Performance standard-based validation study for local lymph node assay: 5-bromo-2-deoxyuridine-flow cytometry method.
Local lymph node assay: 5-bromo-2-deoxyuridine-flow cytometry method (LLNA: BrdU-FCM) is a modified non-radioisotopic technique with the additional advantages of accommodating multiple endpoints with the introduction of FCM, and refinement and reduction of animal use by using a sophisticated prescreening scheme. Reliability and accuracy of the LLNA: BrdU-FCM was determined according to OECD Test Guideline (TG) No. 429 (Skin Sensitization: Local Lymph Node Assay) performance standards (PS), with the participation of four laboratories. Transferability was demonstrated through successfully producing stimulation index (SI) values for 25% hexyl cinnamic aldehyde (HCA) consistently greater than 3, a predetermined threshold, by all participating laboratories. Within- and between-laboratory reproducibility was shown using HCA and 2,4-dinitrochlorobenzene, in which EC2.7 values (the estimated concentrations eliciting an SI of 2.7, the threshold for LLNA: BrdU-FCM) fell consistently within the acceptance ranges, 0.025-0.1% and 5-20%, respectively. Predictive capacity was tested using the final protocol version 1.3 for the 18 reference chemicals listed in OECD TG 429, of which results showed 84.6% sensitivity, 100% specificity, and 88.9% accuracy compared with the original LLNA. The data presented are considered to meet the performance criteria for the PS, and its predictive capacity was also sufficiently validated.
Korean Center for the Validation of Alternative Methods, National Institute of Food and Drug Safety Evaluation, MFDS, Chungbuk, Republic of Korea.
Ilyoung
I
Ahn
Korean Center for the Validation of Alternative Methods, National Institute of Food and Drug Safety Evaluation, MFDS, Chungbuk, Republic of Korea.
Tae-Sung
TS
Kim
National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Dept., Toxicological Screening & Testing Division, MFDS, Chungbuk, Republic of Korea.
Eun-Sun
ES
Jung
National Institute of Food and Drug Safety Evaluation, Toxicological Evaluation and Research Dept., Toxicological Screening & Testing Division, MFDS, Chungbuk, Republic of Korea.
Jung-Sun
JS
Yi
Medical Beauty Research Division, AmorePacific Corp., R&D Center, Yongin, Republic of Korea.
Won-Hee
WH
Jang
Medical Beauty Research Division, AmorePacific Corp., R&D Center, Yongin, Republic of Korea.
Kyoung-Mi
KM
Jung
Medical Beauty Research Division, AmorePacific Corp., R&D Center, Yongin, Republic of Korea.
Miyoung
M
Park
Biotoxtech Co., Ltd., Pharmacology Efficacy Team, Chungbuk, Republic of Korea.
Mi-Sook
MS
Jung
Biotoxtech Co., Ltd., Pharmacology Efficacy Team, Chungbuk, Republic of Korea.
Eun-Young
EY
Jeon
Catholic University of Daegu, College of Medical and Public Health Sciences, Dept. Occupational Health and GLP Center, Gyeongbuk, Republic of Korea.
Kyeong-Uk
KU
Yeo
Catholic University of Daegu, College of Medical and Public Health Sciences, Dept. Occupational Health and GLP Center, Gyeongbuk, Republic of Korea.
Ji-Hoon
JH
Jo
Catholic University of Daegu, College of Medical and Public Health Sciences, Dept. Occupational Health and GLP Center, Gyeongbuk, Republic of Korea.
Jung-Eun
JE
Park
Catholic University of Daegu, College of Medical and Public Health Sciences, Dept. Occupational Health and GLP Center, Gyeongbuk, Republic of Korea.
Chang-Yul
CY
Kim
Catholic University of Daegu, College of Medical and Public Health Sciences, Dept. Occupational Health and GLP Center, Gyeongbuk, Republic of Korea.
Yeong-Chul
YC
Park
Korean Center for the Validation of Alternative Methods, National Institute of Food and Drug Safety Evaluation, MFDS, Chungbuk, Republic of Korea.
Won-Keun
WK
Seong
Dongguk University, Ilsan Hospital, Gyeonggi-do, Republic of Korea.
Ai-Young
AY
Lee
Chung-Ang University, College of Pharmacy, Seoul, Republic of Korea.
Young Jin
YJ
Chun
Yeungnam University, College of Pharmacy, Gyeongbuk, Republic of Korea.
Tae Cheon
TC
Jeong
Chungbuk National University, College of Veterinary Medicine, Chungbuk, Republic of Korea.
Eui Bae
EB
Jeung
Ewha Womans University, College of Pharmacy, Seoul, Republic of Korea.
Kyung-Min
KM
Lim
Ewha Womans University, College of Pharmacy, Seoul, Republic of Korea.
SeungJin
S
Bae
Korean Center for the Validation of Alternative Methods, National Institute of Food and Drug Safety Evaluation, MFDS, Chungbuk, Republic of Korea.
Soojung
S
Sohn
Catholic University of Daegu, College of Medical and Public Health Sciences, Dept. Occupational Health and GLP Center, Gyeongbuk, Republic of Korea. Electronic address: yheo@cu.ac.kr.
Yong
Y
Heo
D000171:Acrolein / Q000031:analogs & derivatives* / Q000633:toxicity; D000818:Animals; D001973:Bromodeoxyuridine; D004137:Dinitrochlorobenzene / Q000633:toxicity*; D005260:Female; D005434:Flow Cytometry / Q000592:standards; D019983:Guideline Adherence; D017408:Guidelines as Topic; D006801:Humans; D059021:Laboratory Proficiency Testing; D023441:Local Lymph Node Assay; D008198:Lymph Nodes / Q000187:drug effects* / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D015588:Observer Variation; D011237:Predictive Value of Tests; D015203:Reproducibility of Results; D056910:Republic of Korea
D004137:Dinitrochlorobenzene; C080043:hexyl cinnamic aldehyde; D000171:Acrolein; D001973:Bromodeoxyuridine
Flow cytometry; LLNA: BrdU-FCM; Local lymph node assay; Performance standards; Skin sensitization; Validation study