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Front. Immunol. | doi: 10.3389/fimmu.2019.01271

EUROFLOW-BASED FLOWCYTOMETRIC DIAGNOSTIC SCREENING AND CLASSIFICATION OF PRIMARY IMMUNODEFICIENCIES OF THE LYMPHOID SYSTEM

  • 1Leiden University Medical Center, Netherlands
  • 2Charles University, Czechia
  • 3Institute of Molecular and Cellular Cancer Biology (IBMCC), Spain
  • 4Instituto de Investigación Biomédica de Salamanca (IBSAL), Spain
  • 5Masaryk University, Czechia
  • 6St. Anne's University Hospital Brno, Czechia
  • 7University Hospital La Paz, Spain
  • 8Erasmus Medical Center, Erasmus University Rotterdam, Netherlands
  • 9University of Oxford, United Kingdom
  • 10Ghent University Hospital, Belgium
  • 11Faculdade de Medicina, Universidade de Lisboa, Portugal
  • 12Monash University, Australia

Guidelines for screening for primary immunodeficiencies (PID) are well-defined and several consensus diagnostic strategies have been proposed. These consensus proposals have only partially been implemented due to lack of standardization in laboratory procedures, particularly in flowcytometry. The main objectives of the EuroFlow Consortium were to innovate and thoroughly standardize the flowcytometric techniques and strategies for reliable and reproducible diagnosis and classification of PID of the lymphoid system.
The proposed EuroFlow antibody panels comprise one orientation tube and seven classification tubes and corresponding databases of normal and PID samples. The 8-color 12-antibody PID Orientation tube (PIDOT) aims at identification and enumeration of the main lymphocyte and leukocyte subsets; this includes naïve pre-germinal center (GC) and antigen-experienced post-GC memory B-cells and plasmablasts. The seven additional 8(-12)-color tubes can be used according to the EuroFlow PID algorithm in parallel or subsequently to the PIDOT for more detailed analysis of B-cell and T-cell subsets to further classify PID of the lymphoid system. The Pre-GC, Post-GC, and immunoglobulin heavy chain (IgH)-isotype B-cell tubes aim at identification and enumeration of B-cell subsets for evaluation of B-cell maturation blocks and specific defects in IgH-subclass production. The severe combined immunodeficiency (SCID) tube and T-cell memory/effector subset tube aim at identification and enumeration of T-cell subsets for assessment of T-cell defects, such as SCID. In case of suspicion of antibody deficiency, PIDOT is preferably directly combined with the IgH isotype tube(s) and in case of SCID suspicion (e.g. in newborn screening programs) the PIDOT is preferably directly combined with the SCID T-cell tube.
The proposed ≥8-color antibody panels and corresponding reference databases combined with the EuroFlow PID algorithm are designed to provide fast, sensitive and cost-effective flowcytometric diagnosis of PID of the lymphoid system, easily applicable in multicenter diagnostic settings world-wide.

Keywords: immunodeficiency, Immunophenotyping, Flowcytometry, diagnosis, classification‐, EuroFlow, standardization

Received: 06 Dec 2018; Accepted: 17 May 2019.

Edited by:

Sergio Rosenzweig, National Institutes of Health (NIH), United States

Reviewed by:

Thomas A. Fleisher, American Academy of Allergy, Asthma and Immunology, United States
Vijaya Knight, School of Medicine, University of Colorado Denver, United States  

Copyright: © 2019 Dongen, van der Burg, Kalina, Perez-Andres, Mejstrikova, Vlkova, Lopez-Granados, Wentink, Kienzler, Philippe, Sousa, van Zelm, Blanco and Orfao. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence: Prof. Jacques J.M. V. Dongen, Leiden University Medical Center, Leiden, Netherlands, j.j.m.van_dongen@lumc.nl