[Federal Register Volume 76, Number 9 (Thursday, January 13, 2011)]
[Notices]
[Pages 2388-2390]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 2011-669]


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DEPARTMENT OF HEALTH AND HUMAN SERVICES


National Toxicology Program (NTP); NTP Interagency Center for the 
Evaluation of Alternative Toxicological Methods (NICEATM); Federal 
Agency Responses to Interagency Coordinating Committee on the 
Validation of Alternative Methods (ICCVAM) Recommendations on Two 
Nonradioactive Versions of the Murine Local Lymph Node Assay (LLNA) for 
Assessing Allergic Contact Dermatitis (ACD) Hazard Potential of 
Chemicals and Products, and Expanded Uses of the LLNA for Pesticide 
Formulations and Other Products; Notice of Availability

AGENCY: National Institute of Environmental Health Sciences (NIEHS), 
National Institutes of Health (NIH), HHS.

ACTION: Notice of Availability.

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SUMMARY: U.S. Federal agency responses to ICCVAM test method 
recommendations on two nonradioactive versions of the LLNA for 
assessing the ACD hazard potential of chemicals and products and for 
expanded uses of the LLNA for pesticide formulations and other products 
are now available on the NICEATM-ICCVAM Web site at http://iccvam.niehs.nih.gov/methods/immunotox/llna.htm. ICCVAM recommended the 
nonradioactive LLNA: 5-bromo-2-deoxyuridine-enzyme-linked immunosorbent 
assay

[[Page 2389]]

(BrdU-ELISA) and LLNA: Daicel Adenosine Triphosphate (DA), and expanded 
uses for the LLNA. In accordance with the ICCVAM Authorization Act (42 
U.S.C. 285l-3(e)(4)), ICCVAM forwarded recommendations to Federal 
agencies and made these recommendations available to the public (75 FR 
37443). Agencies have now notified ICCVAM in writing of their findings 
and ICCVAM is making these responses available to the public.

FOR FURTHER INFORMATION CONTACT: Dr. William S. Stokes, Director, 
NICEATM, NIEHS, P.O. Box 12233, Mail Stop: K2-16, Research Triangle 
Park, NC 27709, (telephone) 919-541-2384, (fax) 919-541-0947, (e-mail) 
[email protected]. Courier address: NICEATM, NIEHS, Room 2034, 530 
Davis Drive, Morrisville, NC 27560.

SUPPLEMENTARY INFORMATION:

Background

    In 1999, ICCVAM recommended the LLNA as a valid safety test for 
assessing the ACD hazard potential of many chemicals and products (NIH 
Publication No. 99-4494; available at http://iccvam.niehs.nih.gov/methods/immunotox/llna_PeerPanel98.htm). ICCVAM also concluded that 
the LLNA, when used as an alternative method to the guinea pig 
maximization test (GPMT) or the Buehler test (BT), could also 
significantly reduce animal use and improve animal welfare. Based on 
this evaluation, the U.S. Environmental Protection Agency (EPA 2003), 
the U.S. Food and Drug Administration, and the U.S. Consumer Product 
Safety Commission (CPSC) subsequently accepted the method as a valid 
substitute for the GPMT and BT (http://iccvam.niehs.nih.gov/methods/immunotox/llna_PeerPanel98.htm). The Organisation for Economic Co-
operation and Development (OECD) subsequently adopted the LLNA in 2002 
as international OECD Test Guideline 429 (OECD, 2002). The 
International Organization for Standardization (ISO) adopted the LLNA 
as ISO standard 10993-10 in 2002 (ISO, 2002).
    ICCVAM recommended an updated LLNA test method protocol in 2009 
that further reduced animal use for each safety test by 20-40% (ICCVAM, 
2009). Federal agencies endorsed this updated protocol (75 FR 25866). 
OECD Test Guideline 429 was subsequently updated in 2010 to incorporate 
the updated revisions (OECD, 2010a). The ISO standard was also updated 
in 2010 (ISO, 2010).
    Compared to the LLNA, the LLNA: BrdU-ELISA and LLNA: DA do not use 
radioactive reagents and therefore provide additional advantages in 
terms of reduced hazardous waste disposal and broader availability for 
use by laboratories that cannot use radioactive reagents. ICCVAM 
concludes that the accuracy and reliability of the LLNA: BrdU-ELISA and 
LLNA: DA support their use to determine whether substances have the 
potential to cause ACD. The protocols also include reduced LLNA: BrdU-
ELISA and LLNA: DA procedures that should always be considered and used 
where determined appropriate because they can further reduce animal use 
by 40% compared to multi-dose procedures. The ICCVAM evaluation and 
complete recommendations for the LLNA: BrdU-ELISA and LLNA: DA are 
provided in the ICCVAM Test Method Evaluation Report on the Murine 
Local Lymph Node Assay: BrdU-ELISA, A Nonradioactive Alternative Test 
Method to Assess the Allergic Contact Dermatitis Potential of Chemicals 
and Products (NIH Publication No. 10-7552, available at http://iccvam.niehs.nih.gov/methods/immunotox/llna-ELISA/TMER.htm) and the 
ICCVAM Test Method Evaluation Report on the Murine Local Lymph Node 
Assay: DA, A Nonradioactive Alternative Test Method to Assess the 
Allergic Contact Dermatitis Potential of Chemicals and Products (NIH 
Publication No. 10-7551, available at http://iccvam.niehs.nih.gov/methods/immunotox/llna-DA/TMER.htm). The OECD subsequently adopted the 
LLNA: BrdU-ELISA and LLNA: DA as international test guidelines (OECD, 
2010b, 2010c).
    ICCVAM also concluded that available data support the use of the 
LLNA for safety testing of a broader range of chemicals and products, 
including pesticide formulations, metals with the exception of nickel, 
substances in aqueous solutions, and other chemicals and products, 
unless there are unique physicochemical properties associated with 
these materials that may interfere with the accuracy of the LLNA. 
Aqueous solutions should be tested in an appropriate vehicle that 
maintains sufficient contact of the test article with the skin. The 
ICCVAM evaluation and complete recommendations for expanded uses of the 
LLNA are provided in ICCVAM Test Method Evaluation Report on Using the 
Murine Local Lymph Node Assay for Testing Pesticide Formulations, 
Metals, Substances in Aqueous Solutions, and Other Products (NIH 
Publication No. 10-7512, available at http://iccvam.niehs.nih.gov/methods/immunotox/LLNA-app/TMER.htm).
    ICCVAM evaluated the new versions and applications of the LLNA in 
response to a 2007 nomination from CPSC (http://iccvam.niehs.nih.gov/methods/immunotox/llnadocs/CPSC_LLNA_nom.pdf). The nomination 
requested that ICCVAM assess (1) the validation status of the LLNA 
limit dose procedure (i.e., the reduced LLNA); (2) modified LLNA test 
method protocols that do not require the use of radioactive materials; 
(3) the use of the LLNA to test mixtures, aqueous solutions, and 
metals; and (4) the use of the LLNA as a stand-alone assay to determine 
ACD potency categories for hazard classification. ICCVAM 
recommendations on an updated LLNA test method protocol that included 
the reduced LLNA were communicated to Federal agencies and each of the 
15 ICCVAM agencies concurred with the ICCVAM recommendations for the 
reduced LLNA. ICCVAM has completed the evaluation of the LLNA for its 
validity for potency categorization of chemicals causing ACD in humans. 
Final ICCVAM recommendations will be forwarded to Federal agencies in 
2011.

Agency Responses to ICCVAM Recommendations

    In June 2010, ICCVAM forwarded final test method recommendations 
for the LLNA BrdU-ELISA, LLNA: DA and the expanded uses of the LLNA to 
U.S. Federal agencies for consideration (74 FR 50212), in accordance 
with the ICCVAM Authorization Act of 2000 (42 U.S.C. 285l-3(e)(4)). The 
Act requires agencies to review ICCVAM test method recommendations and 
notify ICCVAM in writing of their findings no later than 180 days after 
receipt of recommendations. The Act also requires ICCVAM to make ICCVAM 
recommendations and agency responses available to the public. Agency 
responses are to include identification of relevant test methods for 
which the ICCVAM test method recommendations may be added or 
substituted, and indicate any revisions or planned revisions to 
existing guidelines, guidances, or regulations to be made in response 
to these recommendations.
    Federal agency responses include acceptance decisions and agreement 
with the test method recommendations for the LLNA: BrdU-ELISA, LLNA: DA 
and the expanded uses of the LLNA. Several agencies also indicated that 
they would communicate the ICCVAM recommendations to stakeholders and 
encourage their appropriate use. Agency responses are available at 
http://iccvam.niehs.nih.gov/methods/immunotox/llna.htm.

[[Page 2390]]

Background Information on ICCVAM and NICEATM

    ICCVAM is an interagency committee composed of representatives from 
15 Federal regulatory and research agencies that require, use, 
generate, or disseminate toxicological and safety testing information. 
ICCVAM conducts technical evaluations of new, revised, and alternative 
methods with regulatory applicability and promotes the scientific 
validation and regulatory acceptance of toxicological and safety-
testing methods that more accurately assess the safety and hazards of 
chemicals and products and that reduce, refine (decrease or eliminate 
pain and distress), or replace animal use. The ICCVAM Authorization Act 
of 2000 established ICCVAM as a permanent interagency committee of the 
NIEHS under NICEATM (42 U.S.C. 285l-3(a)). NICEATM administers ICCVAM, 
provides scientific and operational support for ICCVAM-related 
activities, and conducts independent validation studies to assess the 
usefulness and limitation of new, revised, and alternative test 
methods. NICEATM and ICCVAM work collaboratively to evaluate new and 
improved test methods applicable to the needs of U.S. Federal agencies. 
NICEATM and ICCVAM welcome the public nomination of new, revised, and 
alternative test methods for validation studies and technical 
evaluations. Additional information about ICCVAM and NICEATM can be 
found on the NICEATM-ICCVAM Web site (http://iccvam.niehs.nih.gov).

References

EPA. 2003. Health Effects Test Guidelines: OPPTS 870.2600--Skin 
Sensitization. EPA 712-C-03-197. Washington, DC: U.S. Environmental 
Protection Agency. Available: http://www.epa.gov/ocspp/pubs/frs/publications/Test_Guidelines/series870.htm.
ICCVAM 2010 ICCVAM Test Method Evaluation Report. The Murine Local 
Lymph Node Assay: BrdU-ELISA, a Nonradioactive Alternative Test Method 
to Assess the Allergic Contact Dermatitis Potential of Chemicals and 
Products. NIH Publication No. 10-7552. Research Triangle Park, NC: 
National Institute of Environmental Health Sciences. Available at: 
http://iccvam.niehs.nih.gov/methods/immunotox/llna-ELISA/TMER.htm.
ICCVAM. 2010. ICCVAM Test Method Evaluation Report. The Murine Local 
Lymph Node Assay: DA, a Nonradioactive Alternative Test Method to 
Assess the Allergic Contact Dermatitis Potential of Chemicals and 
Products. NIH Publication No. 10-7551. Research Triangle Park, NC: 
National Institute of Environmental Health Sciences. Available at: 
http://iccvam.niehs.nih.gov/methods/immunotox/llna-D/TMER.htm.
ICCVAM. 2010. ICCVAM Test Method Evaluation Report on Using the LLNA 
for Testing Pesticide Formulations, Metals, Substances in Aqueous 
Solutions, and Other Products. NIH Publication No. 10-7512. Research 
Triangle Park, NC: National Institute of Environmental Health Sciences. 
Available at: http://iccvam.niehs.nih.gov/methods/immunotox/LLNA-app/TMER.htm.
ICCVAM. 2009. Recommended Performance Standards: Murine Local Lymph 
Node Assay. NIH Publication No. 09-7357. Research Triangle Park, NC: 
National Institute of Environmental Health Sciences. Available at: 
http://iccvam.niehs.nih.gov/methods/immunotox/llna_PerfStds.htm.
ICCVAM. 1999. The Murine Local Lymph Node Assay: A Test Method for 
Assessing the Allergic Contact Dermatitis Potential of Chemicals/
Compounds. The Results of an Independent Peer Review Evaluation 
Coordinated by ICCVAM and NICEATM. NIH Publication No. 99-4494. 
Research Triangle Park, NC: National Institute of Environmental Health 
Sciences. Available at: http://iccvam.niehs.nih.gov/methods/immunotox/llna_PeerPanel98.htm.
ISO. 2002. Biological evaluation of medical devices--10993 Part 10: 
Tests for irritation and delayed-type hypersensitivity. Available for 
purchase at: http://www.iso.org/iso/home.htm.
ISO. 2010. Biological evaluation of medical devices--10993 Part 10: 
Tests for irritation and skin sensitization. Available for purchase at: 
http://www.iso.org/iso/home.htm.
OECD. 2002. Test Guideline 429. Skin Sensitization: Local Lymph Node 
Assay, adopted April 24, 2002. In: OECD Guidelines for Testing of 
Chemicals. Paris: OECD.
OECD. 2010a. Test Guideline 429. Skin Sensitization: Local Lymph Node 
Assay, adopted July 22, 2010. In: OECD Guidelines for Testing of 
Chemicals. Paris: OECD. Available at: http://dx.doi.org/10.1787/9789264071100-en.
OECD. 2010b. Test Guideline 442B. Skin Sensitization: Local Lymph Node 
Assay: BrdU-ELISA, adopted July 22, 2010. In: OECD Guidelines for 
Testing of Chemicals. Paris: OECD. Available at: http://dx.doi.org/10.1787/9789264090996-en.
OECD. 2010c. Test Guideline 442A. Skin Sensitization: Local Lymph Node 
Assay: DA, adopted July 22, 2010. In: OECD Guidelines for Testing of 
Chemicals. Paris: OECD. Available at: http://dx.doi.org/10.1787/9789264090972-en.

    Dated: January 5, 2011.
John R. Bucher,
Associate Director, National Toxicology Program.
[FR Doc. 2011-669 Filed 1-12-11; 8:45 am]
BILLING CODE 4140-01-P