PMID- 25600862 OWN - NLM STAT- MEDLINE DCOM- 20151209 LR - 20150316 IS - 1873-488X (Electronic) IS - 1056-8719 (Linking) VI - 72 DP - 2015 Mar-Apr TI - Application of BALB/c mouse in the local lymph node assay:BrdU-ELISA for the prediction of the skin sensitizing potential of chemicals. PG - 53-8 LID - S1056-8719(15)00002-7 [pii] LID - 10.1016/j.vascn.2015.01.001 [doi] AB - INTRODUCTION: Allergic contact dermatitis (ACD) is a skin disease characterized by eczema and itching. A considerable proportion of chemicals induce ACD in humans. More than 10,000 substances should be tested for skin sensitization potential under the Registration, Evaluation, Authorization and Restriction of Chemical substances (REACH) regulation. The Local Lymph Node Assay (LLNA) has been designated as the first-choice in vivo assay for sensitization testing by REACH. The LLNA:BrdU-ELISA is a validated non-radioactive modification to the LLNA. For both the LLNA and the LLNA:BrdU-ELISA, CBA/JN mouse is the preferred mouse strain recommended in the regulatory guidelines. However, the availability of CBA/JN mouse in China is only limited to a few animal suppliers, which makes the mouse difficult to obtain. BALB/c mouse, which is widely commercially available, is considered for alternative use but it can only be used in the assay after it has been evaluated by formal validation study. Thus, a validation study was conducted in our laboratory to determine if BALB/c mouse could also be used in the LLNA:BrdU-ELISA. METHODS: Forty-three test substances including 32 LLNA sensitizers and 11 LLNA non-sensitizers, their vehicles and each concentration used were the same as that used in the formal validation study for the LLNA:BrdU-ELISA using CBA/JN mouse. Female BALB/c mice of 8-10 weeks old were randomly allocated to groups (four mice per group). The test substance (25 mul) or the vehicle alone was applied to the dorsum of both ears daily for 3 consecutive days. A single intraperitoneal injection of 0.5 ml of BrdU (10mg/ml) solution was given on day 5. On day 6, a pair of auricular lymph nodes from each mouse was excised, weighed and stored at -20 degrees C until BrdU-ELISA was conducted. RESULTS: This validation study for the LLNA:BrdU-ELISA using BALB/c mouse correctly identified 30 of 31 sensitizers and 8 of 11 non-sensitizers. The accuracy, sensitivity, specificity, false positive rate, false negative rate, positive predictivity values and negative predictivity values in this study, which could indicate the performance of the LLNA:BrdU-ELISA using BALB/c mouse, were not different statistically from that of the validation study for the LLNA:BrdU-ELISA using CBA/JN mouse. DISCUSSION: This validation study indicates that BALB/c mouse could be used alternatively in the LLNA:BrdU-ELISA for the prediction of the skin sensitizing potential of chemicals. CI - Copyright (c) 2015 Elsevier Inc. All rights reserved. FAU - Hou, Fenxia AU - Hou F AD - Department of Toxicology, National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing, China. Electronic address: houfenxia66@163.com. FAU - Xing, Caihong AU - Xing C AD - Department of Toxicology, National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing, China. FAU - Li, Bin AU - Li B AD - Department of Toxicology, National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing, China. FAU - Cheng, Juan AU - Cheng J AD - Department of Toxicology, National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing, China. FAU - Chen, Wei AU - Chen W AD - Department of Toxicology, National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing, China. FAU - Zhang, Man AU - Zhang M AD - Department of Toxicology, National Institute of Occupational Health and Poison Control, Chinese Center for Disease Control and Prevention, Beijing, China. LA - eng PT - Journal Article PT - Research Support, Non-U.S. Gov't DEP - 20150116 PL - United States TA - J Pharmacol Toxicol Methods JT - Journal of pharmacological and toxicological methods JID - 9206091 RN - 0 (Allergens) RN - G34N38R2N1 (Bromodeoxyuridine) SB - IM MH - Allergens/*toxicity MH - Animals MH - Bromodeoxyuridine/*metabolism MH - *Dermatitis, Allergic Contact MH - Enzyme-Linked Immunosorbent Assay MH - Female MH - *Local Lymph Node Assay MH - Lymph Nodes/cytology MH - Mice MH - Mice, Inbred BALB C MH - Mice, Inbred CBA MH - Predictive Value of Tests MH - Sensitivity and Specificity OTO - NOTNLM OT - 2,4-dinitrochlorobenzene (PubChem CID: 6) OT - 5-bromo-2-deoxyuridine (PubChem CID: 6035) OT - 5-chloro-2-methyl-4-isothiazolin-3-one (PubChem CID: 33344) OT - BALB/c mouse OT - BrdU-ELISA OT - diphenylcyclopropenone (PubChem CID: 65057) OT - eugenol (PubChem CID: 3314) OT - hexyl cinnamic aldehyde (PubChem CID: 1715135) OT - isopropanol (PubChem CID: 3776) OT - local lymph node assay OT - methods OT - methyl salicylate (PubChem CID: 4133) OT - p-benzoquinone (PubChem CID: 4650) OT - salicylic acid (PubChem CID: 338) OT - skin sensitization OT - validation EDAT- 2015/01/21 06:00 MHDA- 2015/12/15 06:00 CRDT- 2015/01/21 06:00 PHST- 2014/08/13 00:00 [received] PHST- 2014/12/05 00:00 [revised] PHST- 2015/01/05 00:00 [accepted] PHST- 2015/01/21 06:00 [entrez] PHST- 2015/01/21 06:00 [pubmed] PHST- 2015/12/15 06:00 [medline] AID - S1056-8719(15)00002-7 [pii] AID - 10.1016/j.vascn.2015.01.001 [doi] PST - ppublish SO - J Pharmacol Toxicol Methods. 2015 Mar-Apr;72:53-8. doi: 10.1016/j.vascn.2015.01.001. Epub 2015 Jan 16.