PMID- 21275877
OWN - NLM
STAT- MEDLINE
DCOM- 20110822
LR  - 20201209
IS  - 1547-6901 (Electronic)
IS  - 1547-691X (Linking)
VI  - 8
IP  - 2
DP  - 2011 Jun
TI  - Evaluation of auricular lymph node cell lymphocyte proliferation and cytokine 
      production as non-radioactive endpoints during murine contact allergy.
PG  - 131-9
LID - 10.3109/1547691X.2010.547996 [doi]
AB  - The murine local lymph node assay (LLNA) has been developed as a test method to 
      assess allergic contact dermatitis. In spite of the validity of the LLNA, 
      attention was drawn to the two disadvantages: use of radioactivity for in vivo 
      measurement of lymph node cell proliferation ([(3)H]-thymidine labeling) and the 
      possibility of false positive results caused by non-specific cell activation as a 
      result of inflammatory processes in the skin (irritation). We aimed to 
      investigate the following non-radioactive endpoints of LLNA: 
      5-bromo-2'-deoxyuridine (BrdU) incorporation ex vivo and in vivo, in vivo and ex 
      vivo cytokine production with or without phytohemagglutinin (PHA) stimulation. 
      Here, 8-12-week-old female BALB/c mice were treated topically with the strong 
      sensitizer 2,4-dinitrochlorobenzene (DNCB) in acetone:olive oil (AOO, 4:1 [v/v]) 
      at levels of 0.025, 0.05, 0.01, or 0.25% (w/v). Ear thickness was also measured 
      to determine the differentiation index (DI) indicating the proportion of 
      non-specific activation due to irritating properties of test compound. At the 
      concentration of 0.05%, stimulation index (SI) value was found to be 3 for DNCB 
      based on in vivo and ex vivo BrdU incorporation. The results of the in vivo and 
      ex vivo non-radioactive LLNA assays were compatible both with each other and with 
      previous radioactive LLNA data. Our results indicate that non-radioactive 
      endpoints may be used as an alternative to the [(3)H]-thymidine LLNA. The levels 
      of T(H)1 cytokines (IL-2 and IFNgamma) and T(H)2 cytokines (IL-4 and IL-5) in lymph 
      node cell cultures were significantly (P < 0.01) increased when DNCB was applied 
      at the concentrations of 0.05 and 0.1%, respectively. As the DI was > 1, the 
      applied concentrations of DNCB caused only allergic effect but not any irritant 
      effect. This study reports that the use of these non-radioactive endpoints can 
      assess allergic contact dermatitis caused by chemicals.
FAU - Ulker, Ozge Cemiloglu
AU  - Ulker OC
AD  - Faculty of Pharmacy, Department of Toxicology, Ankara University, Ankara, Turkey.
FAU - Atak, Aysegul
AU  - Atak A
FAU - Ates, Ilker
AU  - Ates I
FAU - Karakaya, Asuman
AU  - Karakaya A
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20110131
PL  - England
TA  - J Immunotoxicol
JT  - Journal of immunotoxicology
JID - 101201960
RN  - 0 (Cytokines)
RN  - 0 (Dinitrochlorobenzene)
RN  - 0 (Irritants)
SB  - IM
MH  - Animals
MH  - *Cell Proliferation
MH  - Cells, Cultured
MH  - Cytokines/*immunology
MH  - Dermatitis, Atopic/chemically induced/*immunology/pathology
MH  - Dinitrochlorobenzene/adverse effects/pharmacology
MH  - Female
MH  - Irritants/adverse effects/pharmacology
MH  - Lymph Nodes/*immunology/pathology
MH  - Mice
MH  - Mice, Inbred BALB C
MH  - Skin/immunology/pathology
MH  - Th1 Cells/*immunology/pathology
MH  - Th2 Cells/*immunology/pathology
EDAT- 2011/02/01 06:00
MHDA- 2011/08/23 06:00
CRDT- 2011/02/01 06:00
PHST- 2011/02/01 06:00 [entrez]
PHST- 2011/02/01 06:00 [pubmed]
PHST- 2011/08/23 06:00 [medline]
AID - 10.3109/1547691X.2010.547996 [doi]
PST - ppublish
SO  - J Immunotoxicol. 2011 Jun;8(2):131-9. doi: 10.3109/1547691X.2010.547996. Epub 
      2011 Jan 31.