Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2020, 164(3):247-254 | DOI: 10.5507/bp.2019.052

Multiparametric flow cytometry analysis of peripheral blood B cell trafficking differences among Epstein-Barr virus infected and uninfected subpopulations

Katerina Zachova^a, Petr Kosztyu^a, Josef Zadrazil^b, Karel Matousovic^c, Karel Vondrak^d, Petr Hubacek^e, Klara Kostovcikova^h, Helena Tlaskalova Hogenova^h, Jiri Mestecky^f,g,h, Milan Raska^a,g

^a Department of Immunology, Faculty of Medicine and Dentistry, Palacky University Olomouc and University Hospital Olomouc, Czech Republic

^b Department of Internal Medicine III - Nephrology, Rheumatology and Endocrinology, Palacky University Olomouc and University Hospital Olomouc, Czech Republic

^c Department of Internal Medicine, 2

^nd Faculty Medicine, Charles University in Prague and University Hospital Motol, Prague, Czech Republic

^d Department of Pediatrics, 2

^e Department of Microbiology, 2

^f Department of Medicine, University of Alabama at Birmingham, Birmingham, Alabama, USA

^g Department of Microbiology, University of Alabama at Birmingham, Birmingham, Alabama, USA

^h Laboratory of Cellular and Molecular Immunology, Institute of Microbiology, Czech Academy of Sciences, Prague, Czech Republic

Aims: Epstein-Barr virus (EBV) targets predominantly B cells and these cells could acquire new phenotype characteristics. Here we analyzed whether EBV-infected and -uninfected B cells from healthy subjects differ in proportion of dominant phenotypes, maturation stage, and homing receptors expression.

Methods: EBV-infected and -uninfected cells were identified by flow cytometry using fluorophore-labeled EBV RNA-specific DNA probes combined with fluorophore-labeled antibody to surface lineage markers, integrins, chemokine receptors, and immunoglobulin isotypes, including intracellular ones.

Results: Our results show that the trafficking characteristics of EBER^pos B cells are distinct from EBER^neg B cells with most dominant differences detected for α4β1 and α4β7 and CCR5 and CCR7. EBV-positive cells are predominantly memory IgM⁺ B cells or plasmablasts/plasma cells (PB/PC) positive for IgA or less for IgM. In comparison to uninfected B cells, less EBV-positive B cells express α4β7 and almost no cells express α4β1. EBV-positive B cells contained significantly higher proportion of CCR5⁺ and CCR7⁺ cells in comparison to EBV-negative cells. In vitro exposure of blood mononuclear cells to pro-inflammatory cytokine IL-6 reduces population of EBV-positive B cell.

Conclusion: Although EBV-infected B cells represent only a minor subpopulation, their atypical functions could contribute in predisposed person to development abnormities such as some autoimmune diseases or tumors. Using multi-parameter flow cytometry we characterized differences in migration of EBV-positive and -negative B cells of various maturation stage and isotype of produced antibodies particularly different targeting to mucosal tissues of gastrointestinal and respiratory tracts.

Keywords: Epstein-Barr virus, cell trafficking, IgM, IgA, naïve B cells, memory B cells, plasma blast, plasma cell

Received: May 20, 2019; Revised: June 24, 2019; Accepted: June 27, 2019; Prepublished online: October 12, 2019; Published: September 17, 2020  Show citation