人细胞系激发试验(h-CLAT)评价化学物致敏性的研究
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- 英文论文题名：Skin sensitization evaluation of chemicals using Human Cell Line Activation Test(h-CLAT)
- 论文作者：陈田 ; 马珅俊 ; 章瑶 ; 廖峰 ; 杜军
- 英文论文作者：Tian Chen ; Shenjun Ma ; Yao Zhang ; Feng Liao ; Jun Du ; Amway(China) Research and Development Center
- 年：2016
- 作者机构：安利(中国)研发中心;
- 论文关键词：皮肤致敏 ; 替代方法 ; THP-1细胞 ; CD86 ; CD54
- 英文论文关键词：Skinsensitization ; Alternative method ; THP-1 cell ; CD86 ; CD54
- 会议召开时间：2016-06-15
- 会议录名称：第十一届中国化妆品学术研讨会论文集
- 英文会议录名称：The Proceedings of the 11th Seminar on Cosmetic China
- 语种：中文
- 分类号：TQ658
- 学会代码：ZGXE
- 会议名称：第十一届中国化妆品学术研讨会
- 会议地点：中国上海
- 主办单位：中国香料香精化妆品工业协会
- 学会名称：中国香料香精化妆品工业协会
- 页数：6
- 文件大小：422k
- 原文格式：O
- 会议级别：全国
摘要
采用体外培养的人急性单核细胞白血病细胞株THP-1进行人细胞系激发试验(h-CLAT),探讨该方法在化妆品原料皮肤致敏评价中的应用。选用8个已知皮肤致敏特性的参照物处理THP-1细胞,根据共刺激分子CD86和CD54的诱导表达情况,判定受试物是否具有潜在皮肤致敏性。结果显示,当以致敏物处理细胞后,THP-1细胞CD86和/或CD54表达明显上调,而非致敏物处理后,CD86和CD54的表达变化不明显。以细胞活力≥50%,RFI CD86≥150和/或RFI CD54≥200来判定是否为致敏物。结果提示,h-CLAT能准确区分致敏物和非致敏物。因此,h-CLAT能够较准确地预测化学物的皮肤致敏性,可用于化妆品原料皮肤致敏性的快速初筛。
Establishing human cell line activation test(h-CLAT) with THP-1 cell line,a human monocyte cell lymphoma,to evaluate the application of this in vitro method on cosmetic ingredient safety assessment.After exposure of 8 reference chemicals to THP-1 cell,skin sensitization potency of these chemicals was evaluated according to the inductive expression of co-stimulatory molecules,CD86 and CD54.A chemical will be considered as an allergen if relative fluorescence intensity(RFI) of CD86≥150 and/or CD54≥200with cell viability more than 50%.With these acceptance criteria,all the chemicals could be classified as allergen or non-allergen correctly.It can be concluded that h-CLAT,a valuable in vitro model for the identification of potential sensitizers,can be used in screening of cosmetic materials.
Establishing human cell line activation test(h-CLAT) with THP-1 cell line,a human monocyte cell lymphoma,to evaluate the application of this in vitro method on cosmetic ingredient safety assessment.After exposure of 8 reference chemicals to THP-1 cell,skin sensitization potency of these chemicals was evaluated according to the inductive expression of co-stimulatory molecules,CD86 and CD54.A chemical will be considered as an allergen if relative fluorescence intensity(RFI) of CD86≥150 and/or CD54≥200with cell viability more than 50%.With these acceptance criteria,all the chemicals could be classified as allergen or non-allergen correctly.It can be concluded that h-CLAT,a valuable in vitro model for the identification of potential sensitizers,can be used in screening of cosmetic materials.
引文
[1]Adler S,BasketterD,CretonS,and et al.Alternative(non-animal)methods for cosmetics testing:current status and future prospects-2010[J].Arch Toxicol,2011,85(5):367-485.
[2]Liebsch M,Grune B,Seiler A,and et al.Alternatives to animal testing:current status and future perspectives[J].Arch Toxicol.2011,85(8):841-858.
[3]仲少敏,陈璨,弓月等.人重复性损伤性斑贴试验评价化妆品安全性[J].中国美容医学,2012,21(9):1342-1345.
[4]Alves VM,Muratov E,Fourches D,et al.Predicting chemically-induced skin reactions.PartⅠ:QSAR models of skin sensitization and their application to identify potentially hazardous compounds.ToxicolApplPharmacol.2015,284(2):262-72.
[5]Gerberick GF,Vassallo JD,Bailey RE,et al.Development of a peptide reactivity assay for screening contact allergens.ToxicolSci,2004,81(2):332-43.
[6]Ashikaga T,Yoshida Y,Hirota M,et al.Development of an in vitro skin sensitization test using human cell lines:the human Cell Line Activation Test(h-CLAT).I.Optimization of the h-CLAT protocol.ToxicolIn Vitro.2006,20(5):767-73.
[7]Piroird C,Ovigne JM,Rousset F,et al.The Myeloid U937 Skin Sensitization Test(U-SENS)addresses the activation of dendritic cell event in the adverse outcome pathway for skin sensitization.ToxicolIn Vitro.2015,29(5):901-16.
[8]Hitzler M,Bergert A,Luch A,et al.Evaluation of selected biomarkers for the detection of chemical sensitization in human skin:a comparative study applying THP-1,MUTZ-3 and primary dendritic cells in culture.ToxicolIn Vitro.2013,27(6):1659-69.
[9]Cottrez F,Boitel E,Ourlin JC,et al.SENS-IS,a 3D reconstituted epidermis based model for quantifying chemical sensitization potency:Reproducibility and predictivity results from an inter-laboratory study.ToxicolIn Vitro.2016,32:248-260.
[10]dos Santos GG,Reinders J,Ouwehand K,et al.Progress on the development of human in vitro dendritic cell based assays for assessment of the sensitizing potential of a compound.ToxicolApplPharmacol.2009,236(3):372-82.
[11]Strickland J,Zang Q,Kleinstreuer N,et al.Integrated decision strategies for skin sensitization hazard.J ApplToxicol.2016.doi:10.1002/jat.3281.
[12]Sakaguchi H,Ryan C,Ovigne JM,et al.Predicting skin sensitization potential and inter-laboratory reproducibility of a human Cell Line Activation Test(h-CLAT)in the European Cosmetics Association(COLIPA)ring trials.ToxicolIn Vitro.2010,24(6):1810-20.
[2]Liebsch M,Grune B,Seiler A,and et al.Alternatives to animal testing:current status and future perspectives[J].Arch Toxicol.2011,85(8):841-858.
[3]仲少敏,陈璨,弓月等.人重复性损伤性斑贴试验评价化妆品安全性[J].中国美容医学,2012,21(9):1342-1345.
[4]Alves VM,Muratov E,Fourches D,et al.Predicting chemically-induced skin reactions.PartⅠ:QSAR models of skin sensitization and their application to identify potentially hazardous compounds.ToxicolApplPharmacol.2015,284(2):262-72.
[5]Gerberick GF,Vassallo JD,Bailey RE,et al.Development of a peptide reactivity assay for screening contact allergens.ToxicolSci,2004,81(2):332-43.
[6]Ashikaga T,Yoshida Y,Hirota M,et al.Development of an in vitro skin sensitization test using human cell lines:the human Cell Line Activation Test(h-CLAT).I.Optimization of the h-CLAT protocol.ToxicolIn Vitro.2006,20(5):767-73.
[7]Piroird C,Ovigne JM,Rousset F,et al.The Myeloid U937 Skin Sensitization Test(U-SENS)addresses the activation of dendritic cell event in the adverse outcome pathway for skin sensitization.ToxicolIn Vitro.2015,29(5):901-16.
[8]Hitzler M,Bergert A,Luch A,et al.Evaluation of selected biomarkers for the detection of chemical sensitization in human skin:a comparative study applying THP-1,MUTZ-3 and primary dendritic cells in culture.ToxicolIn Vitro.2013,27(6):1659-69.
[9]Cottrez F,Boitel E,Ourlin JC,et al.SENS-IS,a 3D reconstituted epidermis based model for quantifying chemical sensitization potency:Reproducibility and predictivity results from an inter-laboratory study.ToxicolIn Vitro.2016,32:248-260.
[10]dos Santos GG,Reinders J,Ouwehand K,et al.Progress on the development of human in vitro dendritic cell based assays for assessment of the sensitizing potential of a compound.ToxicolApplPharmacol.2009,236(3):372-82.
[11]Strickland J,Zang Q,Kleinstreuer N,et al.Integrated decision strategies for skin sensitization hazard.J ApplToxicol.2016.doi:10.1002/jat.3281.
[12]Sakaguchi H,Ryan C,Ovigne JM,et al.Predicting skin sensitization potential and inter-laboratory reproducibility of a human Cell Line Activation Test(h-CLAT)in the European Cosmetics Association(COLIPA)ring trials.ToxicolIn Vitro.2010,24(6):1810-20.