TNF-α and Tumor Lysate Promote the Maturation of Dendritic Cells for Immunotherapy for Advanced Malignant Bone and Soft Tissue Tumors

Shinji Miwa,Munetomo Takata,Toshiharu Shirai,Hiroyuki Tsuchiya,Katsuhiro Hayashi,Akihiko Takeuchi,Hiroaki Kimura,Hideji Nishida,Norio Yamamoto,Shuichi Kaneko,Kentaro Igarashi,Yoshikazu Tanzawa,Eishiro Mizukoshi,Yasunari Nakamoto,Isaac Yang

doi:10.1371/journal.pone.0052926

Abstract

BackgroundDendritic cells (DCs) play a pivotal role in the immune system. There are many reports concerning DC-based immunotherapy. The differentiation and maturation of DCs is a critical part of DC-based immunotherapy. We investigated the differentiation and maturation of DCs in response to various stimuli.MethodsThirty-one patients with malignant bone and soft tissue tumors were enrolled in this study. All the patients had metastatic tumors and/or recurrent tumors. Peripheral blood mononuclear cells (PBMCs) were suspended in media containing interleukin-4 (IL-4) and granulocyte-macrophage colony stimulating factor (GM-CSF). These cells were then treated with or without 1) tumor lysate (TL), 2) TL + TNF-α, 3) OK-432. The generated DCs were mixed and injected in the inguinal or axillary region. Treatment courses were performed every week and repeated 6 times. A portion of the cells were analyzed by flow cytometry to determine the degree of differentiation and maturation of the DCs. Serum IFN-γ and serum IL-12 were measured in order to determine the immune response following the DC-based immunotherapy.ResultsApproximately 50% of PBMCs differentiated into DCs. Maturation of the lysate-pulsed DCs was slightly increased. Maturation of the TL/TNF-α-pulsed DCs was increased, commensurate with OK-432-pulsed DCs. Serum IFN-γ and serum IL-12 showed significant elevation at one and three months after DC-based immunotherapy.ConclusionsAlthough TL-pulsed DCs exhibit tumor specific immunity, TL-pulsed cells showed low levels of maturation. Conversely, the TL/TNF-α-pulsed DCs showed remarkable maturation. The combination of IL-4/GM-CSF/TL/TNF-α resulted in the greatest differentiation and maturation for DC-based immunotherapy for patients with bone and soft tissue tumors.

Highlights

Dendritic cells (DCs) play a pivotal role in the immune system, bridging the innate and adaptive immune responses [1,2,3]
The cells treated with IL-4, granulocyte-macrophage colony stimulating factor (GM-CSF), Tumor lysates (TL), and tumor necrotic factor alpha (TNF-a) showed more differentiation than the cells treated with IL-4, GM-CSF, and lysate alone (P,0.05). These results suggest that TL, TNF-a and OK-432 promoted the differentiation of DCs
There was no significant difference between TL group and control group. These results indicate that TL slightly induced the maturation of the DCs and that OK-432 markedly induced the maturation of DCs

Summary

Introduction

Dendritic cells (DCs) play a pivotal role in the immune system, bridging the innate and adaptive immune responses [1,2,3]. DCs are critical professional antigen-presenting cells (APCs), crucially important in the capture, processing and presenting of tumor antigens to tumor-specific T cells [4,5]. Necrotic tumor debris contains endogenous ‘‘danger signals’’, required for the activation and maturation of APCs. DCs reach the damaged tissue, take up tumor antigens in the context of inflammation and abundant cytokines, and undergo a change in phenotype characterized by the upregulation of cell surface markers, such as costimulatory, adhesion and integrin molecules. Dendritic cells (DCs) play a pivotal role in the immune system. The differentiation and maturation of DCs is a critical part of DC-based immunotherapy. We investigated the differentiation and maturation of DCs in response to various stimuli

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