PMID- 24652220
OWN - NLM
STAT- MEDLINE
DCOM- 20141204
LR  - 20191112
IS  - 2590-7379 (Electronic)
IS  - 0120-4157 (Linking)
VI  - 33
IP  - 4
DP  - 2013 Oct-Dec
TI  - [Design of a multicolor panel to assess intracellular and surface molecules by 
      flow cytometry].
PG  - 660-72
LID - S0120-41572013000400018 [pii]
AB  - INTRODUCTION: Flow cytometry allows simultaneous detection of surface and 
      intracellular molecules on each cell. OBJECTIVE: To describe a method for 
      building up a harmonic multicolor panel with 11 flow cytometry parameters for 
      phenotypic and functional analysis on CD8 + T lymphocytes. MATERIALS AND METHODS: 
      For the multicolor panel construction, we selected the molecules and titred 
      conjugated antibodies with fluorochromes for CD3, CD8, CCR7, CD28, CD27, CD45RA, 
      CD95 and CD127 determination in peripheral blood mononuclear cells (PBMC). To 
      evaluate the panel, the conjugated antibodies were gradually added one by one and 
      fluorescence minus one (FMO) test was performed. This method was applied to 
      assess ex vivo subpopulations of T cells and the production of intracellular IFN 
      ? , IL-2 and TNF a using polyclonal stimulation with enterotoxin B from 
      Staphylococcus aureus (SEB) and antigen-specific cells with crude Trypanosoma 
      cruzi antigen. Finally, the ex vivo CD8 + T lymphocyte subpopulations frequencies 
      were analyzed in healthy individuals. RESULTS: The evaluation of the selected 
      molecules and conjugates did not show interference in the fluorescence signals 
      and detection. The frequencies of CD8 + T cells evaluated were similar to the 
      values reported in other studies. Additionally, we observed that the frequency of 
      CD8 + T lymphocytes producing IFN ? , IL-2 and TNF a was higher 6 hours after 
      culture with SEB and crude T. cruzi lysate. CONCLUSIONS: The method used for the 
      construction of a multicolor panel allows obtaining frequencies of CD8 + T 
      lymphocyte subpopulations corresponding to those reported in the literature.
FAU - Mateus, Jose
AU  - Mateus J
AD  - Grupo de Inmunobiologia y Biologia Celular, Pontificia Universidad Javeriana, 
      Bogota, D.C, Colombia.
FAU - Lasso, Paola
AU  - Lasso P
AD  - Laboratorio de Parasitologia Molecular, Pontificia Universidad Javeriana, Bogota, 
      D.C, Colombia.
FAU - Gonzalez, John Mario
AU  - Gonzalez JM
AD  - Grupo de Ciencias Basicas Medicas, Facultad de Medicina, Universidad de los 
      Andes.
FAU - Puerta, Concepcion Judith
AU  - Puerta CJ
AD  - Laboratorio de Parasitologia Molecular, Pontificia Universidad Javeriana, Bogota, 
      D.C, Colombia.
FAU - Cuellar, Adriana
AU  - Cuellar A
AD  - Grupo de Inmunobiologia y Biologia Celular, Pontificia Universidad Javeriana, 
      Bogota, D.C, Colombia.
LA  - spa
PT  - English Abstract
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
TT  - Diseno de un panel multicolor para evaluar moleculas intracelulares y de 
      superficie mediante citometria de flujo.
PL  - Colombia
TA  - Biomedica
JT  - Biomedica : revista del Instituto Nacional de Salud
JID - 8205605
RN  - 0 (Cytokines)
SB  - IM
MH  - CD8-Positive T-Lymphocytes/*chemistry
MH  - Cells, Cultured
MH  - Color
MH  - Cytokines/*analysis
MH  - Flow Cytometry/*methods
MH  - Humans
MH  - Leukocytes, Mononuclear/chemistry
MH  - T-Lymphocyte Subsets/*chemistry
EDAT- 2014/03/22 06:00
MHDA- 2014/12/15 06:00
CRDT- 2014/03/22 06:00
PHST- 2012/12/17 00:00 [received]
PHST- 2013/06/26 00:00 [accepted]
PHST- 2014/03/22 06:00 [entrez]
PHST- 2014/03/22 06:00 [pubmed]
PHST- 2014/12/15 06:00 [medline]
AID - S0120-41572013000400018 [pii]
AID - 10.7705/biomedica.v33i4.1709 [doi]
PST - ppublish
SO  - Biomedica. 2013 Oct-Dec;33(4):660-72. doi: 10.7705/biomedica.v33i4.1709.