Scrapie protein degradation by cysteine proteases in CD11c+ dendritic cells and GT1-1 neuronal cells.
Dendritic cells (DC) of the CD11c(+) myeloid phenotype have been implicated in the spread of scrapie in the host. Previously, we have shown that CD11c(+) DC can cause a rapid degradation of proteinase K-resistant prion proteins (PrP(Sc)) in vitro, indicating a possible role of these cells in the clearance of PrP(Sc). To determine the mechanisms of PrP(Sc) degradation, CD11c(+) DC that had been exposed to PrP(Sc) derived from a neuronal cell line (GT1-1) infected with scrapie (ScGT1-1) were treated with a battery of protease inhibitors. Following treatment with the cysteine protease inhibitors (2S,3S)-trans-epoxysuccinyl-L-leucylamido-3-methylbutane (E-64c), its ethyl ester (E-64d), and leupeptin, the degradation of PrP(Sc) was inhibited, while inhibitors of serine and aspartic and metalloproteases (aprotinin, pepstatin, and phosphoramidon) had no effect. An endogenous degradation of PrP(Sc) in ScGT1-1 cells was revealed by inhibiting the expression of cellular PrP (PrP(C)) by RNA interference, and this degradation could also be inhibited by the cysteine protease inhibitors. Our data show that PrP(Sc) is proteolytically cleaved preferentially by cysteine proteases in both CD11c(+) DC and ScGT1-1 cells and that the degradation of PrP(Sc) by proteases is different from that of PrP(C). Interference by protease inhibitors with DC-induced processing of PrP(Sc) has the potential to modify prion spread, clearance, and immunization in a host.
Luhr
Katarina M
KM
Department of Neuroscience, Karolinska Institutet, SE-171 77 Stockholm, Sweden. katarina.luhr@neuro.ki.se
Nordström
Elin K
EK
Löw
Peter
P
Ljunggren
Hans-Gustaf
HG
Taraboulos
Albert
A
Kristensson
Krister
K
D000818:Animals; D039521:CD11c Antigen / Q000378:metabolism*; D002460:Cell Line; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D003713:Dendritic Cells / Q000378:metabolism*; D006062:Gonadotropins / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D051379:Mice; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000378:metabolism*; D018620:PrPSc Proteins / Q000187:drug effects / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D012608:Scrapie / Q000378:metabolism
D039521:CD11c Antigen; D006062:Gonadotropins; D018620:PrPSc Proteins; D011480:Protease Inhibitors; D003546:Cysteine Endopeptidases