National Toxicology Program

References

Relevant Test Method Guidelines

EPA. 2003. Health Effects Test Guidelines OPPTS 870.2600 Skin Sensitization. EPA 712-C-03-197. Washington, DC: U.S. Environmental Protection Agency. [PDF]

OECD. 2002. Guideline For Testing of Chemicals, 429, Skin Sensitisation: Local Lymph Node Assay. Paris, France: Organisation for Economic Co-operation and Development. [PDF]

Background on the LLNA

Sailstad DM. 2002. Murine local lymph node assay: an alternative test method for skin hypersensitivity testing. Lab Animal 31:36-41.

Sailstad D, Hattan D, Hill R, and Stokes WS. 2001. Evaluation of the murine local lymph node assay (LLNA) I: The ICCVAM review process. Regulatory Toxicology and Pharmacology 34:249-257.

Dean J, Twerdok L, Tice R, Sailstad D, Hattan D and Stokes, WS. 2001. Evaluation of the murine local lymph node assay (LLNA) II: Conclusions and recommendations of an independent scientific peer review panel. Regulatory Toxicology and Pharmacology 34:258-273

Haneke K, Tice R, Carson B, Margolin B, and Stokes WS. 2001. Evaluation of the murine local lymph node assay (LLNA) III: Data analyses completed by the National Toxicology Program (NTP) Interagency Center for the Evaluation of Alternative Toxicological Methods (NICEATM). Regulatory Toxicology and Pharmacology 34:274-286.

Cut-Down Assay

I Kimber, R Dearman, C Betts, G Gerberick, C Ryan, P Kern, et al. 2006. The local lymph node assay and skin sensitization: a cut-down screen to reduce animal requirements? Contact Dermatitis 54:181-185.

LLNA Use to Assess Sensitization Potency

D Basketter, J McFadden, P Evans, K Andersen, I Jowsey. 2006. Identification and classification of skin sensitizers: identifying false positives and false negatives. Contact Dermatitis 55:268-273.

D Basketter, I Kimber. 2006. Predictive test for irritants and allergens and their use in quantitative risk assessment. In: Contact Dermatitis, 4th Edition (P Frosch, T Menn, J-P Lepoittevin, eds.) Heidelberg: Springer Verlag, 179–188.

D Basketter, K Andersen, C Carola, H Van Loveren, A Boman, I Kimber, et al. 2005. Evaluation of the skin sensitizing potency of chemicals by using the existing methods and considerations of relevance for elicitation. Contact Dermatitis 52:39-43.

D Basketter, C Clapp, D Jefferies, R Safford, C Ryan, G Gerberick, R Dearman, I Kimber. 2005. Predictive identification of human skin sensitisation thresholds. Contact Dermatitis 53:260-267.

D Basketter, P Cadby. 2004. Reproducible prediction of contact allergenic potency using the local lymph node assay. Contact Dermatitis 50:15-17.

D Basketter, C Smith Pease, G Patlewicz. 2003. Contact allergy: the local lymph node assay for the prediction of hazard and risk. Clinical and Experimental Dermatology 28:218-221.

D Basketter, L Blaikie, R Dearman, I Kimber, C Ryan, G Gerberick, P Harvey, P Evans, I White, R Rycroft. 2000. Use of the local lymph node assay for the estimation of relative contact allergenic potency. Contact Dermatitis 42:344-348.

D Basketter, L Lea, K Cooper, A Dickens, D Briggs, I Pate, R Dearman, I Kimber. 1999. A comparison of statistical approaches to derivation of EC3 values from local lymph node assay dose responses. Journal of Applied Toxicology 19:261-266.

D Basketter, L Lea, K Cooper, A Dickens, J Stocks, I Pate, R Dearman, I Kimber. 1999. Thresholds for classification as a skin sensitiser in the local lymph node assay: a statistical evaluation. Food and Chemical Toxicology 37:1167-1174.

D Basketter, R Rodford, I Kimber, I Smith, J Wahlberg. 1999. Skin sensitization risk assessment: a comparative evaluation of three isothiazolinone biocides. Contact Dermatitis 40:150-154.

D Basketter, D Roberts, M Cronin, E Scholes. 1992. The value of the local lymph node assay in quantitative structure activity investigations. Contact Dermatitis 27:137-142.

D Basketter, Z Wright, E Warbrick, R Dearman, I Kimber, C Ryan, G Gerberick, I White. 2001. Human potency predictions for aldehydes using the local lymph node assay. Contact Dermatitis 45:89-94.

C Betts, R Dearman, J Heylings, I Kimber, D Basketter. 2006. Skin sensitization potency of methyl methacrylate in the local lymph node assay: comparisons with guinea-pig data and human experience. Contact Dermatitis 55:140-147.

C Betts, R Dearman, I Kimber, H Maibach. 2005. Potency and risk assessment of a skin-sensitizing disperse dye using the local lymph node assay. Contact Dermatitis 52:268-272.

R Dearman, Z Wright, D Basketter, C Ryan, G Gerberick, I Kimber. 2001. The suitability of hexyl cinnamic aldehyde as a calibrant for the using local lymph node assay. Contact Dermatitis 44:357-361.

G Gerberick, C Ryan, R Dearman, I Kimber. 2007. Local lymph node assay (LLNA) for detection of sensitization capacity of chemicals. Methods 41:54-60.

G Gerberick, C Ryan, P Kern, H Schlatter, R Dearman, I Kimber, G Patlewicz, D Basketter. 2007. Updating the compilation of historical local lymph node assay data for the evaluation of skin sensitization alternatives. Dermatitis: MS in preparation.

G Gerberick, C Ryan, P Kern, H Schlatter, R Dearman, I Kimber, G Patlewicz, D Basketter. 2005. Compilation of historical local lymph node assay data for the evaluation of skin sensitization alternatives. Dermatitis 16:157-202.

G Gerberick, Robinson MK, C Ryan, R Dearman, I Kimber, D Basketter, Wright Z, Marks JG. 2001. Contact allergenic potency: correlation of human and local lymph node assay data. American Journal of Contact Dermatitis 12:156-161.

G Gerberick, J Vassallo, L Foertsch, B Price, J Chaney, J-P Lepoittevin. 2007. Quantification of chemical peptide reactivity for screening contact allergens: a classification tree model approach. Toxicological Sciences 97:417-427.

P Griem, C Goebel, H Scheffler. 2003. Proposal for a risk assessment methodology for skin sensitization based on sensitization potency data. Regulatory Toxicology and Pharmacology 38:269-290.

J Hilton, R Dearman, P Harvey, P Evans, D Basketter, I Kimber. 1998. Estimation of the relative skin sensitizing potency using the local lymph node assay: a comparison of formaldehyde with glutaraldehyde. American Journal of Contact Dermatitis 9:29-33.

I Jowsey, D Basketter, C Westmoreland, I Kimber. 2006. A future approach to measuring relative skin sensitising potency: a proposal. Journal of Applied Toxicology 26:341-350.

I Kimber, D Basketter, M Butler, A Gamer, J Garrigue, G Gerberick, et al. 2003. Classification of contact allergens according to potency: proposals. Food and Chemical Toxicology 41:1799-1809.

I Kimber, D Basketter, K Berthold, M Butler, J Garrigue, L Lea, C Newsome, R Roggeband, W Steiling, G Stropp, S Waterman, C Wiemann. 2001. Skin sensitization testing in potency and risk assessment. Toxicological Sciences 59:198-208.

I Kimber, G Gerberick, D Basketter. 1999. Thresholds in contact sensitization: theoretical considerations and practical applications. Food and Chemical Toxicology 37:553-560.

I Kimber, D Basketter. 1997. Contact sensitization: a new approach to risk assessment. Human and Ecological Risk Assessment (HERA) 3:385-395.

I Kimber, J Hilton, R Dearman, G Gerberick, C Ryan, D Basketter, E Scholes, R House, A Guy, G Ladics, S Loveless. 1995. An international evaluation of the murine local lymph node assay and comparison of modified procedures. Toxicology 103:63-73.

I Kimber, R Dearman. 1991. Investigation of lymph node cell proliferation as a possible immunological correlate of contact sensitizing potential. Food and Chemical Toxicology 29:125–129.

J Lalko, A Api. 2006. Investigation of the dermal sensitization potential of various essential oils in the local lymph node assay. Food and Chemical Toxicology 44:739-746.

L Lea, V Warbrick, R Dearman, P Harvey, I Kimber, D Basketter. 1999. The impact of vehicle on assessment of relative skin sensitisation potency of 1,4-dihydroquinone in the local lymph node assay. American Journal of Contact Dermatitis 10:213–218.

S Loveless, G Ladics, G Gerberick, C Ryan, D Basketter, E Scholes, R House, J Hilton, R Dearman, I Kimber. 1996. Further evaluation of the local lymph node assay in the final phase of an international collaborative trial. Toxicology 108:141-152.

H McGarry. 2007. The murine local lymph node assay: Regulatory and potency considerations under REACH. Toxicology 238:71–89.

G Patlewicz, D Basketter, C Smith, S Hotchkiss, D Roberts. 2001. Skin sensitisation structure activity relationships for aldehydes. Contact Dermatitis 44:331-336.

C Ryan, J Chaney, P Kern, R Dearman, I Kimber, D Basketter, G Gerberick. 2007. Extrapolating local lymph node assay EC3 values to estimate relative sensitizing potency. Journal of Toxicology: Cutaneous and Ocular Toxicology 26:135-45.

C Ryan, G Gerberick, L Cruse, D Basketter, L Lea, L Blaikie, R Dearman, E Warbrick, I Kimber. 2000. Activity of human contact allergens in the murine local lymph node assay. Contact Dermatitis 43:95-102.

E Schlede, W Aberer, T Fuchs, I Gerner, H Lessmann, T Maurer, et al. 2003. Chemical substances and contact allergy - 244 Substances ranked according to allergenic potency. Toxicology 193:219-259.

K Schneider, Z Akkan. 2004. Quantitative relationship between the local lymph node assay and human skin sensitization assays. Regulatory Toxicology and Pharmacology 39:245-255.

E Warbrick, R Dearman, L Lea, D Basketter, I Kimber. 1999. Local lymph node assay responses to paraphenylenediamine: intra- and inter-laboratory evaluations. Journal of Applied Toxicology 19:255-260.

Non-Radioactive LLNA Methods

P Azam, J Peiffer, J Ourlin , P Bonnet, M Tissier, L Vian, et al. 2005. Qualitative and quantitative evaluation of a local lymph node assay based on ex vivo interleukin-2 production. Toxicology 206:285-298.

G Gerberick, L Cruse, C Ryan. 1999. Local lymph node assay: differentiating allergic and irritant responses using flow cytometry. Methods: A Companion to Methods in Enzymology 19:48-55.

V Hans-Werner, A Jurgen. 2005. The local lymph node assay being too sensitive? Archives of Toxicology 79:721-728.

T Hariya, M Hatao, H Ichikawa. 1999. Development of a non-radioactive endpoint in a modified local lymph node assay. Food and Chemical Toxicology 37:87-93.

M Hatao, T Hariya, Y Katsumura, S Kato. 1995. A modification of the local lymph node assay for contact allergenicity screening: measurement of interleukin-2 as an alternative to radioisotope-dependent proliferation assay. Toxicology 98:15-22.

N Humphreys, R Dearman, I Kimber. 2003. Assessment of cumulative allergen-activated lymph node cell proliferation using flow cytometry. Toxicological Sciences 73:80-89.

U Kuhn, U Lempertz, J Knop, D Becker. 1995. A new method for phenotyping proliferating cell nuclear antigen positive cells using flow cytometry: implications for analysis of the immune response in vivo. Journal of Immunological Methods 179:215-222.

J Lee, J Park, S Park, H Kim, H Oh. 2002. A nonradioisotopic endpoint for measurement of lymph node cell proliferation in a murine allergic contact dermatitis model, using bromodeoxyuridine immunohistochemistry. Journal of Pharmacological and Toxicological Methods 48(1):53-61.

A Suda, M Yamashita, M Tabei, K Taguchi, H Vohr, N Tsutsui, et al. 2002. Local lymph node assay with non-radioisotope alternative endpoints. Journal of Toxicological Sciences 27:205-218.

M Takeyoshi, S Noda, K Yamasaki, I Kimber. 2006. Advantage of using CBA/N strain mice in a non-radioisotopic modification of the local lymph node assay. Journal of Applied Toxicology 26:5-9.

M Takeyoshi, M Sawaki, K Yamasaki, I Kimber. 2003. Assessment of statistic analysis in non-radioisotopic local lymph node assay (non-RI-LLNA) with α-hexylcinnamic aldehyde as an example. Toxicology 191:259-263.

M Takeyoshi, K Yamasaki, Y Yakabe, M Takatsuki, I Kimber. 2001. Development of non-radio isotopic endpoint of murine local lymph node assay based on 5-bromo-2'-deoxyuridine (BrdU) incorporation. Toxicology Letters 119:203-208.

G Ehling, M Hecht, A Heusener, J Huesler, A Gamer, H van Loveren, T Maurer, K Riecke, L Ullmann, P Ulrich, R Vandebriel, H Vohr. 2005. An European inter-laboratory validation of alternative endpoints of the murine local lymph node assay: first round. Toxicology 212:60-68.

G Ehling, M Hecht, A Heusener, J Huesler, A Gamer, H van Loveren, T Maurer, K Riecke, L Ullmann, P Ulrich, R Vandebriel, H Vohr. 2005. An European inter-laboratory validation of alternative endpoints of the murine local lymph node assay: 2nd round. Toxicology. 212:69-79.

LLNA Use in Testing of Mixtures, Formulations, Metals or Aqueous Solutions

F Andersen. 2005. Final report on the safety assessment of potassium silicate, sodium metasilicate, and sodium silicate. International Journal of Toxicology 24(SUPPL. 1):103-117.

D Basketter, L Lea, K Cooper, C Ryan, G Gerberick, R Dearman, et al. 1999. Identification of metal allergens in the local lymph node assay. American Journal of Contact Dermatitis 10:207-212.

F Clottens, A Breyssens, H De Raeve, M Demedts, B Nemery. 1996. Assessment of the ear swelling test and the local lymph node assay in hamsters. Journal of Pharmacological and Toxicological Methods 35:167-172.

T Hariya, M Hatao, H Ichikawa. 1999. Development of a non-radioactive endpoint in a modified local lymph node assay. Food and Chemical Toxicology 37:87-93.

J Hostynek, H Maibach. 2004. Copper hypersensitivity: dermatologic aspects. Dermatologic therapy 17:328-333.

--- 2003. Copper Hypersensitivity: Dermatologic aspects - an overview. Reviews on Environmental Health 18:153-183.

Y Ikarashi, M Kaniwa, T Tsuchiya. 2002. Sensitization potential of gold sodium thiosulfate in mice and guinea pigs. Biomaterials 23:4907-4914.

Y Ikarashi, Y Tsukamoto, T Tsuchiya, A Nakamura. 1993. Influence of irritants on lymph node cell proliferation and the detection of contact sensitivity to metal salts in the murine local lymph node assay. Contact Dermatitis 29:128-132.

Y Ikarashi, T Tsuchiya, A Nakamura. 1993. A sensitive mouse lymph node assay with two application phases for detection of contact allergens. Archives of Toxicology 67:629-636.

---. 1993. Evaluation of contact sensitivity of rubber chemicals using the murine local lymph node assay. Contact Dermatitis 28:77-80.

Y Ikarashi, K Ohno, T Tsuchiya, A Nakamura. 1992. Differences of draining lymph node cell proliferation among mice, rats and guinea pigs following exposure to metal allergens. Toxicology 76:283-292.

Y Ikarashi, T Tsuchiya, A Nakamura. 1992. Detection of contact sensitivity of metal salts using the murine local lymph node assay. Toxicology Letters 62:53-61.

I Kimber, A Bentley, J Hilton. 1990. Contact sensitization of mice to nickel sulphate and potassium dichromate. Contact Dermatitis 23:325-330.

J Lalko, A Api. 2006. Investigation of the dermal sensitization potential of various essential oils in the local lymph node assay. Food and Chemical Toxicology 44:739-746.

C Mandervelt, F Clottens, M Demedts, B Nemery. 1997. Assessment of the sensitization potential of five metal salts in the murine local lymph node assay. Toxicology 120:65-73.

A Nakamura, Y Kanazawa, H Sato, T Tsuchiya, Y Ikarashi, W De Jong, et al. 2003. Evaluation of allergic potential of rubber products: comparison of sample preparation methods for the testing of polymeric medical devices. Journal of Toxicology - Cutaneous and Ocular Toxicology 22:169-185.

C Ryan, L Cruse, R Skinner, R Dearman, I Kimber, G Gerberick. 2002. Examination of a vehicle for use with water soluble materials in the murine local lymph node assay. Food and Chemical Toxicology 40:1719-1725.

D Sailstad, J Tepper, D Doerfler, M Qasim, M Selgrade. 1994. Evaluation of an azo and two anthraquinone dyes for allergic potential. Fundamental and Applied Toxicology 23:569-577.

M Skold, A Karlberg, M Matura, A Borje. 2006. The fragrance chemical beta-caryophyllene - air oxidation and skin sensitization. Food and Chemical Toxicology 44:538-545.

M Skold, A Borje, E Harambasic, A Karlberg. 2004. Contact allergens formed on air exposure of linalool. Identification and quantification of primary and secondary oxidation products and the effect on skin sensitization. Chemical Research in Toxicology 17:1697-1705.

S Tinkle, J Antonini, B Rich, J Roberts, R Salmen, K DePree, et al. 2003. Skin as a route of exposure and sensitization in chronic beryllium disease. Environmental Health Perspectives 111:1202-1208.

T Yamano, M Shimizu, T Noda. 2006. Allergenicity and cross-reactivity of naphthenic acid and its metallic salts in experimental animals. Contact Dermatitis 54:25-28.