In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis

Claudio Karsulovic,1,2 Fabian Tempio,3,4 Mercedes Lopez,3,4 Julia Guerrero,1 Annelise Goecke1,2 1Laboratorio de Inmunomodulación Neuroendocrina, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile; 2Seccion de Reumatología, H...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Karsulovic C, Tempio F, Lopez M, Guerrero J, Goecke A
Formato:	article
Lenguaje:	EN
Publicado:	Dove Medical Press 2021
Materias:	classical intermediate non-classical monocytes phenotype stimulation Pathology RB1-214 Therapeutics. Pharmacology RM1-950
Acceso en línea:	https://doaj.org/article/267aaaf71222499e8355ab984cdcf87c
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:267aaaf71222499e8355ab984cdcf87c
record_format	dspace
spelling	oai:doaj.org-article:267aaaf71222499e8355ab984cdcf87c2021-12-02T13:53:35ZIn vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis1178-7031https://doaj.org/article/267aaaf71222499e8355ab984cdcf87c2021-01-01T00:00:00Zhttps://www.dovepress.com/in-vitro-phenotype-induction-of-circulating-monocytes-cd16-and-cd163-a-peer-reviewed-article-JIRhttps://doaj.org/toc/1178-7031Claudio Karsulovic,1,2 Fabian Tempio,3,4 Mercedes Lopez,3,4 Julia Guerrero,1 Annelise Goecke1,2 1Laboratorio de Inmunomodulación Neuroendocrina, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile; 2Seccion de Reumatología, Hospital Clínico Universidad de Chile, Universidad de Chile, Santiago, Chile; 3Laboratorio de Regulación e Inmunología del Cáncer, Facultad de Medicina, Universidad de Chile, Santiago, Chile; 4Instituto Milenio de Inmunología e Inmunoterapia, Facultad de Medicina, Universidad de Chile, Santiago, ChileCorrespondence: Annelise GoeckeSeccion de Reumatologia, Hospital Clínico Universidad de Chile, Universidad de Chile, Santos Dumont 999, Independencia, Santiago, ChileTel +56-229788000Email igoecke@hcuch.clIntroduction: CD14 (monocyte differentiation antigen, LPS binding protein – endotoxin receptor) and CD16 (FcγRIII, Low-affinity receptor for IgG) define three subpopulations of circulating monocytes with different inflammatory and phagocytic capabilities. Contradictory reports exist regarding both in vivo monocyte phenotype-disease association and response of these circulating monocytes to in vitro stimulation. We analyzed phenotypic changes in circulating monocytes when stimulated with LPS (pro-inflammatory stimulus) and IL-4 (alternative inflammatory stimulus).Methods: Mononuclear cells from nine healthy donors were extracted and studied for surface and intracellular markers using flow cytometry. PBMC were extracted using Ficoll technic and immediately analyzed using flow cytometry. Pro-inflammatory interleukin IL-1β and IL-6 were measured by intracellular cytometry. Mononuclear cells were stimulated using LPS and IL-4 as previously described. Changes against non-stimulated populations were statistically analyzed.Results: Compared to non-stimulated and IL-4 stimulated monocytes, LPS-stimulated cells display a singular pattern of markers, with higher levels of intracellular IL-1β and IL-6 directly correlating with CD14+CD163- cell frequency and diminishing membrane CD163 fluorescence. CD14+CD16- classical monocytes show greater percentage of CD163- cells upon LPS stimulation. CD86 levels on monocytes’ surface did not change with LPS or IL-4 stimulation.Conclusions and Discussion: We showed that CD14+CD16- classical monocytes display higher sensitivity to LPS stimulation, with more IL-1β and IL-6 levels than intermediate and non-classical monocytes. This subset also diminishes its CD163 levels on the membrane after LPS stimulation with a contemporary raise in CD163- cells, suggesting that classical monocytes preferentially acquire CD163- defined M1 characteristics upon in vitro LPS stimulation. Intermediate and non-classical monocytes respond with lower levels of interleukins and display surface proteins in an M2-type profile (CD163+).Keywords: classical, intermediate, non-classical, monocytes, phenotype, stimulationKarsulovic CTempio FLopez MGuerrero JGoecke ADove Medical Pressarticleclassicalintermediatenon-classicalmonocytesphenotypestimulationPathologyRB1-214Therapeutics. PharmacologyRM1-950ENJournal of Inflammation Research, Vol Volume 14, Pp 191-198 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	classical intermediate non-classical monocytes phenotype stimulation Pathology RB1-214 Therapeutics. Pharmacology RM1-950
spellingShingle	classical intermediate non-classical monocytes phenotype stimulation Pathology RB1-214 Therapeutics. Pharmacology RM1-950 Karsulovic C Tempio F Lopez M Guerrero J Goecke A In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis
description	Claudio Karsulovic,1,2 Fabian Tempio,3,4 Mercedes Lopez,3,4 Julia Guerrero,1 Annelise Goecke1,2 1Laboratorio de Inmunomodulación Neuroendocrina, Instituto de Ciencias Biomédicas, Facultad de Medicina, Universidad de Chile, Santiago, Chile; 2Seccion de Reumatología, Hospital Clínico Universidad de Chile, Universidad de Chile, Santiago, Chile; 3Laboratorio de Regulación e Inmunología del Cáncer, Facultad de Medicina, Universidad de Chile, Santiago, Chile; 4Instituto Milenio de Inmunología e Inmunoterapia, Facultad de Medicina, Universidad de Chile, Santiago, ChileCorrespondence: Annelise GoeckeSeccion de Reumatologia, Hospital Clínico Universidad de Chile, Universidad de Chile, Santos Dumont 999, Independencia, Santiago, ChileTel +56-229788000Email igoecke@hcuch.clIntroduction: CD14 (monocyte differentiation antigen, LPS binding protein – endotoxin receptor) and CD16 (FcγRIII, Low-affinity receptor for IgG) define three subpopulations of circulating monocytes with different inflammatory and phagocytic capabilities. Contradictory reports exist regarding both in vivo monocyte phenotype-disease association and response of these circulating monocytes to in vitro stimulation. We analyzed phenotypic changes in circulating monocytes when stimulated with LPS (pro-inflammatory stimulus) and IL-4 (alternative inflammatory stimulus).Methods: Mononuclear cells from nine healthy donors were extracted and studied for surface and intracellular markers using flow cytometry. PBMC were extracted using Ficoll technic and immediately analyzed using flow cytometry. Pro-inflammatory interleukin IL-1β and IL-6 were measured by intracellular cytometry. Mononuclear cells were stimulated using LPS and IL-4 as previously described. Changes against non-stimulated populations were statistically analyzed.Results: Compared to non-stimulated and IL-4 stimulated monocytes, LPS-stimulated cells display a singular pattern of markers, with higher levels of intracellular IL-1β and IL-6 directly correlating with CD14+CD163- cell frequency and diminishing membrane CD163 fluorescence. CD14+CD16- classical monocytes show greater percentage of CD163- cells upon LPS stimulation. CD86 levels on monocytes’ surface did not change with LPS or IL-4 stimulation.Conclusions and Discussion: We showed that CD14+CD16- classical monocytes display higher sensitivity to LPS stimulation, with more IL-1β and IL-6 levels than intermediate and non-classical monocytes. This subset also diminishes its CD163 levels on the membrane after LPS stimulation with a contemporary raise in CD163- cells, suggesting that classical monocytes preferentially acquire CD163- defined M1 characteristics upon in vitro LPS stimulation. Intermediate and non-classical monocytes respond with lower levels of interleukins and display surface proteins in an M2-type profile (CD163+).Keywords: classical, intermediate, non-classical, monocytes, phenotype, stimulation
format	article
author	Karsulovic C Tempio F Lopez M Guerrero J Goecke A
author_facet	Karsulovic C Tempio F Lopez M Guerrero J Goecke A
author_sort	Karsulovic C
title	In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis
title_short	In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis
title_full	In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis
title_fullStr	In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis
title_full_unstemmed	In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis
title_sort	in vitro phenotype induction of circulating monocytes: cd16 and cd163 analysis
publisher	Dove Medical Press
publishDate	2021
url	https://doaj.org/article/267aaaf71222499e8355ab984cdcf87c
work_keys_str_mv	AT karsulovicc invitrophenotypeinductionofcirculatingmonocytescd16andcd163analysis AT tempiof invitrophenotypeinductionofcirculatingmonocytescd16andcd163analysis AT lopezm invitrophenotypeinductionofcirculatingmonocytescd16andcd163analysis AT guerreroj invitrophenotypeinductionofcirculatingmonocytescd16andcd163analysis AT goeckea invitrophenotypeinductionofcirculatingmonocytescd16andcd163analysis
_version_	1718392368323887104

In vitro Phenotype Induction of Circulating Monocytes: CD16 and CD163 Analysis

Ejemplares similares