title: Use of IL-8 release and p38 MAPK activation in THP-1 cells to identify allergens and to assess their potency in vitro.
abstract: The local lymph node assay (LLNA) has been developed to assess skin sensitization, and based on the EC3 value, it can also be used to evaluate allergen potency. Therefore, in the development of in vitro alternatives to the LLNA assay, one should not only consider the hazard identification but also the possibility to classify allergens relatively to their potency. We have recently described a selective release of interleukin-8 (IL-8) by chemical allergens in THP-1 cell line, and identified the activation of p38 mitogen-activated protein kinase (p38 MAPK) as a common pathway. Therefore, the purpose of this study was to expand the number of chemicals tested and to investigate whether IL-8 production and p38 MAPK activation can be used to classify allergens according to their potency. THP-1 cells were exposed to the contact allergens (p-benzoquinone, 2-aminophenol, isoeugenol, diethyl maleate, citral and imidazolidinyl urea), selected according to their potency in the LLNA, and to lactic acid and propylene glycol as non-sensitizers. p38 MAPK activation was evaluated 5-15 min after treatment by FACS analysis, while IL-8 release was assed by ELISA following 24h of incubation. p38 MAPK was activated by all contact allergens, including the pro-apten isoeugenol, whereas IL-8 release was significantly increased after stimulation with all allergens tested, except for isoeugenol. The failure of isoeugenol may be due to decrease in the stability of IL-8 mRNA. Irritants exposure, as expected, failed to induce both p38 MAPK activation and IL-8 release. A significant correlation between IL-8 release and the LLNA EC(3) was found (Pearson correlation r=0.743, p=0.0036, n=12). On the contrary, the activation of p38 MAPK showed no significant correlation between LLNA data and vigor of p38 MAPK activation. Overall, data presented confirm our previous observations and reveal IL-8 as potential tool not only to identify sensitizers, with the exception of pro-haptens, but also to classify them according to their potency, while p38 MAPK activation allows the identification of all sensitizers, including pro-haptens, but was not useful for potency classification.
authors.affiliation: Departamento Fisiologia, Facultat de Farmàcia, Universitat de Barcelona, Spain.
authors.forename: M
authors.initials: M
authors.lastname: Mitjans
authors.forename: V
authors.initials: V
authors.lastname: Galbiati
authors.forename: L
authors.initials: L
authors.lastname: Lucchi
authors.forename: B
authors.initials: B
authors.lastname: Viviani
authors.forename: M
authors.initials: M
authors.lastname: Marinovich
authors.forename: C L
authors.initials: CL
authors.lastname: Galli
authors.forename: E
authors.initials: E
authors.lastname: Corsini
mesh_terms: D000485:Allergens / Q000145:classification / Q000633:toxicity*; D000826:Animal Testing Alternatives; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines; D004306:Dose-Response Relationship, Immunologic; D004789:Enzyme Activation; D015870:Gene Expression; D006801:Humans; D016209:Interleukin-8 / Q000378:metabolism*; D009000:Monocytes / Q000187:drug effects* / Q000201:enzymology / Q000276:immunology; D012333:RNA, Messenger / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism*
chemical_list: D000485:Allergens; D016207:Cytokines; D016209:Interleukin-8; D012333:RNA, Messenger; D048051:p38 Mitogen-Activated Protein Kinases