section 9 is highlighted. 3. Focus on Section 9. How can researchers and conservationists test for the presence of the prion associated with CWD? Why is testing difficult? What is the treatment? Do your answers to these questions make sense given your knowledge of protein structure and folding? for CWD transmission to humans, one report rectal mucos, and brain in CWD positive elk. indicates nonhuman primate susceptibility to alhough one of seven elk with early CWD CWD. Intracerebral inoculation of squirrel was tonsil positive, but rectal mucosa negamonkeys (Saimini sciureus) demonstrated a tive [82]. positive CWD transmission [49]. Among non- PrP detection in CWD-infected deer human primates, however, the Pmp sequence: blood has been a challenge to develop as an anof the new world monkeys are the most dis- temortem diagnostic tool. Prptic was recently tant from humans [72], and therefore may not reported as detectable in deer blood using an indicate that human prion protein conversion antibody ELISA with signal amplification of would be induced by CWD PrP S c . antibody conjugated DNA catalyzed by T7 RNA polymerase [15]. 1. CWD DIAGNOSTICS Protein misfolding cyclic amplification (PMCA) can be used to amplify PrPir from Clinical kigns of CWD in deer and elk CWD-infected deer, and should be valuable are unspecific and-subtle in cariy disease and for Prpes detection from tissues and body flucommonly include weight loss and behav- ids containing low levels of infectivity. for foral changes such as isolation from the herd improved understanding of CWD pathogeneand depression. Other signs may include hy: improved understanding of CWD palso to detect animals in early stages persalivation, polydipsia/polyuria, ataxia, and of infection [43]. Atarashi et al. [3] developed occisionally increased regurgitation and/or of infection [43]. Atarashi ct al. [3/ developed esophageal distension. Therefore sensitive, nique whereby recombinant, natively folded specific, and rapid ante-mortem CWD ascays Pique whereby recombinant, natively folded are eritical for accurafe diagnosis. Removal PrP monomers is used to amp 50 ag of PrPs was detectable. Therefore of infected animals from a berd, pairticularly methods for sensitive detection of Prpe are in US National Parks where culling of only subatantially improvingl known infected individuals is often the preferred method of management ( M . Wild, personal communication). To survey and man- 10. TOOLS FOR CWD RESEARCH: CELL. age CWD in Rocky Mountain National Park IINES AND CERVID PrP EXPRESSING and adjacent Estes Park in Colondo.male TRANSGENIC MICE deer were anesthetized, radiocollared, and ton- Until recently, few techniques were availsil biopsies collected and tected (M. Wild, able to detect CWD prion infectivity, or to personal communication: [941) Any prion: test compounds to disrupt prion conversion. A infected animals were then located by ra- CWD-susceptible cell line derived from cervid diotelemetry and euthani.

1. section 9 is highlighted.
3. Focus on Section 9. How can researchers and conservationists test for the presence of the
prion associated with CWD? Why is testing difficult? What is the treatment? Do your answers to
these questions make sense given your knowledge of protein structure and folding? for CWD
transmission to humans, one report rectal mucos, and brain in CWD positive elk. indicates
nonhuman primate susceptibility to alhough one of seven elk with early CWD CWD.
Intracerebral inoculation of squirrel was tonsil positive, but rectal mucosa negamonkeys (Saimini
sciureus) demonstrated a tive [82]. positive CWD transmission [49]. Among non- PrP detection
in CWD-infected deer human primates, however, the Pmp sequence: blood has been a challenge
to develop as an anof the new world monkeys are the most dis- temortem diagnostic tool. Prptic
was recently tant from humans [72], and therefore may not reported as detectable in deer blood
using an indicate that human prion protein conversion antibody ELISA with signal amplification
of would be induced by CWD PrP S c . antibody conjugated DNA catalyzed by T7 RNA
polymerase [15]. 1. CWD DIAGNOSTICS Protein misfolding cyclic amplification (PMCA) can
be used to amplify PrPir from Clinical kigns of CWD in deer and elk CWD-infected deer, and
should be valuable are unspecific and-subtle in cariy disease and for Prpes detection from tissues
and body flucommonly include weight loss and behav- ids containing low levels of infectivity.
for foral changes such as isolation from the herd improved understanding of CWD pathogeneand
depression. Other signs may include hy: improved understanding of CWD palso to detect
animals in early stages persalivation, polydipsia/polyuria, ataxia, and of infection [43]. Atarashi
et al. [3] developed occisionally increased regurgitation and/or of infection [43]. Atarashi ct al.
[3/ developed esophageal distension. Therefore sensitive, nique whereby recombinant, natively
folded specific, and rapid ante-mortem CWD ascays Pique whereby recombinant, natively folded
are eritical for accurafe diagnosis. Removal PrP monomers is used to amp 50 ag of PrPs was
detectable. Therefore of infected animals from a berd, pairticularly methods for sensitive
detection of Prpe are in US National Parks where culling of only subatantially improvingl known
infected individuals is often the preferred method of management ( M . Wild, personal
communication). To survey and man- 10. TOOLS FOR CWD RESEARCH: CELL. age CWD in
Rocky Mountain National Park IINES AND CERVID PrP EXPRESSING and adjacent Estes
Park in Colondo.male TRANSGENIC MICE deer were anesthetized, radiocollared, and ton-
Until recently, few techniques were availsil biopsies collected and tected (M. Wild, able to detect
CWD prion infectivity, or to personal communication: [941) Any prion: test compounds to
disrupt prion conversion. A infected animals were then located by ra- CWD-susceptible cell line
derived from cervid diotelemetry and euthanized. Therefore, in- brain fibroblasts has been used
to screen intensive CWD management can be costly and hibitors of CWD infection, for example,
penlabor intensive. tosan polysulfate [69]. This CWD specific as- Rectal biopsies have been
evaluated as an say may identify compounds that inhibit CWD aliernative to tonsil biopsies for
large scale propagation. surveillance of free-ranging or captive deer Browning et al. developed
the first transPrPr was readily detected by IHC in ree- genic moose expressing cervid PrP [10],
which tal lymphoid follicles from experimentally in- was subsequently utilized as a bioassay to
feeted deer. where 19 of 19 deer were posifive detect CWD prions in muscle. [2]. Several by one
year after oral inoculation. In maturaty. transgenic mice expressing cervid PrP have infected
mule deer. 45 of 50 subelinical and since been produced using different constructs ferminal
CWD-infected deer, as determinedby and sequences [ 10 , 41 , 44 , 52 , 84 ] (detailed in
immunohistochemistry on tonsil or retrophas. Tab. D). The cervid PrP-expressing transgenic
ryngeal lymph nodes. Were also positive by mice provide an important tool for investigareetal

2. biopsy [95]. Rectal biopsies to diagnose tions of CWD and other TSE, particularly for CWD in
elk may also be suitable, as one study cross-species transmission studies. In a study showed a
strong correlation between tonsil. by Trifilo et al. cervid PeP expressing mice