OECD 442E: h-CLAT
SERVICE INFORMATION
The OECD 442E human cell activation test (h-CLAT) investigates key event 3 of the skin sensitisation adverse outcome pathway – dendritic cell activation. Utilising the monocytic THP-1 cell line, the ability of a test substance to increase expression of CD54 & CD86 cell surface markers is assessed by flow cytometry. Upregulation of these cell surface markers is associated with dendritic cell activation and leads to the stimulation of an immune response required for sensitisation of the skin to the test substance.
h-CLAT protocols have been validated and laboratory proficiency under OECD 442E established at Gentronix, with services available in both GLP and non-GLP formats.
OECD guideline | OECD 442E |
Adverse Outcome Pathway Key Event | Number 3 – activation of dendritic cells |
What it measures | Increase in expression of cell surface markers (CD86 and CD54) measured by flow cytometry |
What is the relevance of that? | Expression of CD54 & CD86 is linked to the activation of dendritic cells, which is a precursor event for the stimulation of T-cell proliferation, leading to skin allergenicity |
Test System | human monocytic leukaemia cell line THP-1 |
Test Item Requirements | 1 g |
h-CLAT OECD 442E Proficiency Data
Gentronix values consistent with OECD reference range | Gentronix study calls consistent with OECD reference | |||||||
---|---|---|---|---|---|---|---|---|
Test chemical | In vivo classification | CV75 | CD86 | CD54 | CD86 Call | CD54 Call | h-CLAT classification | Classification correct |
2,4-Dinitrochlorobenzene | Sensitiser (extreme) | Positive | Positive | Sensitiser | ||||
4-Phenylenediamine | Sensitiser (strong) | Positive | Negative | Sensitiser | ||||
Nickel sulfate | Sensitiser (moderate) | Positive | Positive | Sensitiser | ||||
2-Mercaptbenzothiazole | Sensitiser (moderate) | Negative | Positive | Sensitiser | ||||
R(+)-Limonene | Sensitiser (weak) | Negative | Positive | Sensitiser | ||||
Imidazolidinyl urea | Sensitiser (weak) | Positive | Positive | Sensitiser | ||||
Isopropanol | Non-sensitiser | Negative | Negative | Non-sensitiser | ||||
Glycerol | Non-sensitiser | Negative | Negative | Non-sensitiser | ||||
Lactic acid | Non-sensitiser | Negative | Negative | Non-sensitiser | ||||
4-Aminobenzoic acid | Non-sensitiser | Negative | Negative | Non-sensitiser |