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Endostatin gene therapy enhances the efficacy of IL-2 in suppressing metastatic renal cell carcinoma in mice

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Abstract

We investigated whether the administration of IL-2 combined with endostatin gene therapy was able to produce additive or even synergistic immunomodulatory activity in a mouse model of metastatic renal carcinoma. Renca cells were injected into the tail vein of BALB/c mice. After 24 h, the animals were randomly divided into four groups (5 mice/group). One group of mice was the control, the second group received treatment with 100,000 UI of Recombinant IL-2 (Proleukin, Chiron) twice a day, 1 day per week during 2 weeks (IL-2), the third group received treatment with a subcutaneous inoculation of 3.6 × 106 endostatin-producing cells, and the fourth group received both therapies (IL-2 + ES). Mice were treated for 2 weeks. In the survival studies, 10 mice/group daily, mice were monitored daily until they died. The presence of metastases led to a twofold increase in endostatin levels. Subcutaneous inoculation of NIH/3T3-LendSN cells resulted in a 2.75 and 2.78-fold increase in endostatin levels in the ES and IL-2 + ES group, respectively. At the end of the study, there was a significant decrease in lung wet weight, lung nodules area, and microvascular area (MVA) in all treated groups compared with the control group (P < 0.001). The significant difference in lung wet weight and lung nodules area between groups IL-2 and IL-2 + ES revealed a synergistic antitumor effect of the combined treatment (P < 0.05). The IL-2 + ES therapy Kaplan–Meier survival curves showed that the probability of survival was significantly higher for mice treated with the combined therapy (log-rank test, P = 0.0028). Conjugated therapy caused an increase in the infiltration of CD4, CD8 and CD49b lymphocytes. An increase in the amount of CD8 cells (P < 0.01) was observed when animals received both ES and IL-2, suggesting an additive effect of ES over IL-2 treatment. A synergistic effect of ES on the infiltration of CD4 (P < 0.001) and CD49b cells (P < 0.01) was also observed over the effect of IL-2. Here, we show that ES led to an increase in CD4 T helper cells as well as cytotoxic lymphocytes, such as NK cells and CD8 cells, within tumors of IL-2 treated mice. This means that ES plays a role in supporting the actions of T cells.

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Acknowledgments

The authors wish to thank Camila Longo Machado, PhD, for her technical help. Study supported by FAPESP (Process: 2007/54253-6).

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Authors and Affiliations

  1. Nephrology Division, Medicine Department, Federal University of São Paulo, São Paulo, Brazil

    Flávia Gomes de Góes Rocha, Karen Cristina Barbosa Chaves, Nestor Schor & Maria Helena Bellini

  2. Department of Radiology, University of São Paulo, São Paulo, Brazil

    Roger Chammas

  3. Immunology Department, Institute of Biomedical Sciences, USP, São Paulo, Brazil

    Jean Pierre Schatzmann Peron

  4. Albert Einstein Jewish Institute for Education and Research, São Paulo, Brazil

    Luiz Vicente Rizzo

  5. Biotechnology Department, IPEN-CNEN, São Paulo, SP, Brazil

    Maria Helena Bellini

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  1. Flávia Gomes de Góes Rocha
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  2. Karen Cristina Barbosa Chaves
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Correspondence to Maria Helena Bellini.

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de Góes Rocha, F.G., Chaves, K.C.B., Chammas, R. et al. Endostatin gene therapy enhances the efficacy of IL-2 in suppressing metastatic renal cell carcinoma in mice. Cancer Immunol Immunother 59, 1357–1365 (2010). https://doi.org/10.1007/s00262-010-0865-6

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