Leucht Christoph, Simoneau Steve, Rey Clémence, Vana Karen, Rieger Roman, Lasmézas Corinne Ida, Weiss Stefan
Laboratorium für Molekulare Biologie-Genzentrum-Institut für Biochemie der LMU München, Feodor-Lynen Str. 25, D- 81377 Munich, Germany.
EMBO Rep. 2003 Mar;4(3):290-5. doi: 10.1038/sj.embor.embor768.
The accumulation of PrP(Sc) in scrapie-infected neuronal cells has been prevented by three approaches: (i) transfection of ScMNB cells with an antisense laminin receptor precursor (LRP) RNA-expression plasmid, (ii) transfection of ScN2a cells and ScGT1 cells with small interfering RNAs (siRNAs) specific for the LRP mRNA, and (iii) incubation of ScN2a cells with an anti-LRP/LR antibody. LRP antisense RNA and LRP siRNAs reduced LRP/LR expression and inhibited the accumulation of PrP(Sc) in these cells. The treatments also reduced PrP(c) levels. The anti-LRP/LR antibody, W3, abolished PrP(Sc) accumulation and reduced PrP(c) levels after seven days of incubation. Cells remained free of PrP(Sc) after being cultured for 14 additional days without the antibody, whereas the PrP(c) level was restored. Our results demonstrate the necessity of the laminin receptor (LRP/LR) for PrP(Sc) propagation in cultured cells and suggest that LRP/LR-specific antibodies could be used as powerful therapeutic tools in the treatment of transmissible spongiform encephalopathies.
通过三种方法可防止瘙痒病感染的神经元细胞中PrP(Sc)的积累：(i) 用反义层粘连蛋白受体前体（LRP）RNA表达质粒转染ScMNB细胞；(ii) 用针对LRP mRNA的小干扰RNA（siRNA）转染ScN2a细胞和ScGT1细胞；(iii) 用抗LRP/LR抗体孵育ScN2a细胞。LRP反义RNA和LRP siRNA降低了LRP/LR的表达，并抑制了这些细胞中PrP(Sc)的积累。这些处理也降低了PrP(c)的水平。抗LRP/LR抗体W3在孵育7天后消除了PrP(Sc)的积累并降低了PrP(c)的水平。在没有抗体的情况下再培养14天后，细胞中仍无PrP(Sc)，而PrP(c)水平得以恢复。我们的结果证明了层粘连蛋白受体（LRP/LR）在培养细胞中PrP(Sc)传播的必要性，并表明LRP/LR特异性抗体可作为治疗传染性海绵状脑病的有力治疗工具。
