PMID- 19051238
OWN - NLM
STAT- MEDLINE
DCOM- 20090612
LR  - 20210102
IS  - 1552-4930 (Electronic)
IS  - 1552-4922 (Linking)
VI  - 75
IP  - 5
DP  - 2009 May
TI  - Suitability of various membrane lipophilic probes for the detection of 
      trogocytosis by flow cytometry.
PG  - 380-9
LID - 10.1002/cyto.a.20679 [doi]
AB  - Trogocytosis is a recently discovered phenomenon whereby lymphocytes capture 
      fragments of the plasma membrane from antigen presenting cells (APCs). Using APCs 
      labeled with widely used fluorescent lipophilic probes, we previously described a 
      trogocytosis analysis protocol (TRAP) useful to understand the mechanisms and 
      biological consequences of this process and to identify lymphocytes reacting 
      specifically with antigen-bearing APCs. We have compared the suitability of 22 
      different fluorescent lipophilic probes for use in TRAP assays with cytotoxic T 
      lymphocytes (CTL). The criteria we used were: simple and efficient incorporation 
      in APC membranes, minimal passive diffusion among cells but efficient transfer 
      onto T cells during trogocytosis. Sphingosin-based probes were found to 
      incorporate inefficiently into cells. For others with unsaturated lipid chains, 
      we found a tendency for extensive passive diffusion. In the end, about a third of 
      the probes tested were found to be suitable in TRAP assays, which all carry 
      either C16 or C18 saturated carbon chains, including some that can be excited 
      with a red laser. Moreover, we found it possible to combine TRAP assays based on 
      lipophilic probes with intracellular cytokine detection. We have identified a set 
      of new lipophilic fluorescent probes suitable for TRAP assays in combination with 
      intracellular staining.
CI  - (c) 2008 International Society for Advancement of Cytometry.
FAU - Daubeuf, Sandrine
AU  - Daubeuf S
AD  - CNRS, IPBS (Institut de Pharmacologie et de Biologie Structurale), 205 route de 
      Narbonne, Toulouse F-31077, France.
FAU - Bordier, Christine
AU  - Bordier C
FAU - Hudrisier, Denis
AU  - Hudrisier D
FAU - Joly, Etienne
AU  - Joly E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Cytometry A
JT  - Cytometry. Part A : the journal of the International Society for Analytical 
      Cytology
JID - 101235694
RN  - 0 (Fluorescent Dyes)
RN  - 82115-62-6 (Interferon-gamma)
SB  - IM
MH  - Animals
MH  - Antigen-Presenting Cells/*metabolism
MH  - Cell Line, Tumor
MH  - Cell Membrane/*metabolism
MH  - Flow Cytometry/*methods
MH  - Fluorescent Dyes/*chemistry
MH  - Interferon-gamma/metabolism
MH  - Mice
MH  - T-Lymphocytes, Cytotoxic/*metabolism
EDAT- 2008/12/04 09:00
MHDA- 2009/06/13 09:00
CRDT- 2008/12/04 09:00
PHST- 2008/12/04 09:00 [pubmed]
PHST- 2009/06/13 09:00 [medline]
PHST- 2008/12/04 09:00 [entrez]
AID - 10.1002/cyto.a.20679 [doi]
PST - ppublish
SO  - Cytometry A. 2009 May;75(5):380-9. doi: 10.1002/cyto.a.20679.