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The local lymph node assay (LLNA).

Costanza Rovida1, Cindy Ryan, Serena Cinelli

  • 1CAAT Europe, University of Konstanz, Konstanz, Germany.

Current Protocols in Toxicology
|April 19, 2012
PubMed
Summary
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Quantitative next generation risk assessment for skin sensitization - application of regression models based on in vitro data to estimate point of departure.

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The murine local lymph node assay (LLNA) assesses chemical skin sensitization. A new LLNA:BrdU-ELISA method replaces radioactive thymidine with 5-bromo-2-deoxyuridine (BrdU), improving animal welfare and safety.

Area of Science:

  • Toxicology
  • Dermatology
  • Immunotoxicology

Background:

  • The murine local lymph node assay (LLNA) is a standard in vivo method for evaluating chemical skin sensitization.
  • It offers significant animal welfare benefits over guinea pig tests, reducing animal numbers, pain, and trauma.
  • The LLNA is crucial for hazard identification and potency determination in risk assessment and is the only validated in vivo method.

Purpose of the Study:

  • To evaluate the LLNA:BrdU-ELISA method as a non-radioactive alternative for assessing skin sensitization.
  • To measure lymph node cell (LNC) proliferation using 5-bromo-2-deoxyuridine (BrdU) incorporation.

Main Methods:

  • The study focuses on the LLNA:BrdU-ELISA variant of the local lymph node assay.
  • This method measures LNC proliferation in draining lymph nodes.

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  • It utilizes 5-bromo-2-deoxyuridine (BrdU) as a substitute for radioactive thymidine.

    • Main Results:

    • The LLNA:BrdU-ELISA method provides a non-radioactive means to assess LNC proliferation.
    • This variant allows for hazard identification and potency determination of contact allergens.
    • It maintains the advantages of the original LLNA regarding animal welfare.

    Conclusions:

    • The LLNA:BrdU-ELISA is a viable and validated alternative to the radiolabeled thymidine method.
    • This non-radioactive approach enhances safety and animal welfare in skin sensitization testing.
    • It supports robust risk assessment of chemical sensitizers.