PMID- 23662898
OWN - NLM
STAT- MEDLINE
DCOM- 20131115
LR  - 20211021
IS  - 1940-9818 (Electronic)
IS  - 0736-6205 (Print)
IS  - 0736-6205 (Linking)
VI  - 54
IP  - 5
DP  - 2013 May
TI  - Pairwise agonist scanning-flow cytometry (PAS-FC) measures inside-out signaling 
      and patient-specific response to combinatorial platelet agonists.
PG  - 271-7
LID - 10.2144/000114027 [doi]
AB  - Understanding the response of cells to multiple stimuli is vital for predicting 
      donor specific responses and better understanding the signaling pathways 
      involved. This is of particular importance in platelets because exposure of 
      phosphatidylserine (PS) occurs upon costimulation but not with a single agonist. 
      Here, we describe a multiplexed pairwise agonist scanning-flow cytometry (PAS-FC) 
      method of measuring platelet inside-out responses to all pairs of six platelet 
      agonists (convulxin, SFLLRN, AYPGKF, ADP, U46619, and PGE(2)) used at their 
      EC(50) concentrations. These agonists allowed exploration of platelet signaling 
      downstream of GPVI, PAR-1, PAR-4, P2Y(1), P2Y(12), TP, and IP receptors. The 
      three-color flow cytometry method simultaneously measured integrin alpha(IIb)beta(3) 
      activation with PAC-1 antibody, P-selectin exposure (via alpha granule release) with 
      anti-P-selectin, and PS exposure with annexin V. These responses were consistent 
      across a healthy male donor pool. In duplicate measurements with each donor, 4 of 
      the 10 donors had a sufficiently unique 45-parameter (15 pairs x 3 colors) 
      phenotype to self-cluster (P < 0.001). This method has the potential for 
      efficiently scanning for patient specific responses across a broad 
      agonist-receptor space.
FAU - Jaeger, Daniel T L
AU  - Jaeger DT
AD  - Institute for Medicine and Engineering, Department of Chemical and Biomolecular 
      Engineering, University of Pennsylvania, Philadelphia, PA, USA.
FAU - Diamond, Scott L
AU  - Diamond SL
LA  - eng
GR  - R01 HL103419/HL/NHLBI NIH HHS/United States
GR  - R01 HL-103419/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - England
TA  - Biotechniques
JT  - BioTechniques
JID - 8306785
RN  - 0 (Annexin A5)
RN  - 0 (Crotalid Venoms)
RN  - 0 (Lectins, C-Type)
RN  - 0 (Oligopeptides)
RN  - 0 (P-Selectin)
RN  - 0 (Peptide Fragments)
RN  - 0 (Phosphatidylserines)
RN  - 0 (alanyl-tyrosyl-prolyl-glycyl-lysyl-phenylalanine)
RN  - 0 (thrombin receptor peptide (42-47))
RN  - 37206-04-5 (convulxin)
RN  - 61D2G4IYVH (Adenosine Diphosphate)
RN  - 76898-47-0 (15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid)
RN  - K7Q1JQR04M (Dinoprostone)
SB  - IM
MH  - 15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5,13-dienoic Acid/pharmacology
MH  - Adenosine Diphosphate/pharmacology
MH  - Annexin A5/chemistry/metabolism
MH  - Blood Donors
MH  - Blood Platelets/chemistry/cytology/*drug effects/metabolism
MH  - Cluster Analysis
MH  - Crotalid Venoms/pharmacology
MH  - Dinoprostone/pharmacology
MH  - Flow Cytometry/*methods
MH  - Humans
MH  - Lectins, C-Type
MH  - Male
MH  - Oligopeptides/pharmacology
MH  - P-Selectin/chemistry/metabolism
MH  - Peptide Fragments/pharmacology
MH  - Phenotype
MH  - Phosphatidylserines
MH  - Precision Medicine
MH  - Signal Transduction
PMC - PMC4086368
MID - NIHMS599833
COIS- Competing Interests The authors declare no competing interests.
EDAT- 2013/05/15 06:00
MHDA- 2013/11/16 06:00
PMCR- 2014/07/08
CRDT- 2013/05/14 06:00
PHST- 2013/01/29 00:00 [received]
PHST- 2013/04/22 00:00 [accepted]
PHST- 2013/05/14 06:00 [entrez]
PHST- 2013/05/15 06:00 [pubmed]
PHST- 2013/11/16 06:00 [medline]
PHST- 2014/07/08 00:00 [pmc-release]
AID - 000114027 [pii]
AID - 10.2144/000114027 [doi]
PST - ppublish
SO  - Biotechniques. 2013 May;54(5):271-7. doi: 10.2144/000114027.