| Toxicity Endpoint |
Number
of Methods |
Test Method [number] |
Regulatory Application and ICCVAM Recommendations |
|
Acute Systemic Toxicity |
6 |
Up-and-down procedure (UDP)
for acute oral toxicity |
In 2001, ICCVAM recommended the revised UDP as a replacement alternative for
the traditional in vivo rodent LD50 test for assessing acute oral systemic toxicity.
The updated UDP was adopted by OECD as TG 425 in 2003. |
|
In vitro basal cytotoxicity methods [2] |
In 2007, ICCVAM recommended two in vitro test methods as reduction alternatives to estimate
the starting dose in the UDP, the acute toxic class method, and the fixed dose procedure for assessing acute oral systemic
toxicity. Recommendations were accepted by U.S. Federal agencies. OECD Guidance Document 129 for
implementation of the test methods was published in 2010. |
| Biotransformation enzyme induction assays [2] |
NICEATM and ICCVAM participants are
providing input and guidance to an ECVAM Validation Study of a human hepatic biotransformation
enzyme induction assay using cryopreserved HepaRG cells and human hepatocytes. This project is ongoing. |
| UDP for acute dermal toxicity |
NICEATM and the ICCVAM Interagency Acute Toxicity Working Group are
developing the dermal UDP as a potential replacement alternative
for OECD TG 402, the traditional in vivo rodent LD50 test for assessing acute
dermal systemic toxicity. This project is ongoing. |
| Biologics Testing |
28 |
- In vivo alternatives
- Ex vivo alternatives
- In vitro cell-based methods
- In vitro enzymatic alternatives
|
In 2006, various reduction, refinement, and replacement alternatives to the
mouse LD50 assay for botulinum toxin detection and potency testing were reviewed at a
ICCVAM
workshop and future activities recommended. |
| In vitro ELISA replacement potency release tests for veterinary Leptospira
vaccines [4] |
In vitro ELISA antigen quantification methods for potency determination of four veterinary
Leptospira vaccines were reviewed at a 2010 ICCVAM workshop and future activities
recommended. These tests replace the vaccination-challenge test previously performed in
hamsters. |
| Serology potency test for veterinary rabies vaccines |
The serum neutralization test approved by the European Pharmacopoeia Commission to
replace the vaccination-challenge test in mice was reviewed at a 2011 ICCVAM
workshop and future activities recommended. |
| Developmental Toxicity |
1 |
Frog embryo teratogenesis assay: Xenopus (FETAX) |
FETAX was reviewed at a 200 ICCVAM sponsored workshop as a reduction or replacement
alternative to assess the developmental toxicity of chemicals and mixtures. Data gaps and
inadequacies were identified and future activities recommended. |
| Eye Corrosion/Irritation |
20 |
In vitro test methods
for detecting ocular corrosives and severe irritants [4] |
ICCVAM recommended the bovine corneal opacity and permeability (BCOP) and the isolated chicken eye (ICE) test methods
as screening tests for identifying corrosives and severe irritants, with certain
limitations. Recommendations were accepted by U.S. Federal agencies in 2008.
Two other methods were not recommended for regulatory hazard classification purposes until further
developed and evaluated. OECD Test Guidelines for BCOP (TG 437) and ICE (TG 438) are now
available. |
| Use of topical anesthetics, systemic analgesics, and humane endpoints
in in vivo ocular safety testing [1] |
ICCVAM recommendations on routine use of topical anesthetics, systemic analgesics, and humane endpoints
to avoid or minimize pain and distress during required in vivo ocular safety testing were accepted
by Federal agencies in 2011. |
| In vitro test methods for assessment
of the eye irritation potential of antimicrobial cleaning products [3] |
ICCVAM evaluated an approach using the BCOP, the EpiOcular and the Cytosensor microphysiometer (CM) test methods
to assess the eye irritation potential of certain antimicrobial cleaning products.
ICCVAM recommendations for future studies were accepted
by Federal agencies in 2011. |
| In vitro tissue-based test
methods for detecting mild to moderate irritants and nonirritants [4] |
ICCVAM recommended that these four in vitro test methods must be improved
before they can be used in regulatory safety testing to classify substances as having
the potential to cause reversible, nonsevere eye injuries or as not requiring hazard
labeling for eye irritation. The ICCVAM recommendations were accepted by U.S. Federal agencies in March 2011. |
| In vitro cell function-based test
methods for detecting mild to moderate irritants and nonirritants [4] |
ECVAM evaluations of four cell function-based in vitro methods (fluorescein leakage,
neutral red release, CM and red blood cell haemolysis test methods) for
classification of ocular hazards have been reviewed
by ICCVAM for U.S. regulatory applicability.
ICCVAM recommendations on use of the CM
test method for classification of ocular hazards
were accepted by U.S. Federal agencies in March 2011. |
| Low volume eye test (LVET) [1] |
ICCVAM recommended to Federal agencies that the LVET should not be used for future regulatory
testing due to poor predictivity when compared to the current standard rabbit eye test. However,
data from past LVET studies may be considered in a weight-of-evidence
approach to classify ocular hazards. The ICCVAM recommendations were accepted by U.S. Federal agencies in March 2011. |
| Recombinant human tissue models [2] |
NICEATM and ICCVAM representatives are serving on the Validation Management Group for a
prospective validation of reconstructed human tissue models (EpiOcular and
SkinEthic) for identification of mild to moderate irritants and substances not labeled as ocular irritants. |
| Short time exposure (STE) test method |
As part of the ICATM collaboration, JaCVAM requested that ICCVAM conduct an international
peer review of the STE test method, which assesses eye irritation potential by measuring
cytotoxicity in rabbit corneal epithelial cells. NICEATM will prepare a summary review
document on the validation status of the STE to be considered by the ICCVAM peer review
based on a BRD provided by the test method developer. |
| Endocrine Disruptors |
138 |
- In vitro androgen receptor (AR) binding [11]
- In vitro AR transcriptional activation (TA) [18]
|
In 2002, ICCVAM evaluated screens for identifying
potential endocrine-disrupting chemicals. A 2003 report
based on that evaluation provided guidance
for protocol standardization and validation studies.
A 2006 addendum to the report provided a revised reference substance list. |
- In vitro estrogen receptor (ER) binding [14]
- In vitro ER TA [95]
|
These assays were also addressed in the 2003 report and 2006 addendum. ICCVAM coordinated validation studies of
two in vitro test methods used to detect estrogenic and anti-estrogenic activities: the
BG1Luc ER TA test method (also known as the LUMI-CELL ER assay)
developed by XDS, Inc., and the MCF-7 cell
proliferation assay developed by CertiChem, Inc. In 2012, Federal agencies accepted
ICCVAM recommendations that the BG1Luc ER TA
test method could be used as a screening test to identify substances with in vitro estrogen receptor agonist
and/or antagonist activity. Data from the validation study of the CertiChem MCF-7 assay is currently being evaluated. |
| Genetic Toxicity |
4 |
In vitro mammalian cell micronucleus test |
NICEATM and the ICCVAM Interagency Genetic
Toxicity Working Group (GTWG) were involved in development of OECD Test Guideline 487:
In Vitro Mammalian Cell Micronucleus Test, published in 2010. |
| In vivo rodent alkaline
comet assay for detection of genotoxic carcinogens |
NICEATM and the GTWG were involved in development of the validation study plan, the proposed
protocol, and proposed list of reference substances, and have representatives on the Validation
Study Management Team. |
| In vitro TK6
alkaline comet assay |
NICEATM and the GTWG were involved in development of the validation study plan, the proposed
protocol, and proposed list of reference substances, and have representatives on the Validation
Study Management Team. |
| Cell transformation assay |
NICEATM and the
GTWG provided comments to JaCVAM on their validation study plan and protocol for their
validation study, as well as providing liaison members to the Validation Study
Management Team; provided nominations of independent experts to serve on an ESAC peer
review panel;
also provided comments to ECVAM and U.S. National Coordinator on proposed OECD Test Guideline. |
| Pyrogenicity |
5 |
In vitro pyrogenicity test methods (monocyte activation test [MAT] and four other test methods) |
In 2008, ICCVAM recommended five in vitro pyrogenicity test methods
measuring cytokine release from human cells as replacements
for the rabbit pyrogen test to detect endotoxin contamination in parenteral drugs,
subject to product-specific validation. All applicable Federal agencies accepted or
endorsed the ICCVAM recommendations in May 2009. BiotestAG is preparing a comprehensive
BRD for NICEATM to consider the validation status of the MAT for identifying nonendotoxin
pyrogens. The MAT was one of the five test methods accepted by Federal agencies in 2009;
the BiotestAG BRD will include results of studies performed in response to ICCVAM recommendations. |
| Skin Corrosion |
5 |
- Corrositex®
- EpiDerm™
- EPISKIN™
-
Rat Transcutaneous Electrical Resistance (TER) Assay
- SkinEthic Assay
|
In 1999, ICCVAM recommended Corrositex® as a stand-alone assay for evaluating acids, bases,
and acid derivatives for the U.S. Department of Transportation;
otherwise, recommended as part of a tiered testing strategy; in 2000, accepted by U.S. agencies; in 2006,
adopted by OECD
as Test Guideline 435. In 2002, TER and human skin models recommended as part of a tiered testing strategy; in
2004, adopted by OECD as TG 430/431. |
| Skin Irritation |
3 |
- EpiDerm™
- EPISKIN™
- SkinEthic Assay
|
In 2008, OECD Test Guidelines were proposed for three in vitro tests. An expert
consultation hosted by U.S. took place in 2009, and the test methods were adopted by OECD as TG 439 in 2010. |
| Skin Sensitization |
10 |
Murine local lymph node assay (LLNA)
- Reduced LLNA (rLLNA)
- Performance standards
- Applicability domain
- Use for potency categorization
|
- In 1999, the LLNA was recommended by ICCVAM
and accepted by
regulatory agencies as an alternative for guinea pig tests for allergic contact dermatitis
hazard testing. The LLNA was
adopted in 2002 as Test Guideline 429 by OECD.
- In 2009, ICCVAM made recommendations to Federal agencies on performance standards for the LLNA,
an updated protocol that uses fewer animals, and use of the rLLNA to regulatory agencies.
Federal agencies accepted the ICCVAM recommendations in March 2010. ICCVAM recommendations were incorporated
into an updated OECD Test Guideline 429 published in July 2010.
- In 2010, ICCVAM made recommendations to Federal agnencies
for the LLNA applicability domain and two nonradioactive LLNA
methods. Federal agencies accepted the ICCVAM recommendations in February 2011.
OECD test guidelines for the two
nonradioactive methods (Test Guidelines 442A and 442B) incorporate ICCVAM
recommendations. ICCVAM deferred a formal recommendation on a third
nonradioactive method pending the receipt of additional data.
- In 2011, ICCVAM recommended to Federal agencies that the LLNA may be used to
categorize substances as strong sensitizers. Federal agencies accepted the ICCVAM recommendations in
February 2012.
|
| LLNA nonradioactive methods [3] |
In vitro approaches
- In vitro cell-based methods [4]
- Peptide reactivity assay
|
The KeratinoSens assay and the dipeptide reactivity assay (DPRA) are undergoing peer review at ECVAM.
Validation activities are underway for the human cell line activation test (h-CLAT), the myeloid U937
skin sensitization test (MUSST), and the IL-8 Luc assay. ICCVAM is participating on the Validation
Management Teams with ECVAM and JaCVAM. |
| Total |
220 |
