Assessment of variation in B-cell receptor heavy chain repertoire in patients with end-stage renal disease by high-throughput sequencing

Abstract Background and Aims End-stage renal disease (ESRD) is linked to immunodeficiency, which makes a significant contribution to morbidity and mortality. Disturbances in innate and adaptive immunity have been described in patients on dialysis, although their association with the therapy itself is yet to be defined. The present study aimed to assess the impact of dialysis on B cell subpopulations Method B cells (CD19+) and their subsets B1a (CD19+CD5+), naive (CD19+CD27−), memory (CD19+CD27+), (CD19+BAFFR+) and (CD19+IgM+), were quantified using flow-cytometry of in the peripheral blood of ESRD patients, the first day on dialysis (T0), and repeated 6 months later (T6). The results were compared to age-matched healthy control group. Exclusion criteria were age <18 or>75 years, active autoimmune or chronic inflammatory disease, medical history of malignancy, corticosteroids or immunosuppresive treatment for the last 12 months Results Pre dialysis ESRD patients had reduced lymphocyte count (1527±646μ/L vs. 2459±520μ/L, p<0.001) and B cell (CD19+) count (82.7±59.5μ/L vs. 177.6±73.8μ/L, p<0.001) compared to controls, whereas the percentages of B cell subsets were not particularly affected, except for B1a subset which presented a significant increase (4.1±3.6% vs. 0.7±0.7% p<0.001). In 17 patients who had a follow-up sample 6 months later, the percentage of most subsets was reduced (CD19+CD5+: 1.02±0.8% from 3.6±4.6%, p=0.015, Naive: 40±22.3% from 61±17.4%, p=0.001, CD19+BAFF+:75.8±12.6% from 82.1±9.1%, p=0.04,), apart from memory B cells percentage, which was increased (49.4±52.1% from 32,9±35,5%, p=0.01) and CD19+ IgM+ percentage, which was unaffected . Conclusion A significant reduction of almost all subsets of B cells was noticed in patients with ESRD on pre-dialysis stage. Furthermore, the initiation of renal replacement therapy may be linked to further alterations in B cells subpopulations, especially at their early stages.

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Immunoglobulin heavy chains are sufficient to determine most B cell clonal relationships1

10.1101/665760 ◽

2019 ◽

Author(s):

Julian Q. Zhou ◽

Steven H. Kleinstein

Keyword(s):

B Cell ◽

High Throughput ◽

Heavy Chain ◽

Clonal Expansion ◽

Cell Receptor ◽

B Cell Receptor ◽

Light Chains ◽

Heavy Chains ◽

Computational Inference ◽

Cell Clonal Expansion

AbstractB cell clonal expansion is vital for adaptive immunity. High-throughput B cell receptor (BCR) sequencing enables investigating this process, but requires computational inference to identify clonal relationships. This inference usually relies on only the BCR heavy chain, as most current protocols do not preserve heavy:light chain pairing. The extent to which paired light chains aids inference is unknown. Using human single-cell paired BCR datasets, we assessed the ability of heavy chain-based clonal clustering to identify clones. Of the expanded clones identified, <20% grouped cells expressing inconsistent light chains. Heavy chains from these misclustered clones contained more distant junction sequences and shared fewer V segment mutations than the accurate clones. This suggests that additional heavy chain information could be leveraged to refine clonal relationships. Conversely, light chains were insufficient to refine heavy chain-based clonal clusters. Overall, the BCR heavy chain alone is sufficient to identify clonal relationships with confidence.

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High-Throughput Sequencing of the B Cell Receptor in African Burkitt Lymphoma Reveals Clues to Pathogenesis

immuneACCESS ◽

10.21417/b7ms37 ◽

2018 ◽

Author(s):

K Lombardo ◽

D Coffey ◽

A Morales ◽

C Carlson ◽

A Towlerton ◽

...

Keyword(s):

B Cell ◽

High Throughput ◽

Burkitt Lymphoma ◽

High Throughput Sequencing ◽

Cell Receptor ◽

B Cell Receptor

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Rituximab versus cyclophosphamide in ANCA-associated renal vasculitis: 2-year results of a randomised trial

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2014-206404 ◽

2015 ◽

Vol 74 (6) ◽

pp. 1178-1182 ◽

Cited By ~ 114

Author(s):

Rachel B Jones ◽

Shunsuke Furuta ◽

Jan Willem Cohen Tervaert ◽

Thomas Hauser ◽

Raashid Luqmani ◽

...

Keyword(s):

Renal Disease ◽

B Cell ◽

End Stage Renal Disease ◽

Remission Induction ◽

Control Group ◽

Intravenous Cyclophosphamide ◽

Anca Associated Vasculitis ◽

Renal Vasculitis ◽

Stage Renal Disease ◽

End Stage

ObjectivesThe RITUXVAS trial reported similar remission induction rates and safety between rituximab and cyclophosphamide based regimens for antineutrophil cytoplasm antibody (ANCA)-associated vasculitis at 12 months; however, immunosuppression maintenance requirements and longer-term outcomes after rituximab in ANCA-associated renal vasculitis are unknown.MethodsForty-four patients with newly diagnosed ANCA-associated vasculitis and renal involvement were randomised, 3:1, to glucocorticoids plus either rituximab (375 mg/m2/week×4) with two intravenous cyclophosphamide pulses (n=33, rituximab group), or intravenous cyclophosphamide for 3–6 months followed by azathioprine (n=11, control group).ResultsThe primary end point at 24 months was a composite of death, end-stage renal disease and relapse, which occurred in 14/33 in the rituximab group (42%) and 4/11 in the control group (36%) (p=1.00). After remission induction treatment all patients in the rituximab group achieved complete B cell depletion and during subsequent follow-up, 23/33 (70%) had B cell return. Relapses occurred in seven in the rituximab group (21%) and two in the control group (18%) (p=1.00). All relapses in the rituximab group occurred after B cell return.ConclusionsAt 24 months, rates of the composite outcome of death, end-stage renal disease and relapse did not differ between groups. In the rituximab group, B cell return was associated with relapse.Trial registration numberISRCTN28528813.

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