Fig. 1

An anti-ERMAP mAb neutralizes T cell inhibitory activity of ERMAP-Ig and enhances macrophage phagocytosis of Aβ in vitro. A T cells were purified from splenocytes of C57BL/6 mice and cultured with anti-CD3 Ab (1 μg/ml) in the presence of control Ig or human ERMAP-Ig (0.3 µg/ml) with anti-ERMAP mAb (2.5, 5, 10 µg/ml) or isotype Ab (10 µg/ml) for 3 days. [3H] thymidine (1 μCi/well) was added to the cultures 12 h before harvest. T cell proliferation was measured by [3H] thymidine incorporation. Results are expressed as counts per minute (CPM). (B, C) Splenic cells were cultured with anti-CD3 Ab in the presence of control Ig or human ERMAP-Ig (0.3 µg/ml) with anti-hERMAP mAb or control Ab (5 µg/ml). T cells were analyzed for the expression of CD69 24 h later. B Representative flow cytometric and C statistical analysis of the percentage of CD69⁺cells in CD4 and CD8 T cells. D, E Macrophages were generated from BM of C57BL/6 mice and cultured with HiLyte Fluor 647-Aβ42 in the presence of anti-hERMAP mAb or isotype Ab (5, 10 µg/ml) for 2 h. D Representative flow cytometric profiles and E statistical analysis for the percentage of HiLyte Fluor 647-Aβ42⁺ cells in F4/80⁺ macrophages. The data are expressed as mean + SD and representative of 3 independent experiments with similar results. *P < 0.05 compared with isotype Ab