Immunohistochemical Detection of Disease- Associated Prion Protein in the Peripheral Nervous System in Experimental H-Type Bovine Spongiform Encephalopathy
Immunohistochemical Detection of Disease- Associated Prion Protein in the
Peripheral Nervous System in Experimental H-Type Bovine Spongiform
Encephalopathy
H. Okada1, Y. Iwamaru1, T. Yokoyama1, and S. Mohri1
Abstract
H-type bovine spongiform encephalopathy (BSE) has been identified in aged
cattle in Europe and North America. To determine the localization of
disease-associated prion protein (PrPSc) in the peripheral nerve tissues of
cattle affected with H-type BSE, we employed highly sensitive
immunohistochemical and immunofluorescence techniques with the tyramide signal
amplification (TSA) system. PrPSc deposition was detected in the inferior
ganglia, sympathetic nerve trunk, vagus nerve, spinal nerves, cauda equina, and
adrenal medulla, using this system. Notably, granular PrPSc deposits were
present mainly in the Schwann cells and fibroblastlike cells and occasionally
along certain nerve fibers at the surface of the axons. In the adrenal gland,
PrPSc immunolabeling was observed within the sympathetic nerve fibers and nerve
endings in the adrenal medulla. Although our results were limited to only 3
experimental cases, these results suggest that the TSA system, a highly
sensitive immunohistochemical procedure, may help in elucidating the peripheral
pathogenesis of H-type BSE.
Keywords
atypical bovine spongiform encephalopathy, H-type, prion, tyramide
amplification, peripheral nervous system
snip...
Neuropil, but not neuronal, vacuolation was obvious in all brain areas,
especially in the nuclei of the thalamus and brainstem. Immunolabeled PrPSc was
widely distributed throughout the brain and spinal cord (Fig. 1). Eight patterns
of PrPSc immunolabeling including intraneuronal, perineuronal, intraglial,
linear, fine particulate, coarse granular, stellate, and plaques were identified
in the brain. The most conspicuous pattern of PrPSc was that of fine particulate
and coarse granular deposits in the cerebral and cerebellar cortices and the
nuclei of the thalamus, brainstem, and cerebellum. Stellate-type PrPSc
deposition was detected in the cerebral and cerebellar cortices, basal ganglia,
thalamus, hypothalamus, and hippocampus. Intraglial-type PrPSc deposition was
conspicuous throughout the white matter of the brain and spinal cord. Spherical
PrPSc plaques were sparsely located in the thalamus, basal ganglia, midbrain,
pons, deeper layers of the cerebral cortices, and subcortical white matter.
Immunolabeled PrPSc was observed in the retina, neurohypophysis, and optic
nerve. In the trigeminal and dorsal root ganglia, PrPSc was mainly found
accumulated in both ganglionic and satellite cells, using the conventional
polymer immunodetection method as described previously.9 In addition to the
extracerebral tissues described above, positive PrPSc immunolabeling was
detected in the adrenal gland, cauda equina, cervical spinal nerves, facial
nerve, hypoglossal nerve, vagus nerve, sciatic nerve, and ganglia, such as the
inferior ganglion of the vagus nerve, superior cervical ganglion, stellate
ganglion, ganglia of the sympathetic trunk, and celiac and mesenteric ganglion
complex, with the use of the TSA-biotin system. In these ganglia, not all
neurons of ganglia exhibited intracytoplasmic labeling, but labeling was evident
in the Schwann cells or fibroblast-like cells (Figs. 2, 3). In the cauda equina,
dorsal roots of the cervical spinal nerves, facial nerve, hypoglossal nerve,
vagus nerve, and sciatic nerve, granular PrPSc aggregate was mainly located in
Schwann cells. In the adrenal gland, the PrPSc signal was evident at the
intercellular fine processes of the nerve endings between the chromaffin cells
of the adrenal medulla (Fig. 4). Immunolabeled PrPSc was not detected in the
enteric nervous system such as the myenteric and submucosal plexi of the
digestive tract. No specific immunolabeling was detectable in the lymphoid
tissues with the TSA-biotin system.
Using the dual or triple immunofluorescence technique, localization of
PrPSc was visible in the ganglionic and satellite cells of the ganglia in the
merged images (Fig. 5). In addition, the merged image showed that PrPSc granules
were rarely located at the periphery of axons or within the axons adjacent to
the ganglia (Fig. 6). Moreover, granular PrPSc was observed outside
S100-positive cells (Fig. 7). In the cauda equina and spinal nerves, PrPSc
coexisted mostly in the periphery of Schwann cells labeled with MBP and S100
(Fig. 8). No PrPSc immunolabeling was detected in the compact layers of the
myelin sheath. In addition, no positive signal was detected in the sections from
uninfected controls incubated with mAb F99/97.6.1 by both TSA-biotin and
TSA-fluorescence methods, and no background immunostaining was observed in any
sections of H-type infected animals by both methods when non-immune mouse and
rabbit IgG, or PBS, were applied to the sections instead of the primary
PrP-antibody (Figs. 9, 10).
snip...see full text ;
snip...
Unfortunately, a detailed and all-encompassing analysis of neuropathology
and topographical distribution of immunolabeled PrPSc in H-type BSE-affected
cattle could not be performed, since only the obex region is routinely sampled
for BSE surveillance testing and the remaining brain as well as the carcasses
are not available in most countries [3,10,12,13,24-27]. Recently, clinical signs
and biochemical properties of experimental German H-type BSE cases have been
reported [20]. The primary objective of this study was to investigate the
transmissibility of H-type BSE, using a field isolate detected in the active
surveillance program in Canada [12]. The secondary objective was to extend the
knowledge of the topographical distribution and deposition patterns of
immunolabeled PrPSc in H-type BSE.
snip...
In addition, the present data will support risk assessments in some
peripheral tissues derived from cattle affected with H-type BSE.
Friday, March 09, 2012
Experimental H-type and L-type bovine spongiform encephalopathy in cattle:
observation of two clinical syndromes and diagnostic challenges Research article
Thursday, June 23, 2011
Experimental H-type bovine spongiform encephalopathy characterized by
plaques and glial- and stellate-type prion protein deposits
P.4.23 Transmission of atypical BSE in humanized mouse models
Liuting Qing1, Wenquan Zou1, Cristina Casalone2, Martin Groschup3, Miroslaw
Polak4, Maria Caramelli2, Pierluigi Gambetti1, Juergen Richt5, Qingzhong Kong1
1Case Western Reserve University, USA; 2Instituto Zooprofilattico Sperimentale,
Italy; 3Friedrich-Loeffler-Institut, Germany; 4National Veterinary Research
Institute, Poland; 5Kansas State University (Previously at USDA National Animal
Disease Center), USA
Background: Classical BSE is a world-wide prion disease in cattle, and the
classical BSE strain (BSE-C) has led to over 200 cases of clinical human
infection (variant CJD). Atypical BSE cases have been discovered in three
continents since 2004; they include the L-type (also named BASE), the H-type,
and the first reported case of naturally occurring BSE with mutated bovine PRNP
(termed BSE-M). The public health risks posed by atypical BSE were largely
undefined.
Objectives: To investigate these atypical BSE types in terms of their
transmissibility and phenotypes in humanized mice.
Methods: Transgenic mice expressing human PrP were inoculated with several
classical (C-type) and atypical (L-, H-, or Mtype) BSE isolates, and the
transmission rate, incubation time, characteristics and distribution of PrPSc,
symptoms, and histopathology were or will be examined and compared.
Results: Sixty percent of BASE-inoculated humanized mice became infected
with minimal spongiosis and an average incubation time of 20-22 months, whereas
only one of the C-type BSE-inoculated mice developed prion disease after more
than 2 years. Protease-resistant PrPSc in BASE-infected humanized Tg mouse
brains was biochemically different from bovine BASE or sCJD. PrPSc was also
detected in the spleen of 22% of BASE-infected humanized mice, but not in those
infected with sCJD. Secondary transmission of BASE in the humanized mice led to
a small reduction in incubation time.
*** The atypical BSE-H strain is also transmissible with distinct
phenotypes in the humanized mice, but no BSE-M transmission has been observed so
far.
Discussion: Our results demonstrate that BASE is more virulent than
classical BSE, has a lymphotropic phenotype, and displays a modest transmission
barrier in our humanized mice. BSE-H is also transmissible in our humanized Tg
mice. The possibility of more than two atypical BSE strains will be discussed.
Supported by NINDS NS052319, NIA AG14359, and NIH AI 77774.
P26 TRANSMISSION OF ATYPICAL BOVINE SPONGIFORM ENCEPHALOPATHY (BSE) IN
HUMANIZED MOUSE MODELS
Liuting Qing1, Fusong Chen1, Michael Payne1, Wenquan Zou1, Cristina
Casalone2, Martin Groschup3, Miroslaw Polak4, Maria Caramelli2, Pierluigi
Gambetti1, Juergen Richt5*, and Qingzhong Kong1 1Department of Pathology, Case
Western Reserve University, Cleveland, OH 44106, USA; 2CEA, Istituto
Zooprofilattico Sperimentale, Italy; 3Friedrich-Loeffler-Institut, Germany;
4National Veterinary Research Institute, Poland; 5Kansas State University,
Diagnostic Medicine/Pathobiology Department, Manhattan, KS 66506, USA. *Previous
address: USDA National Animal Disease Center, Ames, IA 50010, USA
Classical BSE is a world-wide prion disease in cattle, and the classical
BSE strain (BSE-C) has led to over 200 cases of clinical human infection
(variant CJD). Two atypical BSE strains, BSE-L (also named BASE) and BSE-H, have
been discovered in three continents since 2004. The first case of naturally
occurring BSE with mutated bovine PrP gene (termed BSE-M) was also found in 2006
in the USA. The transmissibility and phenotypes of these atypical BSE
strains/isolates in humans were unknown. We have inoculated humanized transgenic
mice with classical and atypical BSE strains (BSE-C, BSE-L, BSE-H) and the BSE-M
isolate. We have found that the atypical BSE-L strain is much more virulent than
the classical BSE-C.
*** The atypical BSE-H strain is also transmissible in the humanized
transgenic mice with distinct phenotype, but no transmission has been observed
for the BSE-M isolate so far.
III International Symposium on THE NEW PRION BIOLOGY: BASIC SCIENCE,
DIAGNOSIS AND THERAPY 2 - 4 APRIL 2009, VENEZIA (ITALY)
I ask Professor Kong ;
Thursday, December 04, 2008 3:37 PM
Subject: RE: re--Chronic Wating Disease (CWD) and Bovine Spongiform
Encephalopathies (BSE): Public Health Risk Assessment
''IS the h-BSE more virulent than typical BSE as well, or the same as cBSE,
or less virulent than cBSE? just curious.....''
Professor Kong reply ;
.....snip
''As to the H-BSE, we do not have sufficient data to say one way or
another, but we have found that H-BSE can infect humans. I hope we could publish
these data once the study is complete. Thanks for your interest.'' Best regards,
Qingzhong Kong, PhD Associate Professor Department of Pathology Case Western
Reserve University Cleveland, OH 44106 USA
END...TSS
Thursday, December 04, 2008 2:37 PM
"we have found that H-BSE can infect humans." personal communication with
Professor Kong. ...TSS
BSE-H is also transmissible in our humanized Tg mice.
The possibility of more than two atypical BSE strains will be discussed.
Supported by NINDS NS052319, NIA AG14359, and NIH AI 77774.
http://www.neuroprion.org/resources/pdf_docs/conferences/prion2009/prion2009_bookofabstracts.pdf
http://transmissiblespongiformencephalopathy.blogspot.com/2011/06/experimental-h-type-bovine-spongiform.html
SNIP...
Friday, May 11, 2012
Experimental H-type bovine spongiform encephalopathy characterized by
plaques and glial- and stellate-type prion protein deposits
Thursday, June 21, 2012
Clinical and Pathologic Features of H-Type Bovine Spongiform Encephalopathy
Associated with E211K Prion Protein Polymorphism
Tuesday, November 02, 2010
IN CONFIDENCE
The information contained herein should not be disseminated further except
on the basis of "NEED TO KNOW".
BSE - ATYPICAL LESION DISTRIBUTION (RBSE 92-21367) statutory (obex only)
diagnostic criteria CVL 1992
2009 UPDATE ON ALABAMA AND TEXAS MAD COWS 2005 and 2006
let's take a closer look at this new prionpathy or prionopathy, and then
let's look at the g-h-BSEalabama mad cow.
This new prionopathy in humans? the genetic makeup is IDENTICAL to the
g-h-BSEalabama mad cow, the only _documented_ mad cow in the world to date like
this, ......wait, it get's better. this new prionpathy is killing young and old
humans, with LONG DURATION from onset of symptoms to death, and the symptoms are
very similar to nvCJD victims, OH, and the plaques are very similar in some
cases too, bbbut, it's not related to the g-h-BSEalabama cow, WAIT NOW, it gets
even better, the new human prionpathy that they claim is a genetic TSE, has no
relation to any gene mutation in that family. daaa, ya think it could be related
to that mad cow with the same genetic make-up ??? there were literally tons and
tons of banned mad cow protein in Alabama in commerce, and none of it
transmitted to cows, and the cows to humans there from ??? r i g h t $$$
ALABAMA MAD COW g-h-BSEalabama
In this study, we identified a novel mutation in the bovine prion protein
gene (Prnp), called E211K, of a confirmed BSE positive cow from Alabama, United
States of America. This mutation is identical to the E200K pathogenic mutation
found in humans with a genetic form of CJD. This finding represents the first
report of a confirmed case of BSE with a potential pathogenic mutation within
the bovine Prnp gene. We hypothesize that the bovine Prnp E211K mutation most
likely has caused BSE in "the approximately 10-year-old cow" carrying the E221K
mutation.
her healthy calf also carried the mutation (J. A. Richt and S. M. Hall PLoS
Pathog. 4, e1000156; 2008).
This raises the possibility that the disease could occasionally be genetic
in origin. Indeed, the report of the UK BSE Inquiry in 2000 suggested that the
UK epidemic had most likely originated from such a mutation and argued against
the scrapierelated assumption. Such rare potential pathogenic PRNP mutations
could occur in countries at present considered to be free of BSE, such as
Australia and New Zealand. So it is important to maintain strict surveillance
for BSE in cattle, with rigorous enforcement of the ruminant feed ban (many
countries still feed ruminant proteins to pigs). Removal of specified risk
material, such as brain and spinal cord, from cattle at slaughter prevents
infected material from entering the human food chain. Routine genetic screening
of cattle for PRNP mutations, which is now available, could provide additional
data on the risk to the public. Because the point mutation identified in the
Alabama animals is identical to that responsible for the commonest type of
familial (genetic) CJD in humans, it is possible that the resulting infective
prion protein might cross the bovine–human species barrier more easily. Patients
with vCJD continue to be identified. The fact that this is happening less often
should not lead to relaxation of the controls necessary to prevent future
outbreaks. Malcolm A. Ferguson-Smith Cambridge University Department of
Veterinary Medicine, Madingley Road, Cambridge CB3 0ES, UK e-mail:
maf12@cam.ac.uk Jürgen A. Richt College of Veterinary Medicine, Kansas State
University, K224B Mosier Hall, Manhattan, Kansas 66506-5601, USA
NATURE|Vol 457|26 February 2009
Saturday, August 14, 2010
BSE Case Associated with Prion Protein Gene Mutation (g-h-BSEalabama) and
VPSPr PRIONPATHY (see mad cow feed in COMMERCE IN ALABAMA...TSS)
Tuesday, November 6, 2012
***Transmission of New Bovine Prion to Mice, Atypical Scrapie, BSE, and
Sporadic CJD, November-December 2012 update
Comments on technical aspects of the risk assessment were then submitted to
FSIS.
Comments were received from Food and Water Watch, Food Animal Concerns
Trust (FACT), Farm Sanctuary, R-CALF USA, Linda A Detwiler, and Terry S.
Singeltary.
This document provides itemized replies to the public comments received on
the 2005 updated Harvard BSE risk assessment. Please bear the following points
in mind:
Owens, Julie
From: Terry S. Singeltary Sr. [flounder9@verizon.net]
Sent: Monday, July 24, 2006 1:09 PM
To: FSIS RegulationsComments
Subject: [Docket No. FSIS-2006-0011] FSIS Harvard Risk Assessment of Bovine
Spongiform Encephalopathy (BSE) Page 1 of 98
FSIS, USDA, REPLY TO SINGELTARY
U.S.A. 50 STATE BSE MAD COW CONFERENCE CALL Jan. 9, 2001
2012 atypical L-type BSE BASE California reports
SUMMARY REPORT CALIFORNIA BOVINE SPONGIFORM ENCEPHALOPATHY CASE
INVESTIGATION JULY 2012
Summary Report BSE 2012
Executive Summary
Saturday, August 4, 2012
Update from APHIS Regarding Release of the Final Report on the BSE
Epidemiological Investigation
Saturday, August 4, 2012
*** Final Feed Investigation Summary - California BSE Case - July 2012
Saturday, December 15, 2012
Bovine spongiform encephalopathy: the effect of oral exposure dose on
attack rate and incubation period in cattle -- an update 5 December 2012
Thursday, December 20, 2012
OIE GROUP RECOMMENDS THAT SCRAPE PRION DISEASE BE DELISTED AND SAME OLD BSe
WITH BOVINE MAD COW DISEASE
Tuesday, December 25, 2012
A Quantitative Assessment of the Amount of Prion Diverted to Category 1
Materials and Wastewater During Processing
2011 Monday, September 26, 2011
L-BSE BASE prion and atypical sporadic CJD
Tuesday, June 26, 2012
Creutzfeldt Jakob Disease Human TSE report update North America, Canada,
Mexico, and USDA PRION UNIT as of May 18, 2012
type determination pending Creutzfeldt Jakob Disease (tdpCJD), is on the
rise in Canada and the USA
Sunday, December 2, 2012
CANADA 19 cases of mad cow disease SCENARIO 4: ‘WE HAD OUR CHANCE AND WE
BLEW IT’
Monday, July 23, 2012
The National Prion Disease Pathology Surveillance Center July 2012
Tuesday, December 25, 2012
CREUTZFELDT JAKOB TSE PRION DISEASE HUMANS END OF YEAR REVIEW DECEMBER 25,
2012
Monday, December 31, 2012
Creutzfeldt Jakob Disease and Human TSE Prion Disease in Washington State,
2006–2011-2012
TSS
posted by Terry S. Singeltary Sr. at
1:10 PM
0 Comments:
Post a Comment
Subscribe to Post Comments [Atom]
<< Home