Identification of B cell defects using age-defined reference ranges for in vivo and in vitro B cell differentiation

Primary immunodeficiencies consist to a large extent of B cell defects, as indicated by inadequate Ab levels or response upon immunization. Many B cell defects have not yet been well characterized. Our objective was to create reliable in vivo and in vitro assays to routinely analyze human B cell dif...

Full description

Saved in:
Bibliographic Details
Published in:The Journal of immunology (1950) 2013-05, Vol.190 (10), p.5012-5019
Main Authors: aan de Kerk, Daan J, Jansen, Machiel H, ten Berge, Ineke J M, van Leeuwen, Ester M M, Kuijpers, Taco W
Format: Article
Language:English
Subjects:
ADP-ribosyl Cyclase 1 - analysis
Adult
Antibodies - blood
Antigens, CD20 - analysis
B-Lymphocyte Subsets - classification
B-Lymphocyte Subsets - cytology
B-Lymphocyte Subsets - immunology
B-Lymphocytes - classification
B-Lymphocytes - cytology
B-Lymphocytes - immunology
Cell Differentiation - immunology
Cell Proliferation
Cells, Cultured
Common Variable Immunodeficiency - classification
Common Variable Immunodeficiency - immunology
Female
Humans
Immunoglobulins - biosynthesis
Immunophenotyping
Lymphocyte Activation - immunology
Male
Reference Values
Syndecan-1 - analysis
T-Lymphocytes - immunology
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Primary immunodeficiencies consist to a large extent of B cell defects, as indicated by inadequate Ab levels or response upon immunization. Many B cell defects have not yet been well characterized. Our objective was to create reliable in vivo and in vitro assays to routinely analyze human B cell differentiation, proliferation, and Ig production and to define reference ranges for different age categories. The in vitro assays were applied to classify the developmental and/or functional B cell defects in patients previously diagnosed with common variable immunodeficiency. Apart from standard immunophenotyping of circulating human B cell subsets, an in vitro CFSE dilution assay was used for the assessment of proliferative capacity comparing T cell-dependent and T cell-independent B cell activation. Plasmablast/plasma cell differentiation was assessed by staining for CD20, CD38, and CD138, and measurement of in vitro Ig secretion. At young age, B cells proliferate upon in vitro activation, but neither differentiate nor produce IgG. These latter functions reached adult levels at 5 and 10 y of age for T cell-dependent versus T cell-independent stimulations, respectively. The capacity of B cells to differentiate into plasmablasts and to produce IgG appeared to be contained within the switched memory B cell pool. Using these assays, we could categorize common variable immunodeficiency patients into subgroups and identified a class-switch recombination defect caused by an UNG mutation in one of the patients. We defined age-related reference ranges for human B cell differentiation. Our findings indicate that in vivo B cell functionality can be tested in vitro and helps to diagnose suspected B cell defects.
ISSN:0022-1767
1550-6606
DOI:10.4049/jimmunol.1201807