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Mol Cells ; 34(3): 263-70, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22949172

ABSTRACT

BCG-activated macrophages (BAM) could kill the tumor cells through cell-cell contact. In this process membrane proteins play an important role. However, up to date, few membrane proteins were revealed. In this study, we selected a surface molecule named Trim59, which was specifically expressed on BAM membrane (compared with the negative control). We cloned and prokaryoticly expressed the extracellular domain of Trim59, purified the recombinant protein and generated polyclonal antibodies. Immunohistochemistry showed that Trim59 abundantly expressed in spleen, stomach and ovary; intermediately expressed in brain, lung, kidney, muscle and intestine; but not in thymus, liver, heart, uterus. Using the antibodies to block Trim59 on BAM significantly reduced BAM cytotoxicity against MCA207 cells. This demonstrated that Trim59 serves as an indispensable molecule in maintaining BAM activity. Overexpression of Trim59 in Raw264.7 cell line failed to lyse target MCA207 cells, which potentiated Trim59 per se could not enhance macrophage cytotoxicity; on another hand, overexpression of Trim59 enhance the pinocytosis and Phagocytosis activity of Raw-264.7, which imply Trim59 might mediate the cell-molecule interaction. Our results indicate Trim59 might be an essential accessory molecule in mediating BAM tumoricidal functions; and Trim59 is a phagocytosis-correlated molecule.


Subject(s)
Carrier Proteins/genetics , Carrier Proteins/immunology , Cytotoxicity, Immunologic/physiology , Macrophage Activation/immunology , Macrophages, Peritoneal/immunology , Mycobacterium bovis/immunology , Amino Acid Sequence , Animals , Base Sequence , Carrier Proteins/metabolism , Cells, Cultured , Cloning, Molecular , Cytotoxicity, Immunologic/immunology , Female , Gene Expression Profiling , Intracellular Signaling Peptides and Proteins , Macrophages, Peritoneal/metabolism , Mice , Mice, Inbred C57BL , Molecular Sequence Data , Phagocytosis/immunology , Tripartite Motif Proteins
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