Human CD19 and CD40L deficiencies impair antibody selection and differentially affect somatic hypermutation

doi:10.1016/j.jaci.2013.11.015

Journal of Allergy and Clinical Immunology

Volume 134, Issue 1, July 2014, Pages 135-144.e7

https://doi.org/10.1016/j.jaci.2013.11.015 Get rights and content

Background

Individuals with genetic defects in CD40 ligand (CD40L) or B-cell antigen receptor coreceptor molecules CD19 and CD81 suffer from an antibody deficiency. Still, these patients carry low levels of memory B cells and serum antibodies.

Objective

We sought to assess why the remaining memory B cells and antibodies in the blood of these patients do not provide functional immunity.

Methods

We included CD19-deficient patients (n = 8), CD40L-deficient patients (n = 8), and healthy controls (n = 50) to perform detailed flow cytometry on blood B cells, molecular analysis of IgA and IgG transcripts, as well as functional analysis of B-cell activation.

Results

CD19-deficient and CD40L-deficient patients carried reduced numbers of all memory B-cell subsets except CD27⁻IgA⁺ B cells. Their immunoglobulin heavy chain class-switched transcripts contained less somatic mutations and reduced usage of IgM-distal IgG2 and IgA2 subclasses. The selection strength of mutations for antigen binding was significantly lower than in controls, whereas selection to maintain superantigen binding was normal. Furthermore, the patients showed impaired selection against inherently autoreactive properties of their immunoglobulins. Somatic hypermutation analysis revealed decreased activation-induced cytidine deaminase and uracil-DNA glycosylase 2 activity in CD40L deficiency and increased uracil-DNA glycosylase 2 but decreased mismatch repair in CD19 deficiency. B-cell activation studies revealed that this was at least in part due to transcriptional regulation of DNA repair genes.

Conclusions

This study on CD19 and CD40L deficiencies illustrates that both the B-cell antigen receptor and CD40 signaling pathways are required for the selection of immunoglobulin reactivity. Still, they differentially mediate DNA repair pathways during somatic hypermutation, thereby together shaping the human in vivo antigen-experienced B-cell repertoire.

Section snippets

Patients

Eight CD19-deficient and 8 CD40L-deficient patients suffering from an antibody deficiency were included in this study (see Table E1 in this article's Online Repository at www.jacionline.org). Seven CD19-deficient patients had biallelic genetic defects in the CD19 gene, and 1 patient had a homozygous mutation in the CD81 gene.17, 18, 19, 23, 24 Eight male patients suffered from a hyper-IgM syndrome because of the absence of CD40L expression on activated T cells and carried hemizygous mutations

Memory B-cell subsets in CD19 and CD40L deficiencies

To study the extent of impaired humoral immunity in CD19-deficient and CD40L-deficient patients (Table E1), we performed extensive flow cytometric immunophenotyping of their blood B-cell compartments. CD40L-deficient patients showed a nearly complete lack of primary (CD27⁺IgM-only and CD27⁻IgG⁺) and secondary GC-derived (CD27⁺IgG⁺ and CD27⁺IgA⁺) memory B cells, but had clearly detectable CD27⁺IgM⁺IgD⁺ “natural effector” B cells and normal numbers of CD27⁻IgA⁺ B cells (see Fig E1 in this

Discussion

Here, we analyzed antibody maturation in patients with CD19 and CD40L deficiencies to understand the absence of functional B-cell immunity in these patients. Although selection for superantigen binding was normal in CD19 deficiency, both patient groups showed decreased SHM and reduced selection strengths for antigen binding. Furthermore, selection against inherently autoreactive IGHV genes and IGH-CDR3 properties were impaired. Interestingly, SHM targeting and repair were differentially

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: This work was supported by a grant from Erasmus University Rotterdam (EUR-Fellowship) to M.C.v.Z., and ZonMW Vidi grant 015.008.046 to M.v.d.B.

: Disclosure of potential conflict of interest: M. C. van Zelm has received research support from Erasmus University Rotterdam (EUR) in the form of a fellowship, ZonMW, Sophia Children's Hospital Fund, and ACTA Amsterdam. G. J. Driessen has received research support from EUR in the form of a fellowship and has received the Baxter Grant. F. Mascart has received research support from the ECFP-7 program and FRSM. L. Boon has received consultancy fees from Fast Forward Therapeutics; is employed by Bioceros; and has stock/stock options in Fast Forward Therapeutics, Bioceros, Biocerox, and Broteio. M. van der Burg has received research support from ZonMW (Vidi grant no. 91712312). The rest of the authors declare that they have no relevant conflicts of interest.

View full text

Immune deficiencies, infection, and systemic immune disordersHuman CD19 and CD40L deficiencies impair antibody selection and differentially affect somatic hypermutation

Background

Objective

Methods

Results

Conclusions

Section snippets

Patients

Memory B-cell subsets in CD19 and CD40L deficiencies

Discussion

Trends Immunol

Semin Immunol

Cell

Blood

Immunity

Cell

Cell

Immunity

Blood

Immunol Today

Cell

J Biol Chem

Blood

Curr Biol

Ordered rearrangement of immunoglobulin heavy chain variable region segments

Embo J

Evolutionary and somatic selection of the antibody repertoire in the mouse

Science

Mechanism and regulation of class switch recombination

Annu Rev Immunol

Immunoglobulin class-switch DNA recombination: induction, targeting and beyond

Nat Rev Immunol

Primary antibody deficiencies

Nat Rev Immunol

CD40-CD40L independent Ig gene hypermutation suggests a second B cell diversification pathway in humans

Proc Natl Acad Sci U S A

Viral induction of AID is independent of the interferon and the Toll-like receptor signaling pathways but requires NF-kappaB

J Exp Med

CpG DNA induces IgG class switch DNA recombination by activating human B cells through an innate pathway that requires TLR9 and cooperates with IL-10

J Immunol

The transmembrane activator TACI triggers immunoglobulin class switching by activating B cells through the adaptor MyD88

Nat Immunol

An antibody-deficiency syndrome due to mutations in the CD19 gene

N Engl J Med

CD81 gene defect in humans disrupts CD19 complex formation and leads to antibody deficiency

J Clin Invest

Immune deficiencies, infection, and systemic immune disorders
Human CD19 and CD40L deficiencies impair antibody selection and differentially affect somatic hypermutation