Test Method Evaluation Report: Nonradioactive LLNA:DA

Cover of the ICCVAM Test Method Evaluation Report on the LLNA:DAICCVAM 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

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Complete Report

Front Matter

Includes Table of Contents, Lists of Tables and Figures, List of Abbreviations and Acronyms, Acknowledgements, Preface, and Executive Summary

Main Body

  • Section 1: Introduction
  • Section 2: ICCVAM Recommendations for the Nonradioactive LLNA: DA Test Method
  • Section 3: Validation Status of the LLNA: DA Test Method
  • Section 4: ICCVAM Consideration of Independent Peer Review Panel Report and Other Comments

Appendix A: ICCVAM Evaluation Timeline

Appendix B: ICCVAM-Recommended Protocol: The Murine Local Lymph Node Assay: DA, a Nonradioactive Alternative Test Method to Assess the Allergic Contact Dermatitis Potential of Chemicals and Products

  • Main Body
  • Annex I: An Approach to Dissection and Identification of the Draining (“Auricular”) Lymph Nodes
  • Annex II: An Example of How to Reduce the Number of Animals in the Concurrent Positive Control Group of the Local Lymph Node Assay
  • Annex III: Evaluating Local Irritation and Systemic Toxicity in the Local Lymph Node Assay

Appendix C: Final Background Review Document: The Nonradioactive Murine Local Lymph Node Assay: DA 

  • Main Body
    • Front matter: Includes Table of Contents, Acknowledgements, Preface, and Executive Summary
    • Section 1: Introduction
    • Section 2: LLNA: DA Test Method Protocol
    • Section 3: LLNA: DA Validation Database
    • Section 4: Reference Data
    • Section 5: LLNA: DA Test Method Data and Results
    • Section 6: LLNA: DA Test Method Accuracy
    • Section 7: LLNA: DA Test Method Reliability
    • Section 8: LLNA: DA Data Quality
    • Section 9: Other Scientific Reports and Reviews
    • Section 10: Animal Welfare Considerations
    • Section 11: Practical Considerations
    • Section 12: References
    • Section 13: Glossary
  • Annex I: LLNA: DA Test Method Protocol
  • Annex II: Physicochemical Properties and Chemical Classes of Substances Tested in the LLNA: DA
  • Annex III: Comparative LLNA: DA, Traditional LLNA, Guinea Pig, and Human Skin Sensitization Data
  • Annex IV: Data for the LLNA: DA Intralaboratory and Interlaboratory Validation Studies
  • Annex V: Accuracy Analyses Using Additional Approaches for Combining Multiple Test Results
  • Annex VI: Evaluation of the Robustness of the SI Cutoff Criteria Used for the LLNA: BrdU-ELISA and LLNA: DA Test Methods
  • Annex VII: Analyses Using Multiple SI Decision Criteria
  • Annex VIII: Reproducibility Analyses for the LLNA: DA Using a Single Decision Criterion of SI ≥ 3.0 or SI ≥ 2.0

Appendix D: Independent Scientific Peer Review Panel Assessment

  • Appendix D1: Summary Minutes from Peer Review Panel Meeting on March 4-6, 2008
  • Appendix D2: Independent Scientific Peer Review Panel Report: Validation Status of New Versions and Applications of the Murine Local Lymph Node Assay: A Test Method for Assessing the Allergic Contact Dermatitis Potential of Chemicals and Products
  • Appendix D3: Summary Minutes of Independent Scientific Peer Review Panel Meeting on April 28-29, 2009
  • Appendix D4: Independent Scientific Peer Review Panel Report: Updated Validation Status of New Versions and Applications of the Murine Local Lymph Node Assay: A Test Method for Assessing the Allergic Contact Dermatitis Potential of Chemicals and Products

Appendix E: JaCVAM Statement on the LLNA: DA for Skin Sensitization Testing

Appendix F: Federal Register Notices and Public Comments

  • Appendix F1: Federal Register Notices
    • 72 FR 27815 (May 17, 2007): The Murine Local Lymph Node Assay: Request for Comments, Nominations of Scientific Experts, and Submission of Data
    • 72 FR 52130 (September 12, 2007): Draft Performance Standards for the Murine Local Lymph Node Assay: Request for Comments
    • 73 FR 1360 (January 8, 2008): Announcement of an Independent Scientific Peer Review Panel Meeting on the Murine Local Lymph Node Assay; Availability of Draft Background Review Documents; Request for Comments
    • 73 FR 25754 (May 7, 2008): Meeting of the Scientific Advisory Committee on Alternative Toxicological Methods (SACATM)
    • 73 FR 29136 (May 20, 2008): Peer Review Panel Report on the Validation Status of New Versions and Applications of the Murine Local Lymph Node Assay (LLNA): A Test Method for Assessing the Allergic Contact Dermatitis Potential of Chemicals and Products: Notice of Availability and Request for Public Comments
    • 74 FR 8974 (February 27, 2009): Announcement of a Second Meeting of the Independent Scientific Peer Review Panel on the Murine Local Lymph Node Assay; Availability of Draft Background Review Documents (BRD); Request for Comments
    • 74 FR 19562 (April 29, 2009): Meeting of the Scientific Advisory Committee on Alternative Toxicological Methods (SACATM)
    • 74 FR 26242 (June 1, 2009): Independent Scientific Peer Review Panel Report: Updated Validation Status of New Versions and Applications of the Murine Local Lymph Node Assay: A Test Method for Assessing the Allergic Contact Dermatitis Potential of Chemicals and Products: Notice of Availability and Request for Public Comments
  • Appendix F2: Public Comments Received in Response to Federal Register Notices
  • Appendix F3: SACATM Comments: SACATM Meeting on June 18-19, 2008
    Please contact NICEATM to obtain this document
  • Appendix F4: SACATM Comments: SACATM Meeting on June 25-26, 2009
    Download minutes of the SACATM meeting from the NTP website

Appendix G: Relevant Skin Sensitization Regulations and Testing Guidelines

  • Appendix G1: Table of Relevant Skin Sensitization Test Regulations
  • Appendix G2: Health Effects Test Guidelines OPPTS 870.2600: Skin Sensitization
  • Appendix G3: ISO 10993-10: Biological Evaluation of Medical Devices Part 10: Tests for Irritation and Delayed-type Hypersensitivity
    Available for purchase on the ISO website
  • Appendix G4: OECD Guideline 429: Skin Sensitisation – Local Lymph Node Assay
  • Appendix G5: OECD Guideline 406: Skin Sensitisation