneic human cytolytic T lymphocyte responses in vitro. J Immunol. 1986;136:1274–1282. - PubMed
    1. Chan SH, et al. Induction of interferon gamma production by natural killer cell stimulatory factor: characterization of the responder cells and synergy with other inducers. J Exp Med. 1991;173:869–879. - PMC - PubMed

"VSports在线直播" Publication types

MeSH terms

"VSports" Grants and funding