Suitability of macrophage inflammatory protein-1beta production by THP-1 cells in differentiating skin sensitizers from irritant chemicals.
BACKGROUND
Worldwide restrictions in animal use for research have driven efforts to develop alternative methods.


OBJECTIVE
The study aimed to test the efficacy of the macrophage inflammatory protein-1beta (MIP-1beta) assay for testing chemicals' skin-sensitizing capacity.


METHODS
The assay was performed using 9 chemicals judged to be sensitizing and 7 non-sensitizing by the standard in vivo assays. THP-1 cells were cultured in the presence or absence of 4 doses, 0.01x, 0.1x, 0.5x, or 1x IC(50) (50% inhibitory concentration for THP-1 cell proliferation) of these chemicals for 24 hr, and the MIP-1beta level in the supernatants was determined. Skin sensitization by the test chemicals was determined by MIP-1beta production rates. The MIP-1beta production rate was expressed as the relative increase in MIP-1beta production in response to chemical treatment compared with vehicle treatment.


CONCLUSIONS
When the threshold MIP-1beta production rate used was 100% or 105% of dimethyl sulfoxide, all the sensitizing chemicals tested (dinitrochlorobenzene, hexyl cinnamic aldehyde, eugenol, hydroquinone, dinitrofluorobenzene, benzocaine, nickel, chromium, and 5-chloro-2-methyl-4-isothiazolin-3-one) were positive, and all the non-sensitizing chemicals (methyl salicylate, benzalkonium chloride, lactic acid, isopropanol, and salicylic acid), with the exception of sodium lauryl sulfate, were negative for MIP-1beta production. These results indicate that MIP-1beta could be a biomarker for classification of chemicals as sensitizers or non-sensitizers.
Lim
Yeon-Mi
YM
Catholic University of Deagu, 330 Kumrak 1 ri, Hayang eup, Kyongsan si, Kyongbuk 712-702, Korea.
Moon
Seong-Joon
SJ
An
Su-Sun
SS
Lee
Soo-Jin
SJ
Kim
Seo-Young
SY
Chang
Ih-Seop
IS
Park
Kui-Lea
KL
Kim
Hyoung-Ah
HA
Heo
Yong
Y
D000485:Allergens / Q000494:pharmacology* / Q000633:toxicity; D000826:Animal Testing Alternatives / Q000379:methods*; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D015415:Biomarkers / Q000378:metabolism; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D054407:Chemokine CCL4 / Q000096:biosynthesis*; D004797:Enzyme-Linked Immunosorbent Assay; D006168:Guinea Pigs; D006801:Humans; D020128:Inhibitory Concentration 50; D007509:Irritants / Q000494:pharmacology* / Q000633:toxicity; D009000:Monocytes / Q000187:drug effects* / Q000378:metabolism; D012867:Skin / Q000187:drug effects; D018709:Statistics, Nonparametric; D013778:Tetrazolium Salts; D013844:Thiazoles
D000485:Allergens; D015415:Biomarkers; D054407:Chemokine CCL4; D007509:Irritants; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue