TY - JOUR
T1 - Scrapie Protein Degradation by Cysteine Proteases in CD11c+ Dendritic Cells and GT1-1 Neuronal Cells
AU - Luhr, Katarina M.
AU - Nordström, Elin K.
AU - Löw, Peter
AU - Ljunggren, Hans Gustaf
AU - Taraboulos, Albert
AU - Kristensson, Krister
PY - 2004/5
Y1 - 2004/5
N2 - 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 (PrPSc) in vitro, indicating a possible role of these cells in the clearance of PrPSc. To determine the mechanisms of PrPSc degradation, CD11c+ DC that had been exposed to PrPSc 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 PrPSc was inhibited, while inhibitors of serine and aspartic and metalloproteases (aprotinin, pepstatin, and phosphoramidon) had no effect. An endogenous degradation of PrPSc in ScGT1-1 cells was revealed by inhibiting the expression of cellular PrP (PrPC) by RNA interference, and this degradation could also be inhibited by the cysteine protease inhibitors. Our data show that PrPSc is proteolytically cleaved preferentially by cysteine proteases in both CD11c+ DC and ScGT1-1 cells and that the degradation of PrPSc by proteases is different from that of PrP C. Interference by protease inhibitors with DC-induced processing of PrPSc has the potential to modify prion spread, clearance, and immunization in a host.
AB - 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 (PrPSc) in vitro, indicating a possible role of these cells in the clearance of PrPSc. To determine the mechanisms of PrPSc degradation, CD11c+ DC that had been exposed to PrPSc 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 PrPSc was inhibited, while inhibitors of serine and aspartic and metalloproteases (aprotinin, pepstatin, and phosphoramidon) had no effect. An endogenous degradation of PrPSc in ScGT1-1 cells was revealed by inhibiting the expression of cellular PrP (PrPC) by RNA interference, and this degradation could also be inhibited by the cysteine protease inhibitors. Our data show that PrPSc is proteolytically cleaved preferentially by cysteine proteases in both CD11c+ DC and ScGT1-1 cells and that the degradation of PrPSc by proteases is different from that of PrP C. Interference by protease inhibitors with DC-induced processing of PrPSc has the potential to modify prion spread, clearance, and immunization in a host.
UR - http://www.scopus.com/inward/record.url?scp=3242772435&partnerID=8YFLogxK
U2 - 10.1128/JVI.78.9.4776-4782.2004
DO - 10.1128/JVI.78.9.4776-4782.2004
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C2 - 15078959
AN - SCOPUS:3242772435
SN - 0022-538X
VL - 78
SP - 4776
EP - 4782
JO - Journal of Virology
JF - Journal of Virology
IS - 9
ER -