Toxicological Research

Korean Society of Toxicology & Korea Environmental Mutagen Society (한국독성학회)
권호 표지

A Non-radioisotopic Endpoint Using Bromodeoxyuridine ELISA Method for Murine Local Lymph Node Assay

BrdU ELISA를 이용한 국소 림프절 시험법의 비방사선법 연구

  • 이종권 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 박재현 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 박승희 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 김형수 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 정승태 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 엄준호 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 윤소미 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 장은정 (식품의약품안전청 국립독성연구원 특수독성부) ;
  • 최광식 (식품의약품안전청 국립독성연구원 특수독성부)
  • Published : 2003.06.01
Download PDF
⟨ Previous Next ⟩

Abstract

Allergic contact dermatitis may be caused by a wide variety of chemicals. A murine local lymph node assay (LLNA) has been developed as an alternative to guinea pig models for assessing the contact sensitization potential of chemical. However, there is a need to develop a nonradioisotopic endpoint for the LLNA, because of the radioisotopic method's requiring the use of special facilities. In this study, we investigated the development of a nonradioisotopic endpoint for LLNA using ELISA (enzyme-linked immunosorbent assay). Female Balb/c mice were treated by the topical application on the dorsum of both ears with four different strong sensitizers, 2,4-dinitrochlorobenzene (DNCB), oxazolone (OXZ), toluene diisocyanate (TDI), and trimellitic anhydride (TMA), and a strong irritant, sodium lauryl sulfate (SLS), once daily for three consecutive days. The proliferation of cells in the auricular Iymph node was analyzed by means of the labelling index (Ll) of bromodeoxyuridine (BrdU) incorporation into cells. The weights of the Iymph nodes in the mice treated with allergens, DNCB, OXZ, TDl and TMA were increased compared to the vehicle control. The stimulation index (Sl) of mice treated with DNCB, OXZ, TDl, and TMA was over three-fold increase compared to the vehicle control. However, the S1 of mice exposed to SLS was not significantly increased compared to the vehicle control, while the lymph node weight of SLS was significantly increased. These results suggest that the LLNA modified endpoint using ELISA based on BrdU incorporation could provide a useful method of screening for irritants and allergens.

Keywords

References

  1. Toxicology v.117 Local lymph node activation and lgE responses in Brown Norway and Wistar rats after dermal application of sensitizing and nonsensitizing chemicals Arts,J.H.E.;Droge,S.C.M.;Spanhaak,S.;Bloksma,N.;Peninks,A.H.;Kuper,C.F. https://doi.org/10.1016/S0300-483X(96)03576-7
  2. Fd. Chem.Toxicol. v.30 Comparison of the local lymph node assay with the guinea-pig maximization test for the detection of a range of contact allergens Basketter,D.A.;Scholes,E.W.
  3. Contact Dermatitis v.42 Use of local lymph node assay for the estimation of relative contact allergenic protency Basketter,D.A.;Blaikie,L.;Dearman,R.J.;Kimber,I.;Ryan,C.A.;Gerberick,G.F.;Harvey,P.;Evans,P.;White,I.R.;Rycroft,R.J.G. https://doi.org/10.1034/j.1600-0536.2000.042006344.x
  4. Fd. Chem. Toxicol. v.40 Local lymph node assay - validation, conduct and use in practice Basketter,D.A.;Evans,P.;Fielder,R.J.;Gerberick,G.F.;Kimber,I. https://doi.org/10.1016/S0278-6915(01)00130-2
  5. Fd. Chem. Toxicol. v.36 Strategies for identifying false positive responses in predictive skin sensitization tests Basketter,D.A.;Gerberick,G.F.;Kimber,I. https://doi.org/10.1016/S0278-6915(97)00158-0
  6. Food Chem. Toxicol. v.34 The local lymph node assay: A viable alternative to currently accepted skin sensitization tests Basketter,D.A.;Gerberick,G.F.;Kimber,I.;Loveless,S.E. https://doi.org/10.1016/S0278-6915(96)00059-2
  7. J. Appl. Toxicol. v.15 Evaluation of lymphocyte proliferation by immunohistochemistry in the local lymph node assay Bossiquet-Leroux,C.;Durand Cabagna,G.;Herlin,H.;Helder,D. https://doi.org/10.1002/jat.2550150608
  8. Food Chem. Toxicol. v.29 Skin sensitization- a critical review of predictive test methods in animals and man Botham,P.A.;Basketter,D.A.;Maurer,T.;Mueller,D.;Potokar,M.;Bontinck,W. https://doi.org/10.1016/0278-6915(91)90025-3
  9. Arch. Dermatol. v.91 Delayed contact hypersensitivity in guinea pig Buehler,E.V. https://doi.org/10.1001/archderm.1965.01600080079017
  10. Methods v.19 Cytokine fingerprinting: Characterization of chemical allergens Dearman,R.J.;Kimber,I. https://doi.org/10.1006/meth.1999.0827
  11. J. Applied Toxicol. v.17 Classification of chemical allegens according to cytokine secretion profiles of murine lymph node cells Dearman,R.J.;Smith,S.;Basketter,D.A.;Kimber,I.
  12. Cosmetics legislation v.1 Council Directive 76/778/EEC in CosmetLex EC(European Communities)
  13. Statistics for toxicologists in Principles and Methods of Toxicology Gad,S.C.;Weil,G.S.;Hayes,A.W.(ed.)
  14. Toxicol. Appl. Pharmacol v.84 Developmemt and validation of an alternative dermal sensitization test: The mouse ear swelling test (MEST) Gad,S.C.;Dunn,B.J.;Dobbs,D.W.;Christopher,R.;Walsh,R.D. https://doi.org/10.1016/0041-008X(86)90419-9
  15. Methods v.19 Local lymph node assay: Differentiating allergic and irritant responses using flowcytometry Gerberick,G.F.;Cruse,L.W.;Ryan,C.A. https://doi.org/10.1006/meth.1999.0826
  16. Toxicol. Sci. v.67 Use of a B cell marker (B220) to discrimate between allergens and irritants in the local lymph node assay Gerberick,G.F.;Cruse,L.W.;Ryan,C.A.;Hulette,B.C.;Chaney,J.G.;Skinner,R.A.;Deaman,R.J.;Kimber,I.
  17. Food Chem. Toxicol. v.37 Development of a non-radioactive endpoint in a modified local lymph node assay Haryia,T.;Hatao,M.;Ichikawa,H. https://doi.org/10.1016/S0278-6915(98)00102-1
  18. Food Chem. Toxicol. v.30 The murine local lymph node a commentary an collborative studies and new directions Kimber,I.;Basketter,D.A. https://doi.org/10.1016/0278-6915(92)90153-C
  19. Toxicol. Sci. v.59 Skin sensitization testing in potency and risk assessment Kimber,I.;Basketter,D.A.;Bethold,K.;Butler,M.;Garrigue,J.L.;Lea,L.;Newsome,C.;Roggeband,R.;Steiling,W.;Stropp,G.Waterman,S.;Wiemann,C. https://doi.org/10.1093/toxsci/59.2.198
  20. Toxicology v.93 The local lymph node assay: developments and applications Kimber,I.;Dearman,R.J.;Schole,E.W.Basketter,D.A.. https://doi.org/10.1016/0300-483X(94)90193-7
  21. Toxicology v.103 An international evaluation of the murine local lymph node assay and comparison of modified procedures Kimber,I.:Hilton,J.;Dearman,R.J.;Gerberick,G.F.;Ryan,C.A.;Baskette,D.A.;Edward,W.S.;Robert,V.H. https://doi.org/10.1016/0300-483X(95)03114-U
  22. Toxicol. Lett. v.106 lmmune responses to contact allergens novel approaches to hazard evaluation Kimber,I.;Pichowski,J.S.;Basketter,D.A.;Dearman,R.J. https://doi.org/10.1016/S0378-4274(99)00069-7
  23. J. Toxicol. Pub. Health v.18 Evaluation of local lymph node assay as an alternative method for skin sensitization potential in Balb/c mice Lee,J.K.;Hwang,I.C.;Park,J.H.;Kim,H.S.;Chung,S.T.;Eom,J.H.;Oh,H.Y.
  24. Environ. Toxicol. Pharmacol. Evaluation of cell proliferation in ear and lymph node using BrdU immunohistochemistry for mouse ear swelling test Lee,J.K.;Park,J.H.;Kim,H.S.;Chung,S.T.;Eom,J.H.;Nam,K.T.;Oh,H.Y.
  25. J. Pharmacol. Toxicol. Methods v.48 A nonradioisotopic endpoint for measurement of lymph node cell proliferation in a murine allergic contact dermatitis model, using bromodexoyuridine immunohistochemistry Lee,J.K.;Park,J.H.;Park,S.H.;Kim,H.S.;Oh,H.Y. https://doi.org/10.1016/S1056-8719(03)00021-2
  26. Toxicology v.108 Further evaluation of the local lymph node assay in the final phase of an international collaborative trial Loveless,S.E.;Gregory,S.L.;Gerberick,G.F.;Ryan,C.A. https://doi.org/10.1016/0300-483X(95)03279-O
  27. J. Invest. Dermatol. v.52 The identification of contact allergen by animal assay. The guinea pig maximization test Magnusson,B.;Kligman,A.M. https://doi.org/10.1038/jid.1969.42
  28. Toxicol. Appl. Pharmacol. v.171 The determination of draining lymph node cell cytokine mRNA levels in BALB/c mice following dermal sodium lauryl sulfate,dinitrofluorobenzene and toluene disocyanate exposure Manetz,T.S.;Pettit,D.A.;Meade,B.J. https://doi.org/10.1006/taap.2000.9123
  29. Fund. Appl. Toxicol. v.34 Phenotypic analysis of lymphocyte subpopulations in lymph nodes draining the ear following exposure to contact allergens and irritants Sikorski,E.E.;Gerberick,G.F.;Ryan,C.A.;Miller,C.M.;Ridder,G.M. https://doi.org/10.1006/faat.1996.0172
  30. J. Toxicol. Sci. v.27 Local lymph node assay with non-radioisotopic alternative endpoints Suda,A.;Yamashita,M.;Tabei,M.;Taguchi,K.;Vohr,H.;Tsusui,N.;Suzuki,K.;Kikuchi,K.;Sakaguchi,K.;Mochizuki,K.;Nakamura,N. https://doi.org/10.2131/jts.27.205
  31. Toxicol. Letters v.119 Development of non-radio isotopic endpoint of murine local lymph node assay based on 5-broo-2'-deoxyuridine (BrdU) incorporation Takeyoshi,M.;Yamaski,K.;Yakabe.Y.;Takatuski,M.;Kimber,I. https://doi.org/10.1016/S0378-4274(00)00315-5
  32. Toxicol. Appl. Pharmacol. v.162 Assessment of preferential T-Helper 1 or T-Helper 2 induction by low molecular weight compounds using the local lymph node assay in conjunction with RT-PCR and ELISA for interferon-γ and Interleukin-4 Vandebriel,R.H.;De Jong,W.H.;Spiekstra,S.W.;Mariskam,V.D.;Angelique,F.;Gassen,J. https://doi.org/10.1006/taap.1999.8841