Cd169

Paraffin-embedded and formalin-fixed HCC samples were cut into 5-μm sections, which were then processed for immunohistochemistry as previously described. Briefly, the sections were incubated with Abs against the following human antigens: CD8 (rabbit monoclonal, clone EP334, ZSBio, China), CD11b (rabbit monoclonal, clone EPR1344, Abcam, UK), CD68 (mouse monoclonal; clone PG-M1; Dako Cytomation, USA), CD15 (mouse monoclonal; clone LeuM1; ZSBio), CD206 (mouse monoclonal, clone 685645, R&D Systems, USA), CD204 (mouse monoclonal, clone SRA-C6, Transgenic, Japan), CD163 (mouse monoclonal, clone 10D6, ZSBio) and CD169 (sheep polyclonal, clone NS0, R&D Systems). Then, the sections were stained in an EnVision System (Dako Cytomation, USA). Positive cells were detected by microscopy and quantified using the Vectra-Inform image analysis system (Perkin-Elmer Applied Biosystems, Foster City, CA, USA).

Paraffin-embedded and formalin-fixed HCC samples were cut into 5-μm sections, which were then processed for immunohistochemistry as previously described. Briefly, the sections were incubated with Abs against the following human antigens: CD8 (rabbit monoclonal, clone EP334, ZSBio, China), CD11b (rabbit monoclonal, clone EPR1344, Abcam, UK), CD68 (mouse monoclonal; clone PG-M1; Dako Cytomation, USA), CD15 (mouse monoclonal; clone LeuM1; ZSBio), CD206 (mouse monoclonal, clone 685645, R&D Systems, USA), CD204 (mouse monoclonal, clone SRA-C6, Transgenic, Japan), CD163 (mouse monoclonal, clone 10D6, ZSBio) and CD169 (sheep polyclonal, clone NS0, R&D Systems). Then, the sections were stained in an EnVision System (Dako Cytomation, USA). Positive cells were detected by microscopy and quantified using the Vectra-Inform image analysis system (Perkin-Elmer Applied Biosystems, Foster City, CA, USA).

The thymus and spleen immunofluorescence was performed using a previously described method^{43 (link)}. Sections were immunostained with the antibodies against CD3, CD4, CD8 (all from eBioscience), CD169 (R&D System), UEA-1 (Vector Laboratories), and ER-TR7 (AbD Serotec) and agitated overnight at 4 °C. Stained thick sections were microscopically analyzed using a Leica SP5 confocal microscope (Leica Microsystem, Inc.) and images were processed with Leica LAS AF software (Leica Microsystem, Inc.) and Imaris v.7.7.1 64x (Bitplane).