Infection System

Prions & prion diseases

Dr Andrew Thompsett
[email protected]
 Prion Diseases
• Neurodegenerative diseases
• Alternatively known as Transmissible
Spongiform Encephalopathy (TSE)
• Prion (pre-on), proteinaceous
infectious particle
• PrPSc - scrapie form of the prion
protein
• Stanley Prusiner: Nobel prize in
medicine 1997
Prion Diseases
• Affects mammals, including:

Scrapie Bovine Spongiform Chronic Wasting Disease

Encephalopathy (BSE) (CWD)

• In humans
• Creutzfeldt-Jakob disease (CJD)
variant Creutzfeldt-Jakob disease (vCJD)
• Kuru
• Fatal Familiar Insomnia (FFI)
• Gerstmann-Straussler-Scheinke syndrome (GSS)
Signs & symptoms include
• Anxiety & depression
• Ataxia (loss of physical coordination)
• Memory loss and loss of cognition
• Dystonia (muscle spasms)
• Incontinence (bowel & urinary)
• Inevitably fatal: no cures, only treatments to ease
symptoms
• Most people with CJD will die within a year of the
symptoms starting (often from infection)
Disease characteristics

• Neurodegenerative diseases
• Characterised by
• Neuronal death leading to a spongiform (sponge
like) appearance of the brain
• Proliferation of astrocytes & microglia
• Build up of amyloid plaques (protein aggregates)
• Evidence of oxidative stress
 Disease Occurrence
• Very rare
• Prion diseases are unique in medicine in that they
can occur by three mechanisms:
• Sporadic (spontaneous) 85-90% of cases
• symptoms usually develop between the ages of 60 and 65
• 2 deaths / million people / year, 130 deaths in 2023
• Genetic (familial) 10-15% of cases
• symptoms usually develop in early 50s
• 1 / 9-15 million people / year, 4 deaths in 2022
• Acquired (infectious/ transmitted) 2-5% of cases
• V. rare, 1 death from iatrogenic infection in 2023 (0 in 2022)

Data from National CJD surveillance unit:

https://www.cjd.ed.ac.uk/
Prion Pathology
Different forms of disease
affect different parts of the
brain
Linked to strain of the
prion?
 Prion Pathology

Protein plaque - PrPSc Microglia

Spongiform changes
 Amyloid Plaques

•Comprised of protein
•β-sheet rich
•Proteins stack up to
make fibrils (fibres)
•Clump together to
make a plaque
•Prion protein (PrP)
 Prion Fibrils

View of purified protein under an electron

microscope
 Neurodegenerative diseases with
similarities
• Prion diseases (TSEs) – prion protein
• Alzheimer’s disease - amyloid-β
• Parkinson’s disease - α-synuclein
• Huntington’s disease – huntingtin
• Wilson’s disease – defects in copper metabolism
• Amyotrophic lateral sclerosis – defects in
superoxide dismutase
But prion diseases can be
transmissible…Kuru
 But prion diseases can be
transmissible…

BSE (mad cow disease) to variant Creutzfeldt-Jakob disease

(vCJD)
vCJD
• BSE noted in UK cattle 1986
• Government denied a risk to public till
early 1990s
• 3 vCJD deaths in 1995, last in 2016
• Deaths 123 definite, 55 probable in
UK John Gummer, Minister
for Agriculture, Fisheries
• Median age of onset 26.5 years, age and Food (89-93) gives
of death 28 years his daughter a ‘safe’
• 42% female / 58% male beef burger in May 1990
 Transmission - evidence
• Injections of diseased brain tissue into another
animal of the same species transmits the disease
• Suggests an infectious agent such as a virus
• But, no evidence of a virus found in brain extracts
• Treating the extracts with agents (e.g. ultraviolet
light/nucleases) that destroy nucleic acids does not
reduce their infectiousness
• The evidence indicates that the infectious agent in
the TSEs is a protein
An infectious protein?

• Prion: proteinaceous infectious particle

• PrPSc - scrapie form of the prion protein
• (sometimes called PrPRes but ‘resistant’ form may
not always be infectious)
• Protein in the amyloid plaques
• Prion diseases are more likely to be sporadic or
inherited
Routes of infection
• Ingestion (kuru, vCJD)
• Iatrogenic
• Corneal grafts
• Dura mater grafts
• Human derived
growth hormone
injection (hGH)
• Experimental
transmission in animal
models

Aguzzi A., Nuvolone M. & Zhu C. (2013)

Nature Reviews Immunology, 13, pages 888–902
From mouth to brain
• Prions detected by
immune system
• Resistant to proteolysis
• Replicate in lymphoid
tissue (especially
follicular dendritic cells)
• Enter central nervous
system by nerves of the
autonomic nervous
system

Aguzzi A., Nuvolone M. & Zhu C. (2013)

Nature Reviews Immunology, 13, pages 888–902
 Prions, Health and Disease
• There is a normal cellular form of the prion protein
PrPC
• PrPC & PrPSc share the same amino acid
sequence
• Coded by prnp gene on xsome 20 (human)
• PrPC to PrPSc conversion is a key event in disease
• prnp -/- (gene knock out) cannot be infected
• i.e. PrP expression is essential for disease
pathogenesis
• Knockout phenotype shows disturbed sleep
patterns and sensitivity to oxidative stress
 The Essence of Prion Disease

Conversion of normal cellular prion protein to

an abnormal form
????
PrPC PrPSc
PrPSc
GPI anchored (plasma Extracellular
membrane) Aggregated
Monomeric Protease resistant
Protease sensitive No specific copper bound
Binds copper High beta sheet content
Alpha helices present Associated with infectivity
Synaptic location Not easily decontaminated
Conformations of PrP

PrPC PrPSc
3 α–helices Secondary structure
2 β-strands β-strand rich
NMR & X-ray Model shown
Proposed mechanisms of PrPSc
catalysed conversion
Template directed refolding model

Nucleation dependent polymerisation model

PrPSc in Scrapie

30 60 90 110 120 130 166

Days Post Infection

Mechanism of neurodegeneration

• PrPSc is cytotoxic
• Potential loss of antioxidant activity of PrPC
• Apoptotic neurones cleared by microglia
• Leave ‘gaps’ (sponge like) and protein plaques
Aguzzi A., Nuvolone M. & Zhu C. (2013)
Nature Reviews Immunology, 13, pages 888–902
Conformations of PrP

PrPC PrPSc
PrPSc
But what causes initial formation of PrPSc ?
Inherited prion diseases

Mutations destabilise PrPC / make PrPSc more

energetically favourable
 Sporadic Prion Diseases
• No family history of the disease / No known exposure to
infectious prions and late onset
• Cause is uncertain, possibly:
• a spontaneous somatic mutation may have occurred in
one of the prnp genes in a cell.
• normal PrPC protein may have spontaneously converted
into the PrPSc form.
• Tends to be a susceptibility polymorphism in their prnp genes:
• vCJD: all but one case was Met/Met homozygous at codon 129
• Selective breeding of sheep to remove susceptible alleles
 Guidance when treating patients
• Detailed guidance is published by the government
• Precautions for patients with, or “at increased risk” of, CJD, undergoing
surgical procedures includes:
• perform the intervention in an operating theatre
• schedule at the end of the list, to allow cleaning
• involve the minimum number of personnel required
• Protective clothing should be worn, i.e. liquid repellent operating
gown, over a plastic apron, gloves, mask and goggles, or full-face
visor; for symptomatic patients, this protective clothing should be
single-use
• Single-use disposable surgical instruments and equipment should be
used where possible, and subsequently destroyed by incineration or
sent to the instrument store;
• Effective tracking of reusable instruments should be in place, so that
instruments can be related to use on a particular patient.
•https://www.gov.uk/government/publications/guidance-from-the-
acdp-tse-risk-management-subgroup-formerly-tse-working-group
 Blood & blood products
• Safety measures to reduce any possible risk of
spreading vCJD through blood:
• withdrawal and recall of any blood components donated
by anyone who develops vCJD & importing plasma from
the USA
• removing white blood cells from all blood used for
transfusions (leucodepletion)
• not accepting donations from people who may have
received a blood transfusion since 1980
• promoting the appropriate use of blood, tissues and
alternatives throughout the NHS - led to a reduced
amount of blood transfused during and following surgery
•https://assets.publishing.service.gov.uk/government/uploads/system/uplo
ads/attachment_data/file/727300/Background_information_for_healthcar
e_staff.pdf
Potential pharmaceutical interventions

From Aguzzi A.,

Nuvolone M. & Zhu C.
(2013)
Nature Reviews
Immunology, 13, pages
888–902
 Prion diseases
• Fatal neurodegenerative diseases
• An unexpected infectious agent - Protein
• Conformation change:
• normal cellular → pathogenic form
• Long incubation time – asymptomatic
• Resistant to normal sterilisation and
decontamination procedures