Successful Transmission of Three Mouse-Adapted Scrapie Strains to Murine Neuroblastoma Cell Lines Overexpressing Wild-Type Mouse Prion Protein

Abstract

This new ex vivo transmission model will facilitate research into the mechanism of host-agent interactions, such as the species barrier and strain diversity, and provides a basis for the development of highly susceptible cell lines that could be used in diagnostic and therapeutic approaches to the TSEs.

FK506 reduces abnormal prion protein through the activation of autolysosomal degradation and prolongs survival in prion-infected mice

“…N2a58 and MG20 cells were prepared as described previously. 47,48 Fukuoka-1-infected N2a58 (N2a58/Fukuoka-1) and MG20 cells (MG20/Fukuoka-1) were produced by inoculation with brain homogenates harvested from Fukuoka-1-infected, terminally ill ddY mice. All cell media was supplemented with 10% fetal bovine serum and penicillin-streptomycin (Nacalai, 09367-34).…”

Cell models of prion infection

“…They include N2a [19,115], C-1300 [115] and NIE-115 [86] lines, all of them deriving from the same cell line [114]. Subclones of the N2a cell line with higher permissiveness were subsequently identified [16,41,95] in which subcloning of the infected cultures was no longer necessary to stably maintain high levels of infection. GT1 cells are hypothalamic cells also widely used to multiply mouse prions [5,95,124].…”

“…Subclones of the N2a cell line with higher permissiveness were subsequently identified [16,41,95] in which subcloning of the infected cultures was no longer necessary to stably maintain high levels of infection. GT1 cells are hypothalamic cells also widely used to multiply mouse prions [5,95,124]. Some GT1 subclones may undergo apoptosis following infection [124], a finding not consistently reported.…”

“…In the absence of nucleic acids, strain biological properties are proposed to be encoded by the abnormal PrP [150]. The PrP res banding patterns of strains generated in cultured cells are often, but not always [34], different from those generated in the brain [3,4,26,90,95,146] but return to the original brain pattern upon animal inoculation [3,4]. This confirmed that the cellular context in which multiplication occurs has a determinant effect on the glycoforms and on the size of the abnormal PrP species and that modifications of the PrP res molecular profile are not necessarily associated with changes in their biological properties [133].…”

Caprine PrP variants harboring Asp-146, His-154 and Gln-211 alleles display reduced convertibility upon interaction with pathogenic murine prion protein in scrapie infected cells

“…72 Cells were cultured in OPTIMEM (Gibco) supplemented with 10% (v/v) FBS and 1% (v/v) penicillin/streptomycin, inside a 5% CO 2 atmosphere incubator maintained at 37 C. Media was exchanged every 3 d. Transient transfections were performed on 60% confluent cultures, using the XtermeGene9 reagent (Roche, Cat No 06 365 787 001), according to the manufacturer instructions and a 3:1 ratio of transfection reagent to plasmid DNA. All experiments were conducted on Mycoplasma-free cells.…”