Cytokine induction as a measure of cutaneous toxicity in primary and immortalized porcine keratinocytes exposed to jet fuels, and their relationship to normal human epidermal keratinocytes.
The purpose of this study was to identify biomarkers of toxicity in primary porcine keratinocytes (PKC) and an immortalized porcine keratinocyte cell line (MSK3877) exposed to jet fuels Jet A, JP-8, and JP-8+100. Cells were exposed to 0.1% jet fuels and assayed for interleukin-8 (IL-8) and tumor necrosis factor alpha (TNF-alpha) mRNA using the TaqMan real time quantitative reverse transcriptase PCR assay. IL-8 and TNF-alpha protein release was measured using an ELISA. PKC exposed to jet fuels caused a slight upregulation of TNF-alpha mRNA at early time points, but no significant differences in TNF-alpha protein production were detected. IL-8 mRNA was increased at 4 h following exposure, and IL-8 protein was increased at 8 h. In MSK 3877 cells, jet fuels were shown to increase the production and expression of TNF-alpha mRNA and protein at 30 min and 1 h following exposure, respectively. IL-8 mRNA was only slightly induced compared to control. IL-8 protein release was suppressed by jet fuel exposure. These results were compared with those of a previous study in our laboratory to evaluate the utility of using porcine cells in lieu of normal human epidermal keratinocytes (NHEK). Similarities exist between PKC and NHEK with respect to both TNF-alpha and IL-8 production. The expression profile of TNF-alpha in MSK3877 cells mimics that of NHEK. In contrast, the profile of IL-8 expression opposes that of PKC and NHEK. These results suggest that porcine keratinocytes are susceptible to jet fuel toxicity. However, the responses of immortalized cells may vary from those of PKC and NHEK necessitating cautious interpretation of such data.
Department of Clinical Sciences, Center for Cutaneous Toxicology and Residue Pharmacology, North Carolina State University, 4700 Hillsborough Street, Raleigh, NC 27606, USA.
D G
DG
Allen
J E
JE
Riviere
N A
NA
Monteiro-Riviere
D000704:Analysis of Variance; D000818:Animals; D015415:Biomarkers; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005648:Fuel Oils / Q000633:toxicity*; D006801:Humans; D016209:Interleukin-8 / Q000096:biosynthesis*; D015603:Keratinocytes / Q000187:drug effects* / Q000378:metabolism*; D012333:RNA, Messenger / Q000302:isolation & purification; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D015415:Biomarkers; D005648:Fuel Oils; D016209:Interleukin-8; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha