A novel assay system for macrophage-activating factor activity using a human U937 cell line

Mami Ishikawa; Takahiro Inoue; Toshio Inui; Daisuke Kuchiike; Kentaro Kubo; Yoshihiro Uto; Takahito Nishikata

A novel assay system for macrophage-activating factor activity using a human U937 cell line

Anticancer Res. 2014 Aug;34(8):4577-81.

Authors

Mami Ishikawa¹, Takahiro Inoue¹, Toshio Inui², Daisuke Kuchiike³, Kentaro Kubo⁴, Yoshihiro Uto⁵, Takahito Nishikata⁶

Affiliations

¹ Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, Kobe, Japan.
² Department of Life System, Institute of Technology and Science, Graduate School, University of Tokushima, Tokushima, Japan Saisei Mirai Cell Processing Center, Osaka, Japan Kobe Saisei Mirai Clinic, Kobe, Japan Inui Immunotherapy Clinic, Osaka, Japan.
³ Department of Life System, Institute of Technology and Science, Graduate School, University of Tokushima, Tokushima, Japan Saisei Mirai Cell Processing Center, Osaka, Japan.
⁴ Saisei Mirai Cell Processing Center, Osaka, Japan.
⁵ Department of Life System, Institute of Technology and Science, Graduate School, University of Tokushima, Tokushima, Japan.
⁶ Frontiers of Innovative Research in Science and Technology (FIRST), Konan University, Kobe, Japan Frontier Institute for Biomolecular Engineering Research (FIBER), Konan University, Kobe, Japan [email protected].

PMID: 25075102

Abstract

Background: Macrophages play important roles in antitumor immunity, and immunotherapy with the group-specific component protein-derived macrophage-activating factor (GcMAF) has been reported to be effective in patients with various types of cancers. However, in macrophage research, it is important to properly evaluate macrophage activity.

Materials and methods: U937 macrophages were induced by 12-O-tetradecanoyl-13-phorbolacetate (TPA). The phagocytic activity of macrophages was evaluated as the internalized beads ratio. The MAF activity was assessed at 30 min after MAF addition as the activation ratio.

Results: We established a novel assay for phagocytic activities using differentiated U937 macrophages.

Conclusion: The novel protocol was simple and rapid and was sensitive for GcMAF. This protocol should be useful not only for basic studies, such as those on molecular mechanisms underlying macrophage activation, but also for clinical studies, such as assessment of GcMAF activity prior to clinical use.

Keywords: Gc protein-derived macrophage-activating factor; GcMAF; MAF; U937 cell line; macrophage-activating factor; phagocytic activity.

MeSH terms

Dose-Response Relationship, Drug
Humans
Macrophage Activation / drug effects
Macrophage-Activating Factors / pharmacology*
Macrophages / drug effects
Macrophages / immunology
Phagocytosis / drug effects
Tetradecanoylphorbol Acetate / pharmacology
U937 Cells
Vitamin D-Binding Protein / pharmacology*

Substances

Macrophage-Activating Factors
Vitamin D-Binding Protein
vitamin D-binding protein-macrophage activating factor
Tetradecanoylphorbol Acetate