Photodynamic therapy plus low-dose cyclophosphamide generates antitumor immunity in a mouse model

Photodynamic therapy plus low-dose cyclophosphamide generates antitumor immunity in a mouse model

Ana P. Castano*,,, Pawel Mroz*,, Mei X. Wu*,,, and Michael R. Hamblin*,,,¶

*Wellman Center for Photomedicine, Massachusetts General Hospital, Boston, MA 02114; Department of Dermatology, Harvard Medical School, Boston, MA 02115; and Division of Health Sciences and Technology, Harvard–Massachusetts Institute of Technology, Cambridge, MA 02139

Edited by Mark Naylor, University of Oklahoma Health Sciences Center, Tulsa, OK, and accepted by the Editorial Board February 21, 2008 (received for review September 28, 2007)

Abstract

Photodynamic therapy (PDT) is a modality for the treatment of cancer involving excitation of nontoxic photosensitizers with harmless visible light-producing cytotoxic reactive oxygen species.

PDT causes apoptosis and necrosis of tumor cells, destruction of the tumor blood supply, and activation of the immune system.

The objective of this study was to compare in an animal model of metastatic cancer PDT alone and PDT combined with low-dose cyclophosphamide (CY) a treatment that has been proposed to deplete regulatory T cells (T-regs) and increase the immune response to some tumors.

We used J774 tumors (a highly metastatic reticulum cell sarcoma line) and PDT with benzoporphyrin derivative monoacid ring A, verteporfin for injection (BPD; 1-mg/kg injected i.v. followed after 15 min by 150 J/cm2 of 690-nm light). CY (50 or 150 mg/kg i.p.) was injected 48 h before light delivery.

PDT alone led to tumor regressions and a survival advantage but no permanent cures were obtained.

BPD–PDT in combination with low-dose CY (but not high-dose CY) led to 70% permanent cures.

Low-dose CY alone gave no permanent cures but did provide a survival advantage and was shown to reduce CD4+FoxP3+ T-regs in lymph nodes, whereas high-dose CY reduced other lymphocyte classes as well.

Cured animals were rechallenged with J774 cells, and the tumors were rejected in 71% of mice.

Cured mice had tumor-specific T cells in spleens as determined by a 51Cr release assay.

We conclude that low-dose CY depletes T-regs and potentiates BPD–PDT, leading to tumor cures and memory immunity.