2aG4 directly induces monocytic MDSCs to differentiate into dendritic cells and macrophages by binding to phosphatidylserine on their cell surface. 2aG4.

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2aG4 directly induces monocytic MDSCs to differentiate into dendritic cells and macrophages by binding to phosphatidylserine on their cell surface. 2aG4 directly induces monocytic MDSCs to differentiate into dendritic cells and macrophages by binding to phosphatidylserine on their cell surface. A, FACS analyses showing that monocytic MDSCs (CD11b+, Ly6Glo, and Ly6Chi) from the spleens of tumor-bearing mice differentiate into dendritic cells and macrophages when cultured for 4 days with 2aG4. MDSCs (CD11b+ and Gr-1+), macrophages (F4/80+ and Gr-1−), and dendritic cells (CDIIb+, CD11chi, and Gr-1−) were quantified by FACS. Histograms, mean percentage ± SD. Statistically significant for 2aG4 versus C44 (P < 0.0001; Student two-tailed t test; three determinations). B, treatment of monocytic MDSCs with 2aG4 induces NO production (Griess assay) in an Fc-dependent manner. Histograms, mean ± SD of triplicate determinations. C, ELISAs showing that 2aG4 treatment of monocytic MDSCs induces the production of inflammatory cytokines. D, qRT-PCR showing that 2aG4 treatment of monocytic MDSCs causes them to acquire markers of M1 macrophages and mature dendritic cells. The mRNA levels in 2aG4-treated cells are expressed relative to those in C44-treated cells. E, FACS analysis showing that monocytic MDSCs have exposed phosphatidylserine. F, TEM of a monocytic MDSC showing that phosphatidylserine on the cell surface is due to the presence of phosphatidylserine-positive microvesicles (100–500 nm diameter; Alix-). Arrows, gold-labeled 2aG4. Gold-labeled C44 did not bind (not shown). Scale bars, black, 1,000 nm; white, 200 nm. Yi Yin et al. Cancer Immunol Res 2013;1:256-268 ©2013 by American Association for Cancer Research