PMID- 34987522
OWN - NLM
STAT- MEDLINE
DCOM- 20220225
LR  - 20220225
IS  - 1664-3224 (Electronic)
IS  - 1664-3224 (Linking)
VI  - 12
DP  - 2021
TI  - Tracing Human IgE B Cell Antigen Receptor-Bearing Cells With a Monoclonal 
      Anti-Human IgE Antibody That Specifically Recognizes Non-Receptor-Bound IgE.
PG  - 803236
LID - 10.3389/fimmu.2021.803236 [doi]
LID - 803236
AB  - Up to 30% of the population suffers from immunoglobulin E (IgE)-mediated 
      allergies. Despite current stepwise gating approaches, the unambiguous 
      identification of human IgE-producing cells by flow cytometry and immunohistology 
      remains challenging. This is mainly due to the scarcity of these cells and the 
      fact that IgE is not only expressed in a membrane-bound form on the surface of 
      IgE-producing cells in form of the B cell antigen receptor (BCR), but is more 
      frequently found on various cell types bound to the low and high affinity 
      receptors, CD23 and FcϵRI, respectively. Here we sought to develop a sequential 
      gating strategy for unambiguous detection of cells bearing the IgE BCR on their 
      surface. To that aim we first tested the monoclonal anti-IgE antibody omalizumab 
      for its ability to discriminate between IgE BCR and receptor-bound IgE using 
      cells producing IgE or bearing IgE bound to CD23 as well as basophils exhibiting 
      FcϵRI receptor-bound IgE. Using flow cytometry, we demonstrated that omalizumab 
      recognized IgE producing cells with a high sensitivity of up to 1 IgE(+) cell in 
      1000 human peripheral blood mononuclear cells (PBMCs). These results were 
      confirmed by confocal microscopy both in cell suspensions as well as in nasal 
      polyp tissue sections. Finally, we established a consecutive gating strategy 
      allowing the clear identification of class-switched, allergen-specific IgE(+) 
      memory B cells and plasmablasts/plasma cells in human PBMCs. Birch pollen 
      specific IgE(+) memory B cells represented on average 0.734% of total CD19(+) B 
      cells in allergic patients after allergen exposure. Thus, we developed a new 
      protocol for exclusive staining of non-receptor bound allergen-specific IgE(+) B 
      cell subsets in human samples.
CI  - Copyright (c) 2021 Zghaebi, Byazrova, Flicker, Villazala-Merino, Campion, Stanek, 
      Tu, Breiteneder, Filatov, Khaitov, Niederberger-Leppin, Eckl-Dorna and Valenta.
FAU - Zghaebi, Mohammed
AU  - Zghaebi M
AD  - Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
FAU - Byazrova, Maria
AU  - Byazrova M
AD  - National Research Centre (NRC) Institute of Immunology, Federal 
      Medical-Biological Agency (FMBA) of Russia, Moscow, Russia.
AD  - Department of Immunology, Faculty of Biology, Lomonosov Moscow State University, 
      Moscow, Russia.
FAU - Flicker, Sabine
AU  - Flicker S
AD  - Division of Immunopathology, Department of Pathophysiology and Allergy Research, 
      Center for Pathophysiology, Infectiology and Immunology, Medical University of 
      Vienna, Vienna, Austria.
FAU - Villazala-Merino, Sergio
AU  - Villazala-Merino S
AD  - Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
FAU - Campion, Nicholas J
AU  - Campion NJ
AD  - Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
FAU - Stanek, Victoria
AU  - Stanek V
AD  - Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
FAU - Tu, Aldine
AU  - Tu A
AD  - Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
FAU - Breiteneder, Heimo
AU  - Breiteneder H
AD  - Division of Medical Biotechnology, Department of Pathophysiology and Allergy 
      Research, Center for Pathophysiology, Infectiology and Immunology, Medical 
      University of Vienna, Vienna, Austria.
FAU - Filatov, Alexander
AU  - Filatov A
AD  - National Research Centre (NRC) Institute of Immunology, Federal 
      Medical-Biological Agency (FMBA) of Russia, Moscow, Russia.
AD  - Department of Immunology, Faculty of Biology, Lomonosov Moscow State University, 
      Moscow, Russia.
FAU - Khaitov, Musa
AU  - Khaitov M
AD  - National Research Centre (NRC) Institute of Immunology, Federal 
      Medical-Biological Agency (FMBA) of Russia, Moscow, Russia.
AD  - Immunology Department, Pirogov Russian National Research Medical University, 
      Moscow, Russia.
FAU - Niederberger-Leppin, Verena
AU  - Niederberger-Leppin V
AD  - Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
FAU - Eckl-Dorna, Julia
AU  - Eckl-Dorna J
AD  - Department of Otorhinolaryngology, Medical University of Vienna, Vienna, Austria.
FAU - Valenta, Rudolf
AU  - Valenta R
AD  - National Research Centre (NRC) Institute of Immunology, Federal 
      Medical-Biological Agency (FMBA) of Russia, Moscow, Russia.
AD  - Division of Immunopathology, Department of Pathophysiology and Allergy Research, 
      Center for Pathophysiology, Infectiology and Immunology, Medical University of 
      Vienna, Vienna, Austria.
AD  - Department of Clinical Immunology and Allergy, Sechenov First Moscow State 
      Medical University, Moscow, Russia.
AD  - Karl Landsteiner University of Health Sciences, Krems, Austria.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20211220
PL  - Switzerland
TA  - Front Immunol
JT  - Frontiers in immunology
JID - 101560960
RN  - 0 (Allergens)
RN  - 0 (Anti-Allergic Agents)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Antigens, CD19)
RN  - 0 (Antigens, Plant)
RN  - 0 (Epitopes)
RN  - 0 (Receptors, Antigen, B-Cell)
RN  - 0 (Receptors, IgE)
RN  - 2P471X1Z11 (Omalizumab)
RN  - 37341-29-0 (Immunoglobulin E)
SB  - IM
MH  - Allergens/immunology
MH  - Anti-Allergic Agents/*therapeutic use
MH  - Antibodies, Monoclonal/metabolism
MH  - Antigens, CD19/metabolism
MH  - Antigens, Plant/immunology
MH  - B-Lymphocyte Subsets/*immunology
MH  - Betula/immunology
MH  - Cell Separation
MH  - Epitopes
MH  - Flow Cytometry
MH  - Humans
MH  - Immunoglobulin Class Switching
MH  - Immunoglobulin E/*metabolism
MH  - Immunologic Memory
MH  - Omalizumab/*therapeutic use
MH  - Pollen/immunology
MH  - Protein Binding
MH  - Receptors, Antigen, B-Cell/*metabolism
MH  - Receptors, IgE/metabolism
MH  - Rhinitis, Allergic, Seasonal/*drug therapy/immunology
PMC - PMC8721004
OTO - NOTNLM
OT  - B cells
OT  - CD23
OT  - FcepsilonRI
OT  - IgE
OT  - PBMCs
OT  - allergy
OT  - omalizumab
OT  - spiking
COIS- RV has received research grants from Worg Pharmaceuticals, Hangzhou, China, HVD 
      Biotech, Vienna, Austria and Viravaxx, Vienna, Austria. He serves as a consultant 
      for Viravaxx and Worg. The remaining authors declare that the research was 
      conducted in the absence of any commercial or financial relationships that could 
      be construed as a potential conflict of interest.
EDAT- 2022/01/07 06:00
MHDA- 2022/02/26 06:00
PMCR- 2021/01/01
CRDT- 2022/01/06 05:53
PHST- 2021/10/27 00:00 [received]
PHST- 2021/11/08 00:00 [accepted]
PHST- 2022/01/06 05:53 [entrez]
PHST- 2022/01/07 06:00 [pubmed]
PHST- 2022/02/26 06:00 [medline]
PHST- 2021/01/01 00:00 [pmc-release]
AID - 10.3389/fimmu.2021.803236 [doi]
PST - epublish
SO  - Front Immunol. 2021 Dec 20;12:803236. doi: 10.3389/fimmu.2021.803236. eCollection 
      2021.