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Acquired transmissibility of sheep-passaged L-type bovine spongiform encephalopathy prion to wild-type mice.

Okada H, Masujin K, Miyazawa K, Yokoyama T - Vet. Res. (2015)

Bottom Line: Disease-associated prion protein (PrP(Sc)) was detected in the brain and/or lymphoid tissues during the lifespan of mice that were asymptomatic subclinical carriers, indicating that wild-type mice were susceptible to sheep-passaged L-BSE.The morphological characteristics of the PrP(Sc) of sheep-passaged L-BSE included florid plaques that were distributed mainly in the cerebral cortex and hippocampus of subsequent passaged mice.The PrP(Sc) glycoform profiles of wild-type mice infected with sheep-passaged L-BSE were similar to those of the original isolate.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Animal Health, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan. okadahi@affrc.go.jp.

ABSTRACT
L-type bovine spongiform encephalopathy (L-BSE) is an atypical form of BSE that is transmissible to cattle and several lines of prion protein (PrP) transgenic mice, but not to wild-type mice. In this study, we examined the transmissibility of sheep-passaged L-BSE prions to wild-type mice. Disease-associated prion protein (PrP(Sc)) was detected in the brain and/or lymphoid tissues during the lifespan of mice that were asymptomatic subclinical carriers, indicating that wild-type mice were susceptible to sheep-passaged L-BSE. The morphological characteristics of the PrP(Sc) of sheep-passaged L-BSE included florid plaques that were distributed mainly in the cerebral cortex and hippocampus of subsequent passaged mice. The PrP(Sc) glycoform profiles of wild-type mice infected with sheep-passaged L-BSE were similar to those of the original isolate. The data indicate that sheep-passaged L-BSE has an altered host range and acquired transmissibility to wild-type mice.

No MeSH data available.


Related in: MedlinePlus

Western blot analysis of proteinase-K resistant PrPSc analyzed using monoclonal antibody T2.A PrPSc in the brain (Br) and spleen (Sp) of wild-type mice inoculated with sheep-passaged L-BSE at the first and second passage. All samples were digested with 50 μg/mL of proteinase-K at 37 °C for 1 h. Lanes from left to right were loaded with 0.625, 5, 0.0125, and 0.36 mg tissue equivalent, respectively. The molecular markers are shown on the left (kDa). B PrPSc in the brain of C-BSE- and L-BSE-affected cattle and mice. Lane 1: C-BSE affected cattle, Lane 2: C-BSE affected ICR mouse, Lane 3: L-BSE affected cattle, Lane 4: L-BSE affected sheep, and Lane 5: sheep-passaged L-BSE affected ICR mouse at second passage. Lanes 1, 3, and 4, and Lanes 2 and 5 were loaded with 1.25 and 0.125 mg tissue equivalent, respectively. C Quantification of the relative amounts of the di-, mono-, and unglycosylated forms of PrPSc from the brain. The column numbers are as listed in (B). Bar diagram indicates the diglycosylated form (black), monoglycosylated form (gray), and unglycosylated form (white). Data are expressed as mean ± standard deviation of triplicate experiments.
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Fig3: Western blot analysis of proteinase-K resistant PrPSc analyzed using monoclonal antibody T2.A PrPSc in the brain (Br) and spleen (Sp) of wild-type mice inoculated with sheep-passaged L-BSE at the first and second passage. All samples were digested with 50 μg/mL of proteinase-K at 37 °C for 1 h. Lanes from left to right were loaded with 0.625, 5, 0.0125, and 0.36 mg tissue equivalent, respectively. The molecular markers are shown on the left (kDa). B PrPSc in the brain of C-BSE- and L-BSE-affected cattle and mice. Lane 1: C-BSE affected cattle, Lane 2: C-BSE affected ICR mouse, Lane 3: L-BSE affected cattle, Lane 4: L-BSE affected sheep, and Lane 5: sheep-passaged L-BSE affected ICR mouse at second passage. Lanes 1, 3, and 4, and Lanes 2 and 5 were loaded with 1.25 and 0.125 mg tissue equivalent, respectively. C Quantification of the relative amounts of the di-, mono-, and unglycosylated forms of PrPSc from the brain. The column numbers are as listed in (B). Bar diagram indicates the diglycosylated form (black), monoglycosylated form (gray), and unglycosylated form (white). Data are expressed as mean ± standard deviation of triplicate experiments.

Mentions: Conventional WB features of PK-resistant PrPSc, such as the electrophoretic mobility and the relative proportions and pattern of the glycoforms obtained using the mAbs T2 and SAF84, were similar between the first and the second passage in the brain and spleen of wild-type mice inoculated with L-BSE/sheep (Figure 3A). The molecular mass of the unglycosylated form was ~18 kDa in cattle, sheep, and wild-type mice affected with L-BSE. Interestingly, the three bands in samples from wild-type mice represented higher molecular mass forms compared with those of cattle and sheep affected with L-BSE (Figure 3B). Of note, the PrPSc glycoprofile of L-BSE/sheep-affected wild-type mice resembled those of L-BSE-affected cattle and sheep, but was distinct from those of C-BSE-affected cattle and wild-type mice (Figure 3C).Figure 3


Acquired transmissibility of sheep-passaged L-type bovine spongiform encephalopathy prion to wild-type mice.

Okada H, Masujin K, Miyazawa K, Yokoyama T - Vet. Res. (2015)

Western blot analysis of proteinase-K resistant PrPSc analyzed using monoclonal antibody T2.A PrPSc in the brain (Br) and spleen (Sp) of wild-type mice inoculated with sheep-passaged L-BSE at the first and second passage. All samples were digested with 50 μg/mL of proteinase-K at 37 °C for 1 h. Lanes from left to right were loaded with 0.625, 5, 0.0125, and 0.36 mg tissue equivalent, respectively. The molecular markers are shown on the left (kDa). B PrPSc in the brain of C-BSE- and L-BSE-affected cattle and mice. Lane 1: C-BSE affected cattle, Lane 2: C-BSE affected ICR mouse, Lane 3: L-BSE affected cattle, Lane 4: L-BSE affected sheep, and Lane 5: sheep-passaged L-BSE affected ICR mouse at second passage. Lanes 1, 3, and 4, and Lanes 2 and 5 were loaded with 1.25 and 0.125 mg tissue equivalent, respectively. C Quantification of the relative amounts of the di-, mono-, and unglycosylated forms of PrPSc from the brain. The column numbers are as listed in (B). Bar diagram indicates the diglycosylated form (black), monoglycosylated form (gray), and unglycosylated form (white). Data are expressed as mean ± standard deviation of triplicate experiments.
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Related In: Results  -  Collection

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Fig3: Western blot analysis of proteinase-K resistant PrPSc analyzed using monoclonal antibody T2.A PrPSc in the brain (Br) and spleen (Sp) of wild-type mice inoculated with sheep-passaged L-BSE at the first and second passage. All samples were digested with 50 μg/mL of proteinase-K at 37 °C for 1 h. Lanes from left to right were loaded with 0.625, 5, 0.0125, and 0.36 mg tissue equivalent, respectively. The molecular markers are shown on the left (kDa). B PrPSc in the brain of C-BSE- and L-BSE-affected cattle and mice. Lane 1: C-BSE affected cattle, Lane 2: C-BSE affected ICR mouse, Lane 3: L-BSE affected cattle, Lane 4: L-BSE affected sheep, and Lane 5: sheep-passaged L-BSE affected ICR mouse at second passage. Lanes 1, 3, and 4, and Lanes 2 and 5 were loaded with 1.25 and 0.125 mg tissue equivalent, respectively. C Quantification of the relative amounts of the di-, mono-, and unglycosylated forms of PrPSc from the brain. The column numbers are as listed in (B). Bar diagram indicates the diglycosylated form (black), monoglycosylated form (gray), and unglycosylated form (white). Data are expressed as mean ± standard deviation of triplicate experiments.
Mentions: Conventional WB features of PK-resistant PrPSc, such as the electrophoretic mobility and the relative proportions and pattern of the glycoforms obtained using the mAbs T2 and SAF84, were similar between the first and the second passage in the brain and spleen of wild-type mice inoculated with L-BSE/sheep (Figure 3A). The molecular mass of the unglycosylated form was ~18 kDa in cattle, sheep, and wild-type mice affected with L-BSE. Interestingly, the three bands in samples from wild-type mice represented higher molecular mass forms compared with those of cattle and sheep affected with L-BSE (Figure 3B). Of note, the PrPSc glycoprofile of L-BSE/sheep-affected wild-type mice resembled those of L-BSE-affected cattle and sheep, but was distinct from those of C-BSE-affected cattle and wild-type mice (Figure 3C).Figure 3

Bottom Line: Disease-associated prion protein (PrP(Sc)) was detected in the brain and/or lymphoid tissues during the lifespan of mice that were asymptomatic subclinical carriers, indicating that wild-type mice were susceptible to sheep-passaged L-BSE.The morphological characteristics of the PrP(Sc) of sheep-passaged L-BSE included florid plaques that were distributed mainly in the cerebral cortex and hippocampus of subsequent passaged mice.The PrP(Sc) glycoform profiles of wild-type mice infected with sheep-passaged L-BSE were similar to those of the original isolate.

View Article: PubMed Central - PubMed

Affiliation: National Institute of Animal Health, National Agriculture and Food Research Organization (NARO), Tsukuba, Ibaraki, Japan. okadahi@affrc.go.jp.

ABSTRACT
L-type bovine spongiform encephalopathy (L-BSE) is an atypical form of BSE that is transmissible to cattle and several lines of prion protein (PrP) transgenic mice, but not to wild-type mice. In this study, we examined the transmissibility of sheep-passaged L-BSE prions to wild-type mice. Disease-associated prion protein (PrP(Sc)) was detected in the brain and/or lymphoid tissues during the lifespan of mice that were asymptomatic subclinical carriers, indicating that wild-type mice were susceptible to sheep-passaged L-BSE. The morphological characteristics of the PrP(Sc) of sheep-passaged L-BSE included florid plaques that were distributed mainly in the cerebral cortex and hippocampus of subsequent passaged mice. The PrP(Sc) glycoform profiles of wild-type mice infected with sheep-passaged L-BSE were similar to those of the original isolate. The data indicate that sheep-passaged L-BSE has an altered host range and acquired transmissibility to wild-type mice.

No MeSH data available.


Related in: MedlinePlus