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Effective Photoimmunotherapy of Murine Colon Carcinoma Induced by the Combination of Photodynamic Therapy and Dendritic Cells

Ahmad Jalili¹, Marcin Makowski¹, Tomasz witaj¹, Dominika Nowis¹, Grzegorz M. Wilczyski³, Ewa Wilczek³, Magdalena Chory-Massalska⁴, Anna Radzikowska⁴, Wodzimierz Maliski⁴, ukasz Biay², Jacek Sieko⁵, Aleksander Siero⁶, Mariusz Adamek⁶, Grzegorz Basak¹, Pawe Mróz¹, Ireneusz W. Krasnodbski⁷, Marek Jakóbisiak¹ and Jakub Gob¹

Departments of ¹ Immunology, ² Histology and Embryology, and ³ Pathology, Center of Biostructure Research, The Medical University of Warsaw, Warsaw; ⁴ Department of Pathophysiology and Immunology, Institute of Rheumatology, Warsaw, ⁵ II Department and Clinic of Obstetrics and Gynecology, The Medical University of Warsaw, Warsaw; ⁶ Center for Laser Diagnostics and Therapy, Chair and Clinic of Internal Diseases and Physical Medicine, Silesian Medical University, Bytom; and ⁷ Department of General, Gastrointestinal Surgery and Nutrition, The Medical University of Warsaw, Warsaw, Poland

Purpose: The unique mechanism of tumor destruction by photodynamic therapy (PDT), resulting from apoptotic and necrotic killing of tumor cells accompanied by local inflammatory reaction and induction of heat shock proteins (HSPs), prompted us to investigate the antitumor effectiveness of the combination of PDT with administration of immature dendritic cells (DCs).

Experimental Design: Confocal microscopy and Western blotting were used to investigate the influence of PDT on the induction of apoptosis and expression of HSP expression in C-26 cells. Confocal microscopy and flow cytometry studies were used to examine phagocytosis of PDT-treated C-26 cells by DCs. Secretion of interleukin (IL)-12 was measured with ELISA. Cytotoxic activity of lymph node cells was evaluated in a standard ⁵¹Cr-release assay. The antitumor effectiveness of PDT in combination with administration of DCs was investigated in in vivo model.

Results: PDT treatment resulted in the induction of apoptotic and necrotic cell death and expression of HSP27, HSP60, HSP72/73, HSP90, HO-1, and GRP78 in C-26 cells. Immature DCs cocultured with PDT-treated C-26 cells efficiently engulfed killed tumor cells, acquired functional features of maturation, and produced substantial amounts of IL-12. Inoculation of immature DCs into the PDT-treated tumors resulted in effective homing to regional and peripheral lymph nodes and stimulation of cytotoxic activity of T and natural killer cells. The combination treatment with PDT and administration of DCs produced effective antitumor response.

Conclusions: The feasibility and antitumor effectiveness demonstrated in these studies suggest that treatment protocols involving the administration of immature DCs in combination with PDT may have clinical potential.

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