EUROPE GOES ALTERNATIVE

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• EUROPE GOES ALTERNATIVE
• Animal Testing and Alternative Methods -
• a Scientific Perspective
• Coenraad F.M.Hendriksen
• Netherlands Centre Alternatives to Animal Use
  (NCA), Utrecht University
• Netherlands Vaccine Institute (NVI)

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Animal Testing and Alternative Methods - a
scientific perspective

• Topics of presentation
• science and alternatives -
• the way to go!
• Other reasons for going
• alternative
• 3Rs recommendations
• Conclusion

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
3
Science and Alternatives - the way to go!
NCA The Netherlands Centre Alternatives to
Animal Use
(Timo Breit)
Scientific and regulatory evolution from
what to why (1)
Until now questions are based on what -
what is the toxicity of a chemical? - what
is the potency of a vaccine? - what is the
safety of a pharmaceutical?
Future why questions will dominate -
why is a chemical toxic? - why is a batch
of vaccine potent? - why is a
pharmaceutical not safe?
Conference Europe Goes Alternative, Brussels,
071105
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Science and Alternatives - the way to go!
Paradigm shift in Science and regulatory
testing from what to why (2)
What questions Parameter Endpoint (animal
model) vaccine potency protection after
challenge chemical toxicity increase tumour
frequency drug safety clinical signs
Why questions Endpoint Model (e.g.) 3-D
configuration antigen physiochemical DNA or DNA
repair change at molecular level tissue
culture interaction at receptor level tissue
culture
NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
5
New developments
Science and Alternatives - the way to go!
NCA The Netherlands Centre Alternatives to
Animal Use

• Examples
• - omics technologies bioinformatics
• embryonic stem cells
• in silico modelling tools
• improvements in tissue culture techniques
• systems biology
• etc.

Conference Europe Goes Alternative, Brussels,
071105
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Science and Alternatives - the way to go!
Paradigm shift in science and regulatory testing
from what to why (3)
Past/present
Future
Conclusion Future will give a shift in research
and regulatory testing paradigms non-animal
models will increasingly being used to generate
answers to the why questions (mechanistic info)
while the animal model will be used to verify and
confirm data from non-animal studies
R D start
NCA The Netherlands Centre Alternatives to
Animal Use
End of research objective
Conference Europe Goes Alternative, Brussels,
071105
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Other reasons for Europe to go alternative

• Animal use concerns
• Moral concerns
• Economic and pragmatic concerns
• Scientific concerns
• (In)validation concerns

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
8
Other reasons for Europe to go alternative
Societies acceptance of animal use for specific
purposes

• Animal use concerns
• Moral concerns
• Economic and pragmatic concerns
• Scientific concerns
• (In)validation concerns

NCA The Netherlands Centre Alternatives to
Animal Use
(Aldhous et al. 1999)
Conference Europe Goes Alternative, Brussels,
071105
9
Other reasons for Europe to go alternative

• Animal use concerns
• Moral concerns
• Economic and pragmatic concerns
• Scientific concerns
• (In)validation concerns

• Increased legislation and regulation concerning
  laboratory animal use
• Revision of Council
• Directive 86/609/EEC
• Appendix A of the
• European Convention

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
10
Other reasons for Europe to go alternative

• Animal use concerns
• Moral concerns
• Economic and pragmatic concerns
• Scientific concerns
• (In)validation concerns

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
11
Europe goes alternative - Future trend in Lab.
Animal use?
NCA The Netherlands Centre Alternatives to
Animal Use
year
year
Conference Europe Goes Alternative, Brussels,
071105
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Europe goes Alternative the way forward

• Objective Conference
• demonstrate that the European Union keeps animal
  welfare high on the political agenda and remains
  in the lead for animal protection,
• show industrys approach to actively promote
  research in alternative methods,
• illustrate progress made in the area of
  alternative methods to animal tests at EU and
  intermational level,
• identify further possibilities to improve
  development, validation and legal acceptance of
  alternative methods

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
13
Objective identify further possibilities to
improve development, validation and legal
acceptance of alternative methods
Effective EU policy towards Europe goes
Alternative should include the following
elements
Rex Burch
Bill Russell
R
eality
NCA The Netherlands Centre Alternatives to
Animal Use
R
ealism
R
eaction
Conference Europe Goes Alternative, Brussels,
071105
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Reality numbers and purposes of animal use
Focus of Commission concerning 3Rs research
priorities
NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
15
Reality numbers and purposes of animal use
No. of animals used in EU-MS in 2002 (N total
10.731.020) purposes
NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
16
Reality alternatives and the 3Rs principle
Replacement Desirable as replacement is, it
would be a mistake to put all our humanitarian
eggs in this basket alone. (Russell Burch,
1959)
Replacement of animal tests by methods not using
animals
Reduction reducing the numbers of animals needed
to answer a scientific question
Reduction .of all modes of progress it is the
one most obviously, immediately, and universally
advantageous in terms of efficiency. (Russell
Burch, 1959)
NCA The Netherlands Centre Alternatives to
Animal Use
Refinement reducing pain and distress in a
particular animal experiment
Refinement Happy animals make good
science. (Trevar Poole)
Conference Europe Goes Alternative, Brussels,
071105
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Objective identify further possibilities to
improve development, validation and legal
acceptance of alternative methods
Reality
Realism
NCA The Netherlands Centre Alternatives to
Animal Use
Reaction
Conference Europe Goes Alternative, Brussels,
071105
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Realism Proposed timeframe for full replacement
of animal tests

• Human Health effects Foreseeable time
  involved
• for full replacement
• _________________ _____________________
• acute toxicity gt 2014
• skin irritation gt 2014
• skin corrosion lt 2004
• eye irritation gt 2010
• skin sensitisation gt 2019
• skin absorption/penetration gt 2006
• subacute/subchronic toxicity gtgt 2014 (not
  foreseeable)
• genotoxicity/mutagenicity gt 2016
• UV-induced effects gtgt 2019 (not foreseeable)
• Toxicokinetics and biotransformation gtgt 2014
  (not foreseeable)
• Carcinogenicity gtgt 2014 (not foreseeable)
• Reproductive toxicity gtgt 2014 (not
  foreseeable)

NCA The Netherlands Centre Alternatives to
Animal Use
(SCCNFP, 2004)
Conference Europe Goes Alternative, Brussels,
071105
19
Realism Legislative and Regulatory context

• Sectorial regulations and decisions regulate
• the activities of particular sector.
• Horizontal legislation interacts with
• aspects of sectorial regulations

• Examples of conflict between sectorial European
  directives and EU legislation 86/609/EEC
• re-testing of biological products entering the
  European market
• shellfish toxin testing policies
• Other concerns
• Slow progress in Harmonisation and/or mutual
  acceptance
• Increased priority for risk minimisation

Regulations to animal testing and multilateral
agreements, e.g. Council Directive 86/609/EEC
NCA The Netherlands Centre Alternatives to
Animal Use
Horizontal legislation
Council Directive pharmaceuticals
Council Directive vaccines
Council Directive pesticides
Council Directive chemicals
Sectorial regulation
(According to Schiffelers et.al.2005)
Conference Europe Goes Alternative, Brussels,
071105
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Objective identify further possibilities to
improve development, validation and legal
acceptance of alternative methods
Reality
Realism
NCA The Netherlands Centre Alternatives to
Animal Use
Reaction
Conference Europe Goes Alternative, Brussels,
071105
21
Reaction obstacles concerning implementation of
validated alternatives

• Council Directive 86/609/EEC on the protection of
  laboratory animals
• Art. 7 (2)
• An experiment shall not be performed if another
  scientifically satisfactory method of obtaining
  the results sought, not entailing the use of an
  animal, is reasonably and practically available
• Art. 7 (3)
• In a choice between experiments, those which use
  the minimum number of animals, cause the least
  pain, suffering, distress or lasting harm and
  which are most likely to provide satisfactory
  results shall be selected.

The difficulty lies not in the new ideas, but in
escaping from the old ones (John Maynard Keynes,
1883-1946)
NCA The Netherlands Centre Alternatives to
Animal Use
Examples - European Pharmacopoeia monographs
for Diphtheria and Tetanus vaccine specify two
possibilities for potency testing, substantially
differing in the level of suffering from each
other - Production of monoclonal antibodies
Conference Europe Goes Alternative, Brussels,
071105
22
Europe goes Alternative the way forward

• Objective Conference
• demonstrate that the European Union keeps animal
  welfare high on the political agenda and remains
  in the lead for animal protection,
• show industrys approach to actively promote
  research in alternative methods,
• illustrate progress made in the area of
  alternative methods to animal tests at EU and
  international level,
• identify further possibilities to improve
  development, validation and legal acceptance of
  alternative methods

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
23
Alternatives developed and validated for
regulatory testing in the last decade

• Chemicals/biologicals
• Animal model Alternative model
• skin corrosivity test tissue
  culture tests
• phototoxicity tests cell culture
  test
• embryotoxicity tests cell culture
  test
• skin sensitization test LLNA test
• potency testing of tetanus Elisa test and
  ToBI test
• potency test erysipelas Elisa test
• vaccine
• target animal safety test deletion
• monoclonal antibody in vitro culture
• production

Chemicals/biologicals Animal model
Alternative model
NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
24
Alternatives developed and validated for
regulatory purposes in the last decade

• Chemicals/biologicals
• Animal model Alternative model
• skin corrosivity test tissue
  culture tests
• phototoxicity tests cell culture
  test
• embryotoxicity tests cell culture
  test
• skin sensitization test LLNA test
• potency testing of Elisa test, ToBI test
• tetanus vaccine
• monoclonal antibody in vitro production
• production
• classical LD50 tests 3 alternative
  approaches
• pyrogenicity testing LAL test and in vitro
  model
• percutaneous adsorption cell culture test
• production inactivated in vitro
  culture
• polio vaccine (monkeys)
• abnormal toxicity test test deleted
• specific tox, diphtheria cell culture
  test

• Chemicals/biologicals
• Animal model Alternative model
• potency test hepatitis antigen
  quantification
• vaccine
• neurovirulence test PCR transgenic animals
• polio vaccine
• potency test erysipelas Elisa test
• vaccine
• potency test various clos- Elisa tests
• tridial vaccines
• safety test erysipelas test deleted
• vaccine
• potency test leptospirosis antigen
  quantification
• vaccine
• target animal safety test
• safety testing poultry in vitro
  methods
• vaccine

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
25
Alternatives developed and validated for
regulatory purposes in the last decade (tests
developed in Europe are given in blue)

• Chemicals/biologicals
• Animal model Alternative model
• skin corrosivity test tissue
  culture tests
• phototoxicity tests cell culture
  test
• embryotoxicity tests cell culture
  test
• skin sensitization test LLNA test
• potency testing of Elisa test, ToBI test
• tetanus vaccine
• monoclonal antibody in vitro production
• production
• classical LD50 tests 3 alternative
  approaches
• pyrogenicity testing LAL test and in vitro
  model
• percutaneous adsorption cell culture test
• production inactivated in vitro
  culture
• polio vaccine (monkeys)
• abnormal toxicity test test deleted
• specific tox, diphtheria cell culture
  test

• Chemicals/biologicals
• Animal model Alternative model
• potency test hepatitis antigen
  quantification
• vaccine
• neurovirulence test PCR transgenic animals
• polio vaccine
• potency test erysipelas Elisa test
• vaccine
• potency test various clos- Elisa tests
• tridial vaccines
• safety test erysipelas test deleted
• vaccine
• potency test leptospirosis antigen
  quantification
• vaccine
• target animal safety test
• safety testing poultry in vitro
  methods
• vaccine

NCA The Netherlands Centre Alternatives to
Animal Use
Conference Europe Goes Alternative, Brussels,
071105
26
Europe goes alternative!
Vision without action is daydream Action
without vision is nightmare --Japanese proverb

• Recommendation an independent advisory board to
  the Commission to identify
• critical aspects of animal use,
• priorities in 3Rs research
• potential obstacles concerning implementation,
  and
• potential conflicts between 86/609/EEC and
  other Directives

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Thanks for your attention!!
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(No Transcript)
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if so, why do we still use animals?
Validated non-animal tests Only 9 non animal
tests endorsed as scientifically valid by
ECVAM skin corrosivity (Rat TER, CORROSITEX,
Episkin, Epiderm), phototoxicity (3T3NRU),
embryotoxicity (EST, MM, whole embryo assay),
in vitro percutaneous absorption.
NCA The Netherlands Centre Alternatives to
Animal Use
Annual European Meeting Toxicology Forum,
Brussels, 091105
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Ontwikkelingen in het gebruik van proefdieren
(De Boo)
NCA jubileum symposium, 25 november 2004