Classification of common variable immunodeficiencies using flow cytometry and a memory B-cell functionality assay

JOURNAL OF ALLERGY AND CLINICAL IMMUNOLOGY

卷:135

Article

Roesel, Amelia L.¹  Scheibenbogen, Carmen^1,2,3  Schliesser, Ulrike²  Sollwedel, Andre¹  Hoffmeister, Bodo³  Hanitsch, Leif³  von Bernuth, Horst⁴  Krueger, Renate⁴  Warnatz, Klaus⁵  Volk, Hans-Dieter^1,2  Thomas, Sybill¹ 

[1] Charite Univ Med Berlin, Inst Med Immunol, Berlin, Germany

[2] Berlin Brandenburg Ctr Regenerat Therapies, Berlin, Germany

[3] Charite Univ Med Berlin, Out Patients Clin Immunodeficenc, Berlin, Germany

[4] Charite Univ Med Berlin, Dept Pneumol & Immunol, Pediat Clin, Berlin, Germany

[5] Univ Freiburg, Freiburg, Germany

关键词: Memory B cell;    enzyme-linked immunosorbent spot assay;    common variable immunodeficiency subtyping;    flow cytometry;    antibody-secreting cells;   

DOI  :  10.1016/j.jaci.2014.06.022

来源: Elsevier

【 摘 要 】

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 23.3fold, respectively, compared with that seen in healthy control subjects. Moreover, invitro differentiation ofmemBcs into antibodysecreting 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 bymeans 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.

【 授权许可】

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