Classification of common variable immunodeficiencies using flow cytometry and a memory B-cell functionality assay - 06/01/15
, Sybill Thomas, ScD a, ∗Abstract |
Background |
The population of patients with common variable immunodeficiency (CVID) comprises a heterogeneous group of patients with different causes of hypogammaglobulinemia predisposing to recurrent infections, higher incidence of autoimmunity, and malignancy. Although memory B cells (memBcs) are key players in humoral defense and their numbers are commonly reduced in these patients, their functionality is not part of any current classification.
Objective |
We established and validated a memBc enzyme-linked immunosorbent spot (ELISpot) assay that reveals the capacity of memBcs to develop into antibody-secreting cells and present an idea for a new classification based on this functional capacity.
Methods |
The memBc ELISpot assay, combined with flow cytometry, was applied to patients with confirmed CVID in comparison with age-matched healthy control subjects.
Results |
Ex vivo frequency of IgG-, IgM-, and IgA-secreting plasmablasts was significantly diminished by 27.2-, 2.4-, and 23.3-fold, respectively, compared with that seen in healthy control subjects. Moreover, in vitro differentiation of memBcs into antibody-secreting cells was 6.1-, 2.6-, and 3.7-fold significantly reduced for IgG-, IgM-, and IgA-secreting cells, respectively. Proliferation of memBcs correlates inversely to immunoglobulin-secreting capacity, suggesting compensatory hyperproliferation. Furthermore, patients with no serum IgA can still have a detectable IgA ELISpot assay result in vitro. Most importantly, the large heterogeneity of memBc function in patients with CVID homogenously grouped by means of fluorescence-activated cell sorting allowed additional subclassification based on memBc/plasmablast function.
Conclusion |
These data suggest almost normal memBc/immunoglobulin-secreting plasmablast functionality in some patients if sufficient stimulatory signals are delivered, which might open up opportunities for new therapeutic approaches.
Il testo completo di questo articolo è disponibile in PDF.Key words : Memory B cell, enzyme-linked immunosorbent spot assay, common variable immunodeficiency subtyping, flow cytometry, antibody-secreting cells
Abbreviations used : ASC, CVID, ELISpot, FACS, HC, Ig-sPb, memBc, PB, PE
Mappa
| Supported by BMWi-ZIM (Federal Ministry of Economics and Technology), Cooperation Project KF2088107FR9, for the grant to partially finance this project. |
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| Disclosure of potential conflict of interest: A. L. Rösel has received research support from the BMWI-ZIM (Federal Ministry of Economics and Technology) Cooperation Project. C. Scheibenbogen has consultant arrangements with CureVac, Baxter, and Biotest; is employed by Charité; has received research support from Deutsche Forschungsgemeinschaft, EFRE, Baxter, and Behring; has received payment for lectures from Baxter; and has received travel support from Octapharma and Behring. H. von Bernuth has received research support from Deutsche Forschungsgemeinschaft and Bundesministerium für Bildung und Forschung and has received payment for lectures from CSL Behring. K. Warnatz has received payment for lectures from Baxter, GlaxoSmithKline, CSL Behring, Pfizer, Biotest, Novartis Pharma, Stallergenes AG, Roche, Meridian Health Communications, Octapharma, and the American Academy of Allergy, Asthma & Immunology; has received payment for manuscript preparation from UCB Pharma; and has received payment for development of educational presentations from the European Society for Immunodeficiencies. H.-D. Volk has received kits for enzyme-linked immunosorbent spot (ELISpot) assay free of charge from AID; has consultant arrangements with Bayer, Boehringer, Novo Nordisk, CRO, and Charité; has received research support from Bayer, Novo Nordisk, and AID; and has received license fees for patents. S. Thomas has received research support from the BMWI-ZIM (the Federal Ministry of Economics and Technology) Cooperation Project; is employed by Federal Joint Committee, PSF 120606, D-10596; and has received payment for lectures from Bundeskongress Pathologie Berlin. The rest of the authors have declared that they have no relevant conflicts of interest. |
Vol 135 - N° 1
P. 198 - gennaio 2015 Ritorno al numero
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