A cell-free assay to estimate the neutralizing capacity of granulocyte-macrophage colony-stimulating factor autoantibodies

作者：Urano Shinya; Kaneko Chinatsu; Nei Takahito; Motoi Natsuki; Tazawa Ryushi; Watanabe Masato; Tomita Masahiro; Adachi Takahiro; Kanazawa Hiroko; Nakata Koh^*

来源：Journal of Immunological Methods, 2010, 360(1-2): 141-148.

DOI：10.1016/j.jim.2010.07.001

• GM-CSF
• GM-CSF autoantibody
• GM-CSF receptor alpha
• Receptor binding assay
• Cell-free system

The aim of the project is to develop a novel method estimating granulocyte-macrophage colony-stimulating factor (GM-CSF) neutralizing capacity with high-throughput and good reproducibility. For that purpose, we designed a cell-free receptor binding assay consisting of a solid-phase recombinant soluble GM-CSF receptor alpha (GMR alpha) and a biotinylated GM-CSF (bGM-CSF). Using this system, competitive inhibition of bGM-CSF binding to soluble GM-CSF receptor alpha (sGMR alpha) by GM-CSF autoantibody or IgG fractions from the sera of patients with pulmonary alveolar proteinosis was examined, resulting in excellent reproducibility. Binding inhibition was correlated with growth inhibition of TF-1 cells, a GM-CSF dependent cell line. These results suggest that our cell-free system can be applied to estimate the neutralizing capacity of GM-CSF autoantibodies ex vivo.

• 出版日期2010-8-31