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2.8. Flow Cytometry

MD Maria Inmaculada Dominguez-Mozo
SP Silvia Perez-Perez
NV Noelia Villarrubia
LC Lucienne Costa-Frossard
JF Jose Ignacio Fernandez-Velasco
IO Isabel Ortega-Madueño
MG Maria Angel Garcia-Martinez
EG Estefania Garcia-Calvo
HE Hector Estevez
JG Jose Luis Luque Garcia
MT Maria Josefa Torrejon
RA Rafael Arroyo
LV Luisa Maria Villar
RA Roberto Alvarez-Lafuente
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A gate including lymphocytes and monocytes and excluding debris and apoptotic cells was first established (Figure 1). CD4+ and CD8+ T cells were classified as: central memory (CM) (CCR7+ CD45RO+), naïve (CCR7+ CD45RO-), terminally differentiated (TD) (CCR7- CD45RO-) and effector memory (EM) (CCR7- CD45RO+). Regulatory CD4 T cells (Treg) were defined as CD3+ CD4+ CD25hi CD127low. B cells were classified as: plasmablasts (CD19+ CD27hi CD38hi), memory (CD19+ CD27dim CD38dim) and regulatory B cells (Breg) (CD19+ CD27- CD24hi CD38hi). Monocytes were also studied, and their subsets were classified according to the expression of CD14 and CD16 (Mo1: CD14++ CD16-; Mo2: CD14++ CD16+; Mo3: CD14+ CD16+). Dendritic cells (DCs) were differentiated into myeloid DCs (mDC: CD14- CD11c+) and plasmacytoid DCs (pDC: HLA-DR+ CD123+). Natural killer (NK) cells (CD56dim CD3-), natural killer T (NKT) cells (CD56dim CD3+) and CD56bright NK cells (CD3- CD56bright) were also studied. Data analysis was performed using FACSDiva Software V.8.0 (BD Biosciences).

Gating strategy.

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Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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