Validation of Bone Marrow Derived Dendritic Cells as an Appropriate Model to Study Tumor-Mediated Suppression of DC Maturation through STAT3 Hyperactivation

Authors

  • Aws Alshamsan University of Alberta
  • Samar Hamdy University of Alberta
  • Saswati Das Saswati Das University of Alberta
  • Afsaneh Lavasanifar
  • John Samuel University of Alberta
  • Ayman O.S. El-Kadi University of Alberta

DOI:

https://doi.org/10.18433/J37598

Abstract

PURPOSE: Tumors can escape immune eradication by harnessing dendritic cell (DC) maturation. However, DC types used as in vitro models to study tumor-mediated immunosuppression possess fundamental variability that could influence research outcomes. Therefore, we assessed the behavior of two distinct murine DC models upon exposure to tumor-conditioned medium of B16.F10 melanoma (B16-CM). METHODS: Using primary bone-marrow derived dendritic cells (BMDCs) or immortalized DC2.4 cell line, we evaluated the level of signal transducer and activator of transcription 3 (STAT3) phosphorylation by Western blot as a molecular parameter. We also examined the surface expression of co-stimulatory molecules on DCs by flow cytometry as a phenotypic parameter. RESULTS: Our results revealed critical discrepancies between the two models in response to tumor-conditioned medium. While conditioned medium was able to induce STAT3 phosphorylation in BMDCs, it did not significantly induce STAT3 phosphorylation in DC2.4 cell line. Moreover, only in BMDCs, the expression of CD86 and CD40 was remarkably downregulated by B16-CM and was not totally recovered after LPS stimulation. In contrast, DC2.4 cells did not show any signs of harnessed maturation upon exposure to B16-CM. CONCLUSIONS: In order to study the effect of tumor-mediated immunosuppression on DC maturation in vitro via tumor-induction of STAT3 activation, primary BMDCs are more reliable as a model than DC2.4.

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Author Biographies

Aws Alshamsan, University of Alberta

PhD Student

Samar Hamdy, University of Alberta

Postdoctoral fellow

Saswati Das Saswati Das, University of Alberta

Postdoctoral fellow

Afsaneh Lavasanifar

Associate Professor

John Samuel, University of Alberta

deseased

Ayman O.S. El-Kadi, University of Alberta

Associate Professor

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Published

2010-02-02

How to Cite

Alshamsan, A., Hamdy, S., Saswati Das, S. D., Lavasanifar, A., Samuel, J., & El-Kadi, A. O. (2010). Validation of Bone Marrow Derived Dendritic Cells as an Appropriate Model to Study Tumor-Mediated Suppression of DC Maturation through STAT3 Hyperactivation. Journal of Pharmacy & Pharmaceutical Sciences, 13(1), 21–26. https://doi.org/10.18433/J37598

Issue

Section

Pharmaceutical Sciences; Review Articles