Comparison of incubation periods in wild-type mice.

<p>Incubation period plot comparison of vCJD (transfusion) case versus transmissions in wild-type mice of vCJD (BSE) from three sources. (Data shows mean incubation period±standard error of the mean. Open circles RIII line and open triangles VM line.)</p

Detection of abnormal PrP in the mouse brain.

<p>Immunocytochemical detection of abnormal PrP deposition in hippocampus and thalamus (lateral posterior nucleus) of HuMM transgenic (with additional 40× magnification of florid plaque structure, see box lower left) and VM wild-type mice following inoculation with vCJD (BSE) and vCJD (transfusion) material. (Scale bar 200 µm, anti-PrP antibody 6H4)</p

Vacuolation scoring in the mouse brain.

<p>Lesion profile comparison of vCJD (transfusion) case versus vCJD (BSE) transmissions to identify similarities in vacuolar pathology levels and regional distribution in mouse brains. (mean score ±SEM; dashed line - vCJD (transfusion) case; solid lines – 3x vCJD (BSE) cases for wild-type mice (diamonds – vCJD(BSE) A; squares – vCJD(BSE) B; triangles – vCJD(BSE) C) and published vCJD (BSE) for HuMM transgenic; G1–G9 grey matter scoring regions; W1–W3 white matter scoring regions)</p

Clinical and pathological assessment of wild-type mice.

<p>Wild-type mouse lines RIII and VM, inoculated with vCJD(BSE) and vCJD(transfusion) were assessed clinically and pathologically for signs of TSE and mean incubation periods calculated.</p>a<p>The group of 24 was reduced due to unavailability of some brain material for analysis.</p

PrP^Sc typing by Western blot.

<p>Brain homogenates from HuMM mice inoculated with both vCJD (BSE) and vCJD (transfusion) show similar mobility and glycosylation profile (type 2B) as material from vCJD patients. (T2B: control vCJD material; antibody: 6H4)</p

Lesion Profiles in Voles and Mice following Transmission of sCJD and gCJD

<div><p>Brain-scoring positions are medulla (1), cerebellum (2), superior colliculus (3), hypothalamus (4), thalamus (5), hippocampus (6), septum (7), retrosplenial and adjacent motor cortex (8), and cingulate and adjacent motor cortex (9).</p><p>(A) Lesion profiles in voles infected with MM1 sCJD (denoted by open squares), MV1 sCJD (denoted by open circles), V210I gCJD (denoted by open diamonds), and E200K gCJD (denoted by open upright triangles).</p><p>(B) Lesion profiles in voles infected with MM2 sCJD (denoted by open squares) and the mouse-passaged scrapie strain ME7 (denoted by open circles).</p><p>(C) Lesion profiles in voles infected with human sCJD (denoted by open squares and open circles) and mouse-passaged sCJD (denoted by open diamonds and open upright triangles), MM1 sCJD (denoted by open squares and open diamonds), and MV1 sCJD (denoted by open circles and open upright triangles).</p><p>(D) Lesion profiles in C3H mice infected with MM1 sCJD (denoted by open circles), MV1 sCJD (denoted by open upright triangles), and V210I gCJD (denoted by open diamonds).</p></div

Regional Distribution of Protease-Resistant PrP^Sc in Voles following Transmission of sCJD and gCJD

<p>PET blots of coronal sections of the forebrain (telencephalon in [A] and diencephalon in [B]), midbrain (C), and hindbrain (D) in voles infected with MM2 sCJD, MM1 sCJD, MV1 sCJD, V210I gCJD, and E200K gCJD. At the lower part of the figure, the labeled coronal sections of a negative control brain are shown. NC, neocortex; Sp, septum; St, striatum; Hp, hippocampus; Th, thalamus; Hy, hypothalamus; SC, superior colliculus; GN, geniculate nuclei; SN, substantia nigra; Cb, cerebellum; MO, medulla oblongata.</p

Determination of the Molecular Type of PrP^Sc Produced in Voles following Transmission of sCJD and gCJD

<div><p>(A) Immunoblot of proteinase K–resistant PrP^Sc from sCJD and gCJD subtypes in human patients and after first passage in voles. PrP^Sc produced in voles after primary transmission of mouse-passaged MM1 sCJD and MV1 sCJD, and of the mouse-adapted scrapie strain ME7 is also shown. The identity of the brain sample is designated above each lane.</p><p>(B) Comparison of proteinase K–resistant PrP^Sc produced in voles following first and second passages of MV1 sCJD, MM2 sCJD, E200K gCJD, and V210I gCJD. PrP^Sc produced in voles after first passage of the mouse-adapted scrapie strain ME7 is also shown. The identity of the brain sample is designated above each lane.</p><p>(C) Scatter-graph of proportions of di-glycosylated and mono-glycosylated PrP^Sc in human patients (denoted by filled circles, filled inverted triangles, filled diamonds, filled squares, and filled upright triangles) and voles (denoted by open circles, open inverted triangles, open diamonds, open squares, open upright triangles, and asterisks), with MM1 sCJD (denoted by filled squares and open squares), MV1 sCJD (denoted by filled diamonds and open diamonds), MM2 sCJD (denoted by filled circles and open circles), V210I gCJD (denoted by filled inverted triangles and open inverted triangles), E200K gCJD (denoted by filled upright triangles and open upright triangles), and ME7 (denoted by asterisks).</p><p>(D) Comparison of proteinase K–resistant PrP^Sc produced in voles following inoculation with MM1 sCJD, MM2 sCJD, and the mouse-adapted scrapie strain ME7. Strep-tagged molecular markers (25 kDa and 20 kDa) are shown. The identity of the brain samples is designated above each lane.</p></div

Alignment of Human, Vole, and Mouse Prion Protein–Amino Acid Sequences

<p>The sequence numbers of the human <i>(Homo sapiens)</i> amino acids are indicated and refer to the residue under the final digit. In the vole <i>(C. glareolus)</i> and the mouse <i>(Mus musculus)</i> sequences, identical residues to the human are indicated as dots.</p

Comparison of Transmission Barriers of Human, Sheep, Mouse, and Hamster Prions following Transmission to Voles

<div><p>In (A–C), inocula are grouped according to the sequence homology of mature PrP between the donor species and voles, from the least to the most divergent (golden hamster, 2.9% divergence; C57BL mouse strain, 2.9% divergence; VM mouse strain, 3.4% divergence; human, 7.1% divergence; and sheep, 7.1% divergence). In (D), inocula are ranked according to the magnitude of the transmission barrier and are grouped according to the original prion source.</p><p>(A) Survival times following first passage in voles of hamster-passaged scrapie strains 263K and 139H, C57BL mice-passaged BSE strain 301C, C57BL mice-passaged scrapie strains ME7 and 139A, VM mice-passaged BSE strain 301V, VM mice-passaged scrapie strain 22A, human sCJD (MM1, MV1, and MM2 subtypes) and gCJD (E200K and V210I subtypes), sheep-passaged BSE, and sheep natural scrapie isolates from the United Kingdom (SSUK) and Italy (SSIT); error bars represent standard deviation.</p><p>(B) Survival times following second passage in voles of the same transmissions as in (A); error bars represent standard deviation.</p><p>(C and D) The magnitude of each transmission barrier is calculated as the ratio of mean survival times observed following first and second passage (a value of 1 denotes an absence of transmission barrier).</p></div