Murine Local Lymph Node Assay | EURL ECVAM

Topic: Sensitisation

Test Method Number:

TM1998-05 (US)

Short Name of TM:

Murine Local Lymph Node Assay

Year received:

1998

Responsible Organisation:

ICCVAM/NICEATM - United States

General Comments:

Method Description

Skin sensitizers induce a primary sensitization of lymphocytes in the auricular lymph nodes that drain the site of test chemical application. In the LLNA, lymphocyte proliferation is proportional to the dose of test chemical applied and is used as a measure of sensitization after weighing the lymph nodes, then extracting and labeling the cells with 3H-methyl thymidine and measuring disintegrations per minute (DPM).

Track Approval Status

Submission
Validation
Peer-review
Recommendation
Regulatory acceptance/Standards

Submission

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Date	Stage	Comment	Documents/Links
Apr 1998	Assessment finalised	ICCVAM evaluated an LLNA submission from Proctor & Gamble that included LLNA publications and the results of several European validation studies.

Validation

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Date	Stage	Comment	Documents/Links
Apr 1998	Finalised	ICCVAM conducted a review of LLNA performance in the Proctor & Gamble submission containing published literature and results of European validation studies and recommended it as an alternative to guinea pig tests for assessing the allergic potential of chemicals to cause ACD.
Mar 2009	Finalised	ICCVAM, in conjunction with ECVAM and JaCVam, developed harmonized performance standards (PS) for the LLNA, based on performance of the LLNA compared to results from guinea pig tests and human outcomes, that could be used to evaluate modified versions of the LLNA. The PS's included essential test method components, a miminum list of reference substances, and standards for accuracy and reliability.

Peer-review

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Date	Stage	Comment	Documents/Links
Mar 1999	Finalised	ICCVAM coordinated an independent peer review evaluation of the LLNA who concluded that the LLNA was equivalent to the current guinea pig methods and offered several advantages including refinement of eliminating pain and distress in animals compared to the GPMT and opportunities for future assay improvement and mechanistic studies.	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 bu ICCVAM and NICEATM

Recommendation

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Date	Stage	Comment	Documents/Links
Mar 2010	Published	ICCVAM/NICEATM published a test method evaluation report on the murine LLNA: BrdU-ELISA, A nonradioactive alternative test method to assess the ACD potential of chemicals and products and concluded that the accuracy and reliability of the test method support its use to identify substances as potential skin sensitizers or nonsensitizers.	ICCVAM Test Method Evaluation Report on the Murine LLNA:BrdU-ELISA, a Nonradioactive Alternative Test Method to Assess the ACD Potential of Chemicals and Products
Mar 2010	Published	ICCVAM/NICEATM published a Test Method Evaluation Report on the Murine LLNA: DA (test method developed by Daicel Chemical Industries, Ltd. in Hyogo, Japan that measures ATP content as an indicator of lymphocyte proliferation), a Nonradioactive Alternative Test Method to Assess the ACD Potential of Chemicals and Products and concluded that the accuracy and reliability of the test method support its use to identify potential skin sensitizers and nonsensitizers.	ICCVAM Test Method Evaluation Report on the Murine LLLNA: DA, a Nonradioactive Test Method to Assess the ACD Potential of Chemicals and Products
Jun 2011	Published	ICCVAM evaluated the usefulness and limitations of the murine LLNA for potency categorization of chemicals causing allergic contact dermatitis in humans and concluded that the LLNA can be used to categorize substances as strong sensitizers using the GHS Subcategory 1A, when the estimated concentration that produces a positive LLNA result (i.e., EC3) is <=2%, but additional information is needed for Subcategory 1B, when the substance produces an LLNA EC3 >2%.	ICCVAM Test Method Evaluation Report: Usefulness and Limitations of the Murine LLNA for Potency Categorization of Chemicals Causing ACD in Humans

Regulatory acceptance/Standards

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Date	Stage	Comment	Documents/Links
Jul 2010	Revision of existing regulatory standard/guideline		OECD TG 429: Skin Sensitization: Local Lymph Node Assay
Jun 2018	Adopted/published		OECD TG 442B: Local lymph node assay: BRDU-ELISA or FCM