A modification of the local lymph node assay for contact allergenicity screening: measurement of interleukin-2 as an alternative to radioisotope-dependent proliferation assay.
The local lymph node assay is an effective prediction method for contact allergenicity, but employs radioisotopes. We investigated whether measurement of interleukin-2 (IL-2) production by lymph node cells could be used instead to predict contact allergenicity of chemicals. Test chemicals were applied for three consecutive days to both ears of Balb/c mice and the auricular lymph nodes were obtained on either the fourth or fifth day after the first application. Both IL-2 concentration in supernatant of the suspension and proliferative activity of lymph node cells were determined after 24-, 48-, 72-h cell culture in RPMI-1640 medium by ELISA and by measuring [3H]methylthymidine incorporation, respectively. These two methods detected allergenicity similarly except in the case of TNCB and oxazolone, which showed excessive proliferation-inducing capacity as compared to IL-2 release-increasing effect. Flow cytometry showed that these two chemicals also increased the percentage of Iad-positive cells in the lymph nodes, suggesting that these chemicals might induce not only cellular immunity but also humoral immunity. We conclude that interleukin-2 assay is a convenient and dependable method for screening strong contact allergens without using radioisotopes.
Hatao
M
M
Shiseido Safety and Analytical Research Center, Yokohama, Japan.
Hariya
T
T
Katsumura
Y
Y
Kato
S
S
D000485:Allergens / Q000276:immunology*; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002455:Cell Division / Q000276:immunology; D017449:Dermatitis, Allergic Contact / Q000175:diagnosis*; D005260:Female; D005434:Flow Cytometry / Q000379:methods; D007376:Interleukin-2 / Q000096:biosynthesis*; D008198:Lymph Nodes / Q000166:cytology / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D010641:Phenotype; D013936:Thymidine / Q000031:analogs & derivatives; D014316:Tritium
D000485:Allergens; D015415:Biomarkers; D007376:Interleukin-2; D014316:Tritium; C012439:methylthymidine; D013936:Thymidine