J Neurogastroenterol Motil  
Maturation Phenotype of Peripheral Blood Monocyte/Macrophage After Stimulation with Lipopolysaccharides in Irritable Bowel Syndrome
Oscar A Rodríguez-Fandiño,1,2 Joselín Hernández-Ruiz,1 Yolanda López-Vidal,3 Luis Charúa-Guindic,4 Galileo Escobedo,1 and Max J Schmulson1,*
1Laboratorio de Hígado, Páncreas y Motilidad (HIPAM), Unidad de Investigación en Medicina Experimental, Facultad de Medicina-Universidad Nacional Autónoma de México (UNAM), Hospital General de México, Dr. Eduardo Liceaga, Mexico City, Mexico; 2Dirección de Investigación, Fundación Universitaria–Unitrópico, Yopal, Colombia; 3Programa de Inmunología Molecular Microbiana, Departamento de Microbiología y Parasitología, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Mexico City, Mexico; and 4Unidad de Coloproctología, Hospital General de México, Dr. Eduardo Liceaga, Mexico City, Mexico
Correspondence to: Max Schmulson, MD
Laboratorio de Hígado, Páncreas y Motilidad (HIPAM), Unidad de Investigación en Medicina Experimental, Facultad de Medicina, Universidad Nacional Autónoma de México (UNAM), Hospital General de México, Dr. Eduardo Liceaga, Dr. Balmis #148. Col. Doctores, C.P. 06726, México D.F., México
Tel: +52-5556232673, Fax: +52-5556232669, E-mail: maxjulio@prodigy.net.mx
Received: August 24, 2016; Accepted: October 26, 2016; Published online: January 2, 2017.
© The Korean Society of Neurogastroenterology and Motility. All rights reserved.

cc This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted noncommercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract
Background/Aims
Abnormal immune regulation and increased intestinal permeability augmenting the passage of bacterial molecules that can activate immune cells, such as monocytes/macrophages, have been reported in irritable bowel syndrome (IBS). To compare the maturation phenotype of monocytes/macrophages (CD14+) from IBS patients and controls in the presence or absence of E. coli lipopolysaccharides (LPS), in vitro.
Methods
Mononuclear cells were isolated from peripheral blood of 20 Rome II-IBS patients and 19 controls and cultured with or without LPS for 72 hours. The maturation phenotype was examined by flow cytometry as follows: M1-Early (CD11c+CD206-), M2-Advanced (CD11c-CD206+CX3CR1+); expression of membrane markers is reported as mean fluorescence intensity (MFI). The Mann-Whitney test was used and significance was set at P < 0.05
Results
In CD14+ cells, CD11c expression decreased with vs without LPS both in IBS (MFI: 8766.0 ± 730.2 vs 12 920.0 ± 949.2, P < 0.001) and controls (8233.0 ± 613.9 vs 13 750.0 ± 743.3, P < 0.001). M1-Early cells without LPS, showed lower CD11c expression in IBS than controls (MFI: 11 540.0 ± 537.5 vs 13 860.0 ± 893.7, P = 0.040), while both groups showed less CD11c in response to LPS (P < 0.01). Furthermore, the percentage of "Intermediate" (CD11c+CD206+CX3CR1+) cells without LPS, was higher in IBS than controls (IBS = 9.5 ± 1.5% vs C = 4.9 ± 1.4%, P < 0.001. Finally, fractalkine receptor (CX3CR1) expression on M2-Advanced cells 1 was increased when treated with LPS in controls but not in IBS (P = 0.0002).
Conclusions
The initial phase of monocyte/macrophage maturation appears to be more advanced in IBS compared to controls. However, the decreased CX3CR1 in patients with IBS than in controls when stimulated with LPS suggests a state of immune activation in IBS.
Keywords: CX3CR1; Fractalkine receptor; Irritable bowel syndrome; Maturation; Monocyte-macrophage


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