[Federal Register Volume 75, Number 124 (Tuesday, June 29, 2010)]
[Notices]
[Pages 37443-37445]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 2010-15777]



[[Page 37443]]

=======================================================================
-----------------------------------------------------------------------

DEPARTMENT OF HEALTH AND HUMAN SERVICES


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

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

ACTION: Availability of Reports; Notice of Transmittal.

-----------------------------------------------------------------------

SUMMARY: NICEATM announces availability of Interagency Coordinating 
Committee on the Validation of Alternative Methods (ICCVAM) Test Method 
Evaluation Reports (TMERs) recommending two nonradioactive versions of 
the 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. Related ICCVAM Test Method Recommendations in each report 
have also been transmitted to Federal agencies for their review and 
response to ICCVAM in accordance with the provisions of the ICCVAM 
Authorization Act of 2000. The LLNA: 5-Bromo-2'-deoxyuridine-Enzyme-
Linked Immunosorbent Assay (BrdU-ELISA) and LLNA: Daicel Adenosine 
Triphosphate (DA) do not use radioactive reagents and therefore provide 
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 use of these test methods to 
identify substances as potential skin sensitizers or nonsensitizers. 
Based on an updated evaluation, ICCVAM is also recommending expanded 
use of the LLNA to evaluate the ACD hazard potential of pesticide 
formulations and other products.

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

    ICCVAM previously evaluated the validation status of the LLNA as a 
stand-alone alternative method to the guinea pig maximization test 
(GPMT) and the Buehler test (BT) for assessing the ACD hazard potential 
of products and chemicals (NIH Publication No. 99-4494; available at 
http://iccvam.niehs.nih.gov/methods/immunotox/llna.htm). Based on this 
evaluation, ICCVAM recommended the LLNA as a valid substitute for the 
guinea pig test methods for most testing situations in 1999. The 
Environmental Protection Agency, the 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. The 
Organization for Economic Co-operation and Development (OECD) 
subsequently adopted the LLNA as OECD Test Guideline 429 in 2002. Using 
the LLNA instead of guinea pig tests reduces and refines (less pain and 
distress) animal use for ACD safety testing.
    In 2007, the CPSC nominated several new versions and applications 
of the LLNA to ICCVAM for evaluation of their scientific validity 
(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) the 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 to 
determine ACD potency categories for hazard classification. NICEATM 
published a Federal Register notice (72 FR 27815) requesting public 
comments on (1) the appropriateness and relative priority of the CPSC-
nominated LLNA activities, (2) the nomination of scientists to serve on 
an international independent scientific peer review panel, and (3) the 
submission of data from LLNA testing that related to the CPSC-nominated 
LLNA activities as well as corresponding data from human and other 
animal studies. ICCVAM assigned these activities a high priority after 
considering comments from the public and endorsement from the 
Scientific Advisory Committee on Alternative Toxicological Methods 
(SACATM). NICEATM and ICCVAM compiled comprehensive draft background 
review documents (BRDs), released them for public comment in January 
2008 (73 FR 1360), and convened a public meeting of the panel on March 
4-6, 2008 to peer review the draft documents. The panel evaluated the 
information in the BRDs as to whether it supported draft ICCVAM test 
method recommendations for test method uses and limitations, updated 
standardized test method protocols, and proposed future studies. The 
panel considered public comments made at the meeting, as well as public 
comments submitted in advance of the meeting, before concluding their 
deliberations. The panel's report was made available in May 2008 (73 FR 
29136) for public comment. The draft ICCVAM BRDs, draft ICCVAM test 
method recommendations, the panel's report, and all public comments 
were made available to SACATM for comment at its meeting on June 18-19, 
2008 (73 FR 25754).
    After considering the conclusions and recommendations of the panel, 
comments from SACATM, and public comments, ICCVAM forwarded final 
recommendations for the updated LLNA test method protocol, the reduced 
LLNA, and LLNA performance standards to Federal agencies in September 
2009 (74 FR 50212). ICCVAM concluded that the updated LLNA test method 
protocol will further reduce animal use by 20% compared to the original 
version of the LLNA and also provide for more consistent and reliable 
results. The reduced LLNA will reduce animal use by 40% for each test 
compared to the traditional, multi-dose LLNA. ICCVAM also recommended 
LLNA test method performance standards that can be used to efficiently 
evaluate the validity of modified versions of the LLNA that are 
mechanistically and functionally similar to the traditional LLNA. 
Federal agencies subsequently responded with their support and 
concurrence with the ICCVAM recommendations. Agency responses are 
available on the NICEATM-ICCVAM Web site.
    NICEATM subsequently obtained additional data and/or information 
and revised the draft documents for both the traditional and 
nonradioactive LLNA methods. ICCVAM released the revised draft 
documents to the public for comment and announced a second meeting of 
the panel (74 FR 8974). The panel reconvened in public session on April 
28-29, 2009 to review the ICCVAM-revised draft documents and

[[Page 37444]]

finalize its conclusions and recommendations on the current validation 
status of the nonradioactive test methods and the expanded uses of the 
LLNA for pesticide formulations and other products. The panel's report 
was made available for public comment in June 2009 (74 FR 26242). The 
revised draft ICCVAM BRDs, revised draft ICCVAM test method 
recommendations, the panel's report, and all public comments were made 
available to SACATM for comment on June 25-26, 2009 (74 FR 19562). 
After considering the conclusions and recommendations of the panel, 
comments from SACATM, and public comments, along with the 
recommendations of an OECD Expert Consultation on the LLNA convened in 
October and December 2009, ICCVAM finalized and forwarded test method 
recommendations to Federal agencies for their consideration, in 
accordance with the ICCVAM Authorization Act of 2000 (42 U.S.C. 285l-
3(e)(4)). Agency responses to the ICCVAM test method recommendations 
will be made available on the NICEATM-ICCVAM website as they are 
received.
    The ICCVAM TMERs, The LLNA: BrdU-ELISA, A Nonradioactive 
Alternative Test Method to Assess the Allergic Contact Dermatitis 
Potential of Chemicals and Products (NIH Publication 10-7552), and The 
LLNA: DA, A Nonradioactive Alternative Test Method to Assess the 
Allergic Contact Dermatitis Potential of Chemicals and Products (NIH 
Publication 10-7551), describe ICCVAM's recommendations for using the 
LLNA: BrdU-ELISA and LLNA: DA for regulatory hazard identification 
purposes. The reports also provide ICCVAM-recommended LLNA: BrdU-ELISA 
and LLNA: DA test method protocols, the final BRDs, and the peer review 
reports of the panel. The ICCVAM-recommended LLNA: BrdU-ELISA test 
method protocol is based on the protocol developed by Takeyoshi et al. 
(2001). The ICCVAM-recommended LLNA: DA test method protocol is based 
on the protocol developed by Idehara et al. (2008). Both test method 
protocols incorporate all relevant aspects of the recently updated 
ICCVAM-recommended traditional LLNA test method protocol (ICCVAM, 
2009). The protocols also include reduced LLNA: BrdU-ELISA and LLNA: DA 
procedures that should always be considered and used where determined 
appropriate in order to further reduce animal use.
    The ICCVAM Test Method Evaluation Report, Using the Murine Local 
Lymph Node Assay for Testing Pesticide Formulations, Metals, Substances 
in Aqueous Solutions, and Other Products (NIH Publication 10-7512) 
provides ICCVAM's updated evaluation and recommendations for use of the 
LLNA to evaluate the ACD hazard potential of pesticide formulations, 
metals, substances in aqueous solutions, and other products. The 
evaluation considered new data that became available subsequent to the 
original ICCVAM LLNA evaluation in 1999. The report also includes the 
peer review reports of the panel.
    ICCVAM's evaluation of the LLNA for skin sensitization potency 
categorization is currently nearing completion, and final ICCVAM 
recommendations will be forwarded to Federal agencies later this year.

Background Information on ICCVAM, NICEATM, and SACATM

    ICCVAM is an interagency committee composed of representatives from 
15 Federal regulatory and research agencies that require, use, 
generate, or disseminate toxicological information. ICCVAM conducts 
technical evaluations of new, revised, and alternative methods with 
regulatory applicability and promotes the scientific validation and 
regulatory acceptance of toxicological test methods that more 
accurately assess the safety and hazards of chemicals and products that 
refine, reduce, and 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). NICEATM administers ICCVAM and 
provides scientific and operational support for ICCVAM-related 
activities. 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 both validation studies as well as 
technical evaluations. Additional information about ICCVAM and NICEATM 
can be found on the NICEATM-ICCVAM Web site (http://iccvam.niehs.nih.gov).
    SACATM was established January 9, 2002, and is composed of 
scientists from the public and private sectors (67 FR 11358). SACATM 
provides advice to the Director of the NIEHS, ICCVAM, and NICEATM 
regarding the statutorily-mandated duties of ICCVAM and activities of 
NICEATM. Additional information about SACATM, including the charter, 
roster, and records of past meetings, can be found at http://ntp.niehs.nih.gov/go/167.

References

    ICCVAM. 2010. ICCVAM Test Method Evaluation Report on The LLNA: 
BrdU-ELISA, A Nonradioactive Alternative Test Method to Assess the 
Allergic Contact Dermatitis Potential of Chemicals and Products. NIH 
Publication Number 10-7552. Research Triangle Park, NC: National 
Institute of Environmental Health Sciences.
    ICCVAM. 2010. ICCVAM Test Method Evaluation Report on The LLNA: DA, 
A Nonradioactive Alternative Test Method to Assess the Allergic Contact 
Dermatitis Potential of Chemicals and Products. NIH Publication Number 
10-7551. Research Triangle Park, NC: National Institute of 
Environmental Health Sciences.
    ICCVAM. 2010. 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 Number 10-7512. Research Triangle Park, NC: National 
Institute of Environmental Health Sciences.
    ICCVAM. 2009. Recommended Performance Standards: Murine Local Lymph 
Node Assay. NIH Publication Number 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. 2009. ICCVAM Test Method Evaluation Report. The Reduced 
Murine Local Lymph Node Assay: An Alternative Test Method Using Fewer 
Animals to Assess the Allergic Contact Dermatitis Potential of 
Chemicals and Products. NIH Publication Number 09-6439. Research 
Triangle Park, NC: National Institute of Environmental Health Sciences. 
Available at: http://iccvam.niehs.nih.gov/methods/immunotox/LLNA-LD/TMER.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 Number 99-4494. 
Research Triangle Park, NC: National Institute of Environmental Health 
Sciences. Available at: http://iccvam.niehs.nih.gov/methods/immunotox/llna.htm.
    Idehara K, Yamagishi G, Yamashita K, Ito M. 2008. Characterization 
and evaluation of a modified local lymph node assay using ATP content 
as a non-radio isotopic endpoint. Journal of Pharmacological and 
Toxicological Methods 58(1): 1-10.
    Takeyoshi M, Yamasaki K, Yakabe Y, Takatsuki M, Kimber I. 2001. 
Development of non-radio isotopic endpoint of murine local lymph node

[[Page 37445]]

assay based on 5-bromo-2'-deoxyuridine (BrdU) incorporation. Toxicology 
Letters 119(3): 203-208.

    Dated: June 16, 2010.
John R. Bucher,
Associate Director, National Toxicology Program.
[FR Doc. 2010-15777 Filed 6-28-10; 8:45 am]
BILLING CODE 4140-01-P