PMID- 27456929
OWN - NLM
STAT- MEDLINE
DCOM- 20170515
LR  - 20170515
IS  - 1873-488X (Electronic)
IS  - 1056-8719 (Linking)
VI  - 82
DP  - 2016 Nov-Dec
TI  - Intra-laboratory study to determine the reproducibility of LLNA:BrdU-ELISA for 
      the prediction of the skin sensitizing potential of chemicals.
PG  - 26-30
LID - S1056-8719(16)30077-6 [pii]
LID - 10.1016/j.vascn.2016.07.003 [doi]
AB  - INTRODUCTION: The Local Lymph Node Assay (LLNA) has been designated as the 
      first-choice in vivo assay for identification the skin sensitization potential of 
      new chemicals. The LLNA:BrdU-ELISA is a validated non-radioactive modification to 
      the LLNA. An intra-laboratory reproducibility study for the LLNA:BrdU-ELISA was 
      conducted to demonstrate its adequate performance in our laboratory. METHODS: Ten 
      independent LLNA:BrdU-ELISAs with the preferred positive controls (PCs), i.e., 
      25% hexyl cinnamic aldehyde (HCA) and 25% eugenol, were conducted within a period 
      of less than one year. In addition, different concentrations of 
      2,4-dinitrochlorobenzene (DNCB, an extreme sensitizer) (0.01, 0.1 and 0.3%), HCA 
      (10, 25 and 50%) and eugenol (10, 25 and 50%), were tested to determine the EC1.6 
      values. Special Pathogen Free female CBA/J mice of 8-10weeks old were randomly 
      allocated to the groups, each group having 4 mice. 25mul of AOO (vehicle, acetone: 
      olive oil=4:1, v/v) or HCA, eugenol, DNCB at the needed concentration was applied 
      to the dorsum of each ear of the mice, daily for 3 consecutive days. A single 
      intraperitoneal injection of 0.5ml of BrdU solution (10mg/ml) was given on day 5. 
      On day 6, a pair of auricular lymph nodes from each mouse was excised, and BrdU 
      ELISA analysis was conducted. RESULTS: The result for each group is expressed as 
      the mean Stimulation Index (SI). The mean of the 10 mean SIs for 25% HCA 
      (2.58+/-0.95) and 25% eugenol (3.51+/-1.25) was not significantly different to that 
      from the Interagency Coordinating Committee on the Validation of Alternative 
      Methods (ICCVAM) (i.e., the data on the formal validation study for the 
      LLNA:BrdU-ELISA by the ICCVAM) (3.03+/-2.00 for 25% HCA, 6.13+/-6.06 for 25% eugenol) 
      (P>0.05), with even smaller Coefficient of Variations (CV) (36.8% for 25% HCA, 
      35.6% for 25% eugenol) than that from the ICCVAM (66.0% for 25% HCA, 98.8% for 
      25% eugenol). In addition, the EC1.6 values for HCA, eugenol and DNCB (15.2, 12.5 
      and 0.25%, respectively) were consistent with that from the ICCVAM (12.92, 8.85 
      and 0.34%, respectively). DISCUSSION: The results indicate that the reliability 
      for our laboratory to conduct the LLNA:BrdU-ELISA is successfully determined.
CI  - Copyright (c) 2016 Elsevier Inc. All rights reserved.
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 - 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 - 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.
LA  - eng
PT  - Journal Article
DEP - 20160722
PL  - United States
TA  - J Pharmacol Toxicol Methods
JT  - Journal of pharmacological and toxicological methods
JID - 9206091
RN  - 0 (Allergens)
RN  - 101-86-0 (hexyl cinnamic aldehyde)
RN  - 3T8H1794QW (Eugenol)
RN  - 7864XYD3JJ (Acrolein)
RN  - G34N38R2N1 (Bromodeoxyuridine)
SB  - IM
MH  - Acrolein/analogs & derivatives/pharmacology
MH  - Allergens/*toxicity
MH  - Animals
MH  - Bromodeoxyuridine/metabolism
MH  - *Dermatitis, Allergic Contact/diagnosis
MH  - Enzyme-Linked Immunosorbent Assay
MH  - Eugenol/pharmacology
MH  - Female
MH  - Laboratory Proficiency Testing/*methods
MH  - *Local Lymph Node Assay
MH  - *Lymph Nodes/cytology/immunology
MH  - Mice, Inbred CBA
MH  - Predictive Value of Tests
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
OTO - NOTNLM
OT  - 2,4-Dinitrochlorobenzene (PubChem CID: 6)
OT  - 5-Bromo-2-deoxyuridine (PubChem CID: 6035)
OT  - Acetone (PubChem CID:180)
OT  - Eugenol
OT  - Eugenol (PubChem CID: 3314)
OT  - Hexyl cinnamic aldehyde
OT  - Hexyl cinnamic aldehyde (PubChem CID: 1,715,135)
OT  - Local Lymph Node Assay:BrdU-ELISA
OT  - Methods
OT  - Reliability
OT  - Reproducibility
OT  - Skin sensitization
EDAT- 2016/10/18 06:00
MHDA- 2017/05/16 06:00
CRDT- 2016/07/27 06:00
PHST- 2016/04/07 00:00 [received]
PHST- 2016/06/25 00:00 [revised]
PHST- 2016/07/21 00:00 [accepted]
PHST- 2016/10/18 06:00 [pubmed]
PHST- 2017/05/16 06:00 [medline]
PHST- 2016/07/27 06:00 [entrez]
AID - S1056-8719(16)30077-6 [pii]
AID - 10.1016/j.vascn.2016.07.003 [doi]
PST - ppublish
SO  - J Pharmacol Toxicol Methods. 2016 Nov-Dec;82:26-30. doi: 
      10.1016/j.vascn.2016.07.003. Epub 2016 Jul 22.