Assessment of the skin sensitization potential of topical medicaments using the local lymph node assay: an interlaboratory evaluation.
The murine local lymph node assay (LLNA) is a method for the predictive identification of chemicals that have a potential to cause skin sensitization. Activity is measured as a function of lymph node cell (LNC) proliferative responses stimulated by topical application of test chemicals. Those chemicals that induce a threefold or greater increase in LNC proliferation compared with concurrent vehicle controls are classified as skin sensitizers. In the present investigations we have evaluated further the reliability and accuracy of the LLNA. In the context of an international interlaboratory trial the sensitization potentials of six materials with a history of use in topical medicaments have been evaluated: benzoyl peroxide, hydroquinone, penicillin G, streptomycin sulfate, ethylenediamine dihydrochloride, and methyl salicylate. Each chemical was analyzed in the LLNA by all five laboratories. Either the standard LLNA protocol or minor modifications of it were used. Benzoyl peroxide and hydroquinone, both human contact allergens, elicited strong LLNA responses in each laboratory. Penicillin G, another material shown previously to cause allergic contact dermatitis in humans, was also positive in all laboratories. Streptomycin sulfate induced equivocal responses, in that this material provoked a positive LLNA response in only one of the five laboratories, and then only at the highest concentration tested. Ethylenediamine dihydrochloride dissolved in a 3:1 mixture of acetone with water, or in 4:1 acetone:olive oil (one laboratory), was uniformly negative. However, limited further testing with the free base of ethylene diamine yielded a positive LLNA response when applied in acetone:olive oil (AOO). Finally, methyl salicylate, a nonsensitizing skin irritant, was negative at all test concentrations in each laboratory. Collectively these data serve to confirm that the local lymph node assay is sufficiently robust to yield equivalent results when performed independently in separate laboratories and indicate also that the LLNA is of value in assessing the skin sensitization potential of topical medicaments.
Kimber
I
I
Zeneca Central Toxicology Laboratory, Macclesfield, Cheshire, UK. IAN.KIMBER@APVXCI.ZENECA.COM
Hilton
J
J
Dearman
R J
RJ
Gerberick
G F
GF
Ryan
C A
CA
Basketter
D A
DA
Lea
L
L
House
R V
RV
Ladics
G S
GS
Loveless
S E
SE
Hastings
K L
KL
D000287:Administration, Topical; D000818:Animals; D003627:Data Interpretation, Statistical; D003877:Dermatitis, Contact / Q000473:pathology*; D004342:Drug Hypersensitivity / Q000473:pathology*; D005260:Female; D008198:Lymph Nodes / Q000187:drug effects*; D051379:Mice; D008808:Mice, Inbred CBA; D011237:Predictive Value of Tests