PMID- 18462956
OWN - NLM
STAT- MEDLINE
DCOM- 20081028
LR  - 20131121
IS  - 1873-488X (Electronic)
IS  - 1056-8719 (Linking)
VI  - 58
IP  - 1
DP  - 2008 Jul-Aug
TI  - Characterization and evaluation of a modified local lymph node assay using ATP 
      content as a non-radio isotopic endpoint.
PG  - 1-10
LID - 10.1016/j.vascn.2008.03.003 [doi]
AB  - INTRODUCTION: The murine local lymph node assay (LLNA) is an accepted and widely 
      used method for assessing the skin-sensitizing potential of chemicals. Here, we 
      describe a non-radio isotopic modified LLNA in which adenosine triphosphate (ATP) 
      content is used as an endpoint instead of radioisotope (RI); the method is termed 
      LLNA modified by Daicel based on ATP content (LLNA-DA). METHODS: Groups of female 
      CBA/JNCrlj mice were treated topically on the dorsum of both ears with test 
      chemicals or a vehicle control on days 1, 2, and 3; an additional fourth 
      application was conducted on day 7. Pretreatment with 1% sodium lauryl sulfate 
      solution was performed 1 h before each application. On day 8, the amount of ATP 
      in the draining auricular lymph nodes was measured as an alternative endpoint by 
      the luciferin-luciferase assay in terms of bioluminescence (relative light units, 
      RLU). A stimulation index (SI) relative to the concurrent vehicle control was 
      derived based on the RLU value, and an SI of 3 was set as the cut-off value. 
      RESULTS: Using the LLNA-DA method, 31 chemicals were tested and the results were 
      compared with those of other test methods. The accuracy of LLNA-DA vs LLNA, 
      guinea pig tests, and human tests was 93% (28/30), 80% (20/25), and 79% (15/19), 
      respectively. The estimated concentration (EC) 3 value was calculated and 
      compared with that of the original LLNA. It was found that the EC3 values 
      obtained by LLNA-DA were almost equal to those obtained by the original LLNA. 
      DISCUSSION: The SI value based on ATP content is similar to that of the original 
      LLNA as a result of the modifications in the chemical treatment procedure, which 
      contribute to improving the SI value. It is concluded that LLNA-DA is a promising 
      non-RI alternative method for evaluating the skin-sensitizing potential of 
      chemicals.
FAU - Idehara, Kenji
AU  - Idehara K
AD  - Analysis Service Center, Daicel Chemical Industries, Ltd., 1239, Shinzaike, 
      Aboshi-ku, Himeji, Hyogo 671-1283, Japan. kn_idehara@daicel.co.jp
FAU - Yamagishi, Gaku
AU  - Yamagishi G
FAU - Yamashita, Kunihiko
AU  - Yamashita K
FAU - Ito, Michio
AU  - Ito M
LA  - eng
PT  - Journal Article
DEP - 20080326
PL  - United States
TA  - J Pharmacol Toxicol Methods
JT  - Journal of pharmacological and toxicological methods
JID - 9206091
RN  - 0 (Irritants)
RN  - 8L70Q75FXE (Adenosine Triphosphate)
SB  - IM
MH  - Adenosine Triphosphate/*metabolism
MH  - Animals
MH  - Dermatitis, Allergic Contact/diagnosis/*etiology
MH  - Disease Models, Animal
MH  - Endpoint Determination/methods
MH  - Female
MH  - Guinea Pigs
MH  - Humans
MH  - Irritants/*toxicity
MH  - *Local Lymph Node Assay
MH  - Mice
MH  - Mice, Inbred CBA
MH  - Reproducibility of Results
MH  - Species Specificity
MH  - Toxicity Tests
EDAT- 2008/05/09 09:00
MHDA- 2008/10/29 09:00
CRDT- 2008/05/09 09:00
PHST- 2007/08/18 00:00 [received]
PHST- 2008/03/18 00:00 [accepted]
PHST- 2008/05/09 09:00 [pubmed]
PHST- 2008/10/29 09:00 [medline]
PHST- 2008/05/09 09:00 [entrez]
AID - S1056-8719(08)00025-7 [pii]
AID - 10.1016/j.vascn.2008.03.003 [doi]
PST - ppublish
SO  - J Pharmacol Toxicol Methods. 2008 Jul-Aug;58(1):1-10. doi: 
      10.1016/j.vascn.2008.03.003. Epub 2008 Mar 26.