Sna 1

Ac 5 3F ax Neu5Ac was synthesized as described earlier. 20, 21 Carbo-free blocking solution and the biotinylated lectins MALII, SNA-I and PNA were obtained from Vector Laboratories, Inc. (Burlingame, CA, USA). Streptavidin-PE from BD Pharmingen (Franklin Lakes, NJ, USA), eFluor 780 viability dye from eBioscience, Inc. (San Diego, CA, USA), Lymphoprep from ELITech Group (Puteaux, France), IL-4 and GM-CSF from Cellgenix (Freiburg, Germany), methyl-3 H-thymidine (2 Ci mM) from MP Biomedicals (Costa Mesa, CA, USA). Poly(I:C) (polyinosinic-polycytidylic acid) was obtained from Enzo Life Sciences (Farmingdale, NY, USA) and LPS (lipopolysaccharide) from Sigma-Aldrich (St Louis, MO, USA). Mouse anti-human CD80-PE-Cy7 (L307.4), anti-CD86-PE-Cy7 (2331), anti-CD274-PE (MIH1) and goat anti-mouse Ig-PE were purchased from BD Bioscience (Franklin Lakes, NJ), anti-Siglec-2-PE (MEM-01) from Sigma-Aldrich, anti-Siglec-3-PE (WM53), anti-Siglec-5-PE (1A5), anti-Siglec-7-PE (6-434) and anti-Siglec-10-PE (5G6) from Biolegend, anti-Siglec-9-PE (191240) and recombinant human Siglec-His and Siglec-Fc chimera were purchased from R&D Systems (Minneapolis, MN, USA) and Alexa Fluor 647-conjugated goat anti-human IgG and mouse anti-His (HIS.H8) antibody from Thermo Scientific (Waltham, MA, USA).