Human Cell Line Activation Test

Topic: Skin sensitisation

Test Method Number:

TM2008-05 (EU)

Short Name of TM:

hCLAT

Year received:

2008

Responsible Organisation:

EURL ECVAM - European Union

Protocol(s)/SOP(s):

158

General Comments:

Test Guideline has been adopted as TG442E in July 2016

Method Description

The h-CLAT supports the discrimination between sensitizing and non-sensitizing chemicals

The h-CLAT quantifies in vitro changes in the expression of the CD86 and CD54 membrane phenotypic markers in a human monocytic leukemia cell line (THP-1 cells).

THP-1 cells are monocyte-derived cells that have shown to produce DC-like responses following exposure to skin sensitising chemicals (e.g. upregulation of surface markes (e.g. CD86 and CD54) and cytokine production (e.g. TNF-α).

The CD86 (a co-stimulatory molecule) and the CD54 (an adhesion molecule) are upregulated in activated Dendritic Cells (DC) and play a critical role in DC presentation of antigens to T cells (T-cell priming).

By studying the potential of test chemicals to up-regulate markers of DC activation, the h-CLAT generates information addressing key event 3 (dendritic cell activation) of the skin sensitization Adverse Outcome Pathway.

Track Approval Status

Submission
Validation
Peer-review
Recommendation
Regulatory acceptance/Standards

Submission

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Date	Stage	Comment
Jan 1970	Assessment of full submission
Nov 2008	Receipt of presubmission	Pre-submission (TPF) received
Dec 2008	Receipt of full submission	Full submission received
Feb 2009	Assessment finalised	Submitter informed that the method qualifies for entering validation

Validation

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Date	Stage	Comment
May 2009	Planning	Invitation to enter phase II prevalidation of EURL ECVAM validation process together with DPRA and U-SENS
Nov 2009	Ongoing	Starting of EURL ECVAM coordinated validation study on the h-CLAT. The study was designed to generate information according to the modular approach to validation with the primary objective of fully assessing the reliability of the h-CLAT (i.e. its transferability and within and between laboratory reproducibility).
Jul 2013	Finalised	Validation step completed and validation study report prepared

Peer-review

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Date	Stage	Comment	Documents/Links
Sep 2009	Planning	First Validation Management Group (VMG) meeting
Feb 2013	Ongoing	First ESAC Working Group meeting to discuss initial findings, and resolve contentious points.	TM2008-05 hCLAT ESAC WG
Nov 2013	Ongoing	First version of this ESAC WG report.
Mar 2014	Ongoing	Endorsement of the WG peer review consensus report and the ESAC opinion at ESAC plenary meeting
May 2014	Finalised	Release of the WG Consensus Report. ESAC opinion included as annex in the EURL ECVAM recomendation (please see Recommendation step)	hCLAT Consensus Report

Recommendation

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Date	Stage	Comment	Documents/Links
Feb 2015	Published	EURL ECVAM Recommendation on the human Cell Line Activation Test (h-CLAT) for skin sensitisation testing. Annex I - ESAC Opinion	EURL ECVAM Recommendation

Regulatory acceptance/Standards

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Date	Stage	Comment	Documents/Links
Apr 2013	Drafting of new regulatory standard/guideline	Project approved and included in the OECD Test Guidelines Programme.
Jul 2016	Adopted/published	OECD TG 442E.	Test Guideline 442E