Functional selection of phagocytosis-promoting genes: cell sorting-based selection.
Phagocytosis is a critical host defense mechanism that clears invading pathogens, apoptotic cells, and cell debris; it is an essential process for normal development, tissue remodeling, immune response, and inflammation. Here, a functional selection strategy was used to isolate novel phagocytosis-promoting genes. After the retroviral transfer of mouse brain cDNA library into NIH3T3 mouse fibroblast cells, cell sorting was used to select the cells that phagocytosed fluorescent zymosan particles. The cDNAs were retrieved from the selected cells and identified by DNA sequencing as eIF5A, Meg3, Tubb5, Sparcl-1, Uchl-1, Bsg (CD147), Ube2v1, and Pamr1. The phagocytosis-promoting activity for some of these cDNAs was confirmed by transient transfection in the independent phagocytosis assays. Thus, the unbiased selection procedure successfully identified multiple phagocytosis-promoting genes. The selection method can be applied to other cell-based assays where cells with a desired phenotype can be physically separated. Moreover, the new gene targets uncovered in this study could be relevant to biomolecule screening in search of phagocytosis-regulating agents. In a small-scale screen, a series of imidazopyridine compounds was tested to identify the small molecules that modulate eIF5A-mediated phagocytic activity. Several compounds that influenced the phagocytic activity can be further used as chemical-genetic tools to delineate the mechanisms of eIF5A action and be potential drug candidates that are capable of therapeutically modulating phagocytic activity.
Jeon
Hyejin
H
Department of Pharmacology, Brain Science and Engineering Institute, CMRI, Kyungpook National University School of Medicine, Daegu, Korea.
Go
Younghoon
Y
Seo
Minchul
M
Lee
Won-Ha
WH
Suk
Kyoungho
K
D000818:Animals; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D003001:Cloning, Molecular / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D015723:Gene Library; D005796:Genes / Q000187:drug effects / Q000502:physiology; D057166:High-Throughput Screening Assays / Q000379:methods; D051379:Mice; D008954:Models, Biological; D041681:NIH 3T3 Cells; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D010587:Phagocytosis / Q000187:drug effects / Q000235:genetics*; D010641:Phenotype; D014162:Transfection
D019384:Nucleic Acid Synthesis Inhibitors