| Toxicity Endpoint |
Number
of Methods |
Test Method [number] |
Regulatory Application and ICCVAM Recommendations |
|
Acute Systemic Toxicity |
5 |
Up-and-Down Procedure (UDP) |
In 2001, ICCVAM recommended the UDP as a replacement alternative for OECD Test Guideline (TG) 401,
the traditional in vivo rodent LD50 test for assessing acute oral systemic toxicity.
The UDP was adopted by OECD as TG 425 in 2003. |
|
In vitro basal cytotoxicity methods [2] |
In 2007, ICCVAM recommended both in vitro test methods as reduction alternatives to estimate
the starting dose in the UDP and Fixed Dose Procedure for assessing acute oral systemic
toxicity. Recommendations were accepted by U.S. Federal agencies. |
| 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 HepaRG cells and cryopreserved human hepatocytes. |
| Biologics Testing |
24 |
- 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
NICEATM-ICCVAM/ECVAM-sponsored
workshop and future activities recommended. |
| In vitro potency test for Leptospirosis (1) |
One ICCVAM Agency, USDA, has an ongoing validation study in conjunction with the University of
Michigan on an in vitro potency test for a Leptospirosis vaccine. |
| Developmental Toxicity |
1 |
Frog Embryo Teratogenesis Assay: Xenopus (FETAX) |
In 2000, FETAX was reviewed at a NICEATM-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. |
| 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, to be included in EPA’s
Endocrine Disruptor Screening Program. In 2003, a report with guidance
for protocol standardization and validation studies released; in 2006, reference substance list was revised. |
- In vitro estrogen receptor (ER) binding [14]
- In vitro ER TA [95]
|
Same as for in vitro AR assays. NICEATM-ICCVAM are sponsoring an ongoing
international validation study of an in vitro ER TA assay, and is working with the test method
developer to develop and implement validation study protocols for a second in vitro ER TA
assay. |
| Eye Corrosion/Irritation |
17 |
In vitro test methods
for detecting ocular corrosives and severe irritants [4] |
In 2007, the Bovine Corneal Opacity and Permeability (BCOP) and the Isolated Chicken Eye test methods
were recommended as screening tests for identifying corrosives and severe irritants, with certain
limitations. Two other methods were not recommended for regulatory hazard classification purposes until further
developed and evaluated. |
| In vitro test methods for assessment
of the eye irritation potential of antimicrobial cleaning products [3] |
An approach using the BCOP, the EpiOcular and the Cytosensor Microphysiometer test methods for
evaluating the eye irritation potential of certain antimicrobial cleaning products is
currently under review. |
| In vitro tissue-based test
methods for detecting mild to moderate irritants and nonirritants [4] |
The four tissue-based in vitro methods evaluated by ICCVAM for detection of ocular
corrosives (described above) are currently being evaluated by ICCVAM for their utility for
identification of mild to moderate irritants and substances not labeled as ocular
irritants. |
| 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, cytosensor microphysiometer and red blood cell haemolysis test methods) for
identification of mild to moderate irritants and substances not labeled as ocular irritants are
currently being reviewed by ICCVAM for U.S. regulatory applicability. |
| 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. |
| Genetic Toxicity |
4 |
In vitro mammalian cell micronucleus test |
NICEATM and the ICCVAM Genetic
Toxicity Working Group (GTWG) are involved in development of a draft OECD Test Guideline
and have provided comments on a study to determine the most appropriate measure of cytotoxicity
for inclusion in the Test Guideline. |
| In vivo rodent alkaline
comet assay for detection of genotoxic carcinogens |
NICEATM and the GTWG are 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 |
The four tissue-based in vitro methods evaluated by ICCVAM for detection of ocular
corrosives (described above) are currently being evaluated by ICCVAM for their utility for
identification of mild to moderate irritants and substances not labeled as ocular
irritants. |
| 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; also provided nominations of independent experts to serve on an ESAC peer
review panel. |
| Pyrogenicity |
5 |
In vitro pyrogenicity |
In October 2008, ICCVAM recommended five in vitro pyrogenicity test methods
measuring cytokine release from human cells as replacements
for the rabbit test, subject to product-specific validation, to detect endotoxin
contamination in parenteral drugs. Responses from agencies due April 2009. |
| Skin Corrosion |
4 |
- 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 DOT;
otherwise, recommended as part of a tiered testing strategy; in 2000, accepted by U.S. agencies; in 2006, adopted by OECD
as TG 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 based on three in vitro tests. An expert
consultation hosted by U.S. is scheduled for June 15-17, 2009. |
| Skin Sensitization |
7 |
Murine Local Lymph Node Assay (LLNA)
- Reduced LLNA (rLLNA)
- Use for potency determination
- Applicability domain
- Performance standards
|
In 1999, the LLNA was recommended by ICCVAM and accepted by
regulatory agencies as alternative for guinea pig tests for allergic contact dermatitis;
adopted in 2002 as TG 429 by OECD. In 2009, ICCVAM recommended the rLLNA to regulatory
agencies and finalized performance standards for the LLNA. Recommendations included an
updated protocol that uses fewer animals. ICCVAM has determined that the LLNA may be useful
in determining the relative potency of sensitizers as part of a weight-of-evidence approach.
ICCVAM recommendations for the LLNA applicability domain and three non-radiolabeled LLNA
methods are currently being finalized. |
| LLNA non-radiolabeled methods [3] |
In vitro approaches
- In vitro cell-based methods [2]
- Peptide reactivity assay
|
The human cell line activation test (h-CLAT), the myeloid U937 skin sensitization test
(MUSST), and the dipeptide reactivity assay are under consideration for further validation
studies. ICCVAM is participating in the Validation Management Group with ECVAM and JaCVAM. |
| Total |
212 |
