PR3 特异性 B 细胞及其免疫球蛋白序列的分离和鉴定。

Isolation and characterisation of PR3-specific B cells and their immunoglobulin sequences.

机构信息

Garvan Institute of Medical Research, UNSW Sydney, 384 Victoria Street, Darlinghurst, NSW, 2010, Australia; St Vincent's Clinical School, Faculty of Medicine, UNSW Sydney, NSW, 2052, Australia.

出版信息

J Autoimmun. 2024 Jan;142:103129. doi: 10.1016/j.jaut.2023.103129. Epub 2023 Nov 11.

DOI:10.1016/j.jaut.2023.103129

PMID:37952292

Abstract

BACKGROUND

PR3 autoantibodies are essential to the diagnosis and monitoring of granulomatosus with polyangiitis, but to date no PR3 autoantibody sequences have been published.

OBJECTIVES

To identify and characterise PR3-specific B cells from the peripheral blood of patients with PR3 autoantibodies.

METHODS

Peripheral blood mononuclear cells from seven patients with PR3 autoantibodies were stained with PR3. B cells that bound PR3 underwent single cell sorting, transcriptome sequencing, and their immunoglobulin sequences expressed as antibodies and tested for PR3-specificity by ELISA.

RESULTS

We identified 19 PR3-specific B cells from only one PR3-seropositive patient at a low frequency (0.0075 % of B cells) in the peripheral blood. These were polyclonal, IgG and enriched for IgG4, lambda pairing, IGHJ6 gene usage, CDRH3 length, IGHE and CD71 expression. They demonstrated relatively low levels of somatic hypermutation and variably reduced PR3 binding when reverted to germline.

CONCLUSIONS

Identifying PR3-specific B cells in the peripheral blood is possible but challenging and those we did identify exhibited features suggesting that PR3-self reactivity may occur early in B-cell development.

摘要

背景

PR3 自身抗体对于肉芽肿性多血管炎的诊断和监测至关重要，但迄今为止，尚未发表 PR3 自身抗体的序列。

目的

从 PR3 自身抗体患者的外周血中鉴定和表征 PR3 特异性 B 细胞。

方法

用 PR3 对 7 名 PR3 自身抗体患者的外周血单个核细胞进行染色。与 PR3 结合的 B 细胞进行单细胞分选、转录组测序，并将其免疫球蛋白序列表达为抗体，通过 ELISA 测试其对 PR3 的特异性。

结果

我们仅从一名 PR3 血清阳性患者的外周血中以低频率（B 细胞的 0.0075%）鉴定出 19 个 PR3 特异性 B 细胞。这些 B 细胞是多克隆的，IgG 型，富含 IgG4、lambda 配对、IGHJ6 基因使用、CDRH3 长度、IGHE 和 CD71 表达。它们表现出相对较低水平的体细胞超突变，当恢复为原始状态时，PR3 结合能力可变降低。