Effect of reduced endocytosis induced by hypotonic shock and potassium depletion on the infection of Hep 2 cells by picornaviruses.
Potassium depletion after a brief exposure of the cells to hypotonic medium was used to inhibit endocytosis from coated pits in Hep 2 cells. After such treatment the endocytic uptake of transferrin was arrested, and electron microscopy revealed that virtually no coated pits were present at the cell surface, while smooth (uncoated) pits were abundant. Under the same conditions the cells were strongly protected against poliovirus, while the cytopathogenic effect of human rhinovirus type 2, HRV 2, was increased. The cytopathogenic effect of encephalomyocarditis (EMC) virus was only slightly affected. Potassium depletion without hypotonic shock reduced the endocytic uptake of transferrin 2-3-fold and the number of coated pits at the cell surface about 3-fold. Furthermore, the cells were not protected against poliovirus after such treatment. The data indicate that the productive uptake of poliovirus occurs by receptor-mediated endocytosis from coated pits, while the productive uptake of the other two picornaviruses may occur by another endocytic pathway. In order to efficiently arrest endocytosis from coated pits in these cells, hypotonic shock seems to be a critical component of the potassium depletion protocol.
D002460:Cell Line; D004686:Encephalomyocarditis virus / Q000378:metabolism; D004705:Endocytosis; D006801:Humans; D007038:Hypotonic Solutions; D007457:Iodine Radioisotopes / Q000378:metabolism; D007822:Laryngeal Neoplasms / Q000378:metabolism / Q000382:microbiology; D008854:Microscopy, Electron; D010849:Picornaviridae / Q000378:metabolism*; D010850:Picornaviridae Infections / Q000473:pathology; D017955:Poliovirus / Q000378:metabolism; D011188:Potassium / Q000502:physiology; D011191:Potassium Deficiency / Q000378:metabolism*; D012367:RNA, Viral / Q000378:metabolism; D012229:Rhinovirus / Q000378:metabolism; D012276:Ricin / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism
D007038:Hypotonic Solutions; D007457:Iodine Radioisotopes; D012367:RNA, Viral; D014168:Transferrin; D012276:Ricin; D011188:Potassium