PMID- 29695103
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20191120
IS  - 2073-4409 (Print)
IS  - 2073-4409 (Electronic)
IS  - 2073-4409 (Linking)
VI  - 7
IP  - 5
DP  - 2018 Apr 24
TI  - High-Throughput GLP-Capable Target Cell Visualization Assay for Measuring 
      Cell-Mediated Cytotoxicity.
LID - 10.3390/cells7050035 [doi]
LID - 35
AB  - One of the primary effector functions of immune cells is the killing of 
      virus-infected or malignant cells in the body. Natural killer (NK) and CD8 
      effector T cells are specialized for this function. The gold standard for 
      measuring such cell-mediated cytolysis has been the chromium release assay, in 
      which the leakage of the radioactive isotope from damaged target cells is being 
      detected. Flow cytometry-based single cell analysis of target cells has recently 
      been established as a non-radioactive alternative. Here we introduce a target 
      cell visualization assay (TVA) that applies similar target cell staining 
      approaches as used in flow cytometry but based on single cell computer image 
      analysis. Two versions of TVA are described here. In one, the decrease in numbers 
      of calcein-stained, i.e., viable, target cells is assessed. In the other, the 
      CFSE/PI TVA, the increase in numbers of dead target cells is established in 
      addition. TVA assays are shown to operate with the same sensitivity as standard 
      chromium release assays, and, leaving data audit trails in form of scanned (raw), 
      analyzed, and quality-controlled images, thus meeting requirements for measuring 
      cell-mediated cytolysis in a regulated environment.
FAU - Welter, Anna
AU  - Welter A
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. anna.welter@web.de.
AD  - Institute of Anatomy and Cell Biology, Friedrich-Alexander University 
      Erlangen-Nurnberg, 91054 Erlangen, Germany. anna.welter@web.de.
FAU - Sundararaman, Srividya
AU  - Sundararaman S
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. svidya.sraman@gmail.com.
FAU - Li, Ruliang
AU  - Li R
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. ruliang.li@immunospot.com.
FAU - Zhang, Ting
AU  - Zhang T
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. ting.zhang@immunospot.com.
FAU - Karulin, Alexey Y
AU  - Karulin AY
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. alexey.karulin@immunospot.com.
FAU - Lehmann, Alexander
AU  - Lehmann A
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. alexander.lehmann@immunospot.com.
FAU - Naeem, Villian
AU  - Naeem V
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. villian.naeem@immunospot.com.
FAU - Roen, Diana R
AU  - Roen DR
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. diana.roen@immunospot.com.
FAU - Kuerten, Stefanie
AU  - Kuerten S
AUID- ORCID: 0000-0001-6119-605X
AD  - Institute of Anatomy and Cell Biology, Friedrich-Alexander University 
      Erlangen-Nurnberg, 91054 Erlangen, Germany. stefanie.kuerten@fau.de.
FAU - Lehmann, Paul V
AU  - Lehmann PV
AD  - Research & Development Department, Cellular Technology Limited, Shaker Heights, 
      OH 44122, USA. paul.lehmann@immunospot.com.
LA  - eng
PT  - Journal Article
DEP - 20180424
PL  - Switzerland
TA  - Cells
JT  - Cells
JID - 101600052
PMC - PMC5981259
OTO - NOTNLM
OT  - ADCC
OT  - CD8 cells
OT  - CFR-Part 11 compliance
OT  - NK measurement
OT  - TVA
OT  - audit trails
OT  - cellular assay validation
OT  - immune monitoring
OT  - multi-color single cell imaging
COIS- P.V.L. is Founder, President and CEO of CTL, a company that specializes in immune 
      monitoring by ELISPOT and TVA. T.Z., V.N., R.L., A.Y.K., D.R.R., and A.L. are 
      employees of CTL, S.S. was a CTL employee while contributing to this study. A.W. 
      was a visiting scholar at CTL while she performed the experiments towards this 
      study. Based on the results reported here TVA has become a product of CTL offered 
      to the public.
EDAT- 2018/04/27 06:00
MHDA- 2018/04/27 06:01
PMCR- 2018/05/01
CRDT- 2018/04/27 06:00
PHST- 2018/03/27 00:00 [received]
PHST- 2018/04/19 00:00 [revised]
PHST- 2018/04/21 00:00 [accepted]
PHST- 2018/04/27 06:00 [entrez]
PHST- 2018/04/27 06:00 [pubmed]
PHST- 2018/04/27 06:01 [medline]
PHST- 2018/05/01 00:00 [pmc-release]
AID - cells7050035 [pii]
AID - cells-07-00035 [pii]
AID - 10.3390/cells7050035 [doi]
PST - epublish
SO  - Cells. 2018 Apr 24;7(5):35. doi: 10.3390/cells7050035.