肾-骨髓联合移植诱导免疫耐受患者外周血B淋巴细胞重构的动态特点及其意义

doi:10.13481/j.1671-587x.20180223

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (02): 332-338.doi: 10.13481/j.1671-587x.20180223

肾-骨髓联合移植诱导免疫耐受患者外周血B淋巴细胞重构的动态特点及其意义

张丽鹏¹, 荣春书², 谷一鸣¹, 傅耀文¹, 高宝山¹

1. 吉林大学第一医院泌尿系统疾病诊治中心, 吉林长春 130021;
2. 长春中医药大学附属医院脑病科, 吉林长春 130021

收稿日期:2017-10-09 出版日期:2018-03-28 发布日期:2018-03-30
通讯作者: 高宝山,副教授,硕士研究生导师(Tel:0431-81875825,E-mail:gaobaoshan@medmail.com.cn);傅耀文,教授,博士研究生导师(Tel:0431-88782905,E-mail:fuyaowen@medmail.com.cn) E-mail:gaobaoshan@medmail.com.cn;fuyaowen@medmail.com.cn
作者简介:张丽鹏(1990-),男,山西省晋中市人,在读医学硕士,主要从事泌尿系肿瘤和肾移植方面的研究。
基金资助:
国家自然科学基金资助课题（81501381）；吉林省科技厅科技发展计划项目资助课题（20160101100JC）

Dynamicfeatures of B lymphocyte reconstruction in peripheral blood of patients with immune tolerance induced with combined transplantation of kidney and bone marrow

ZHANG Lipeng¹, RONG Chunshu², GU Yiming¹, FU Yaowen¹, GAO Baoshan¹

1. Diagnosis and Trentment Center of Urinary System Disease, First Hospital, Jilin University, Changchun 130021, China;
2. Department of Neurology, Affiliated Hospital, Changchun University of Chinese Medicine, Changchun 130021, China

Received:2017-10-09 Online:2018-03-28 Published:2018-03-30

摘要/Abstract

摘要： 目的：探讨外周血B淋巴细胞在肾-骨髓联合移植诱导免疫耐受患者中重构的动态特点，为进一步阐明B淋巴细胞在器官移植免疫耐受建立中发挥的作用提供实验依据。方法：5例尿毒症患者（患者编码分别为Pt.A、Pt.B、Pt.C、Pt.D和Pt.E）应用诱导治疗后行肾-骨髓联合移植，术后免疫抑制剂逐渐撤除。应用流式细胞术检测患者移植术前及术后外周血B淋巴细胞数量及其亚群分布，比较不同时间点B淋巴细胞的动态变化，并利用下一代基因测序技术分析免疫球蛋白重链可变区（IGHV）的分布状态。结果：除1例患者（编码为Pt.C）移植肾早期丧失功能，其他4例患者纳入研究。Pt.A、Pt.B和Pt.D均在术后1年左右外周血B细胞计数基本恢复术前水平，Pt.E外周血B淋巴细胞重构延迟。在B淋巴细胞重构过程中，伴随有高频度的CD20+CD24^high CD38^high过渡性B淋巴细胞出现，并表现为多样性的细胞克隆图谱。移植术后约6个月时患者外周血均检测出CD20+CD27+记忆性B淋巴细胞的优势亚群。通过计算术后各时点IGHV多样性指数发现，所有肾-骨髓联合移植患者术后182d时点的IGHV基因序列的香农指数（SDI）明显低于术前（P=0.004），术后1年左右逐渐恢复。不同时点B细胞IGHV基因序列的体细胞突变检测，除了Pt.B，其他患者IGHV基因序列的体细胞突变率在术后182d时点明显升高（P=0.032）。结论：肾-骨髓联合移植患者外周血中存在体细胞突变的记忆性B淋巴细胞，此类B细胞亚群在免疫耐受诱导中发挥潜在作用。

关键词: 免疫耐受, 肾脏移植, 移植物存活, B淋巴细胞, 骨髓移植

Abstract: Objective:To study the dynamic characteristics of reconstruction of B lymphocytes in peripheral blood at different time points in the patients with immune tolerance induced by combined transplantation of kidney and bone marrow,and to provide the basis for the further illumination of the role of B lymphocytes in immune tolerance of organ transplantation. Methods: Five patients (Pt.A,Pt.B,Pt.C,Pt.D,and Pt.E)with end stage of kidney disease received kidney and bone marrow transplantation after induction therapy.Immunosuppression was discontinued gradually.The number of B lymphocytes and the distribution of subsets of B lymphocytes in peripheral blood were detected by flow cytometry in the patients before and after transplantation,and the dynamic changes of B lymphocytes at different time points were compared.The distribution of immunoglobulin heavy chain variable region(IGHV) was analyzed using next generation sequencing technology. Results: Four patients were enrolled in the study expect one patient with graft failure(Pt.C).The B lymphocyte counts of Pt.A,Pt.B and Pt.D recovered approximately 1 year after transplantation and Pt.E experienced delayed reconstitution.The B lymphocyte recovery was accompanied by a high frequency of CD20+CD24^high CD38^high transitional B lymphocytes and a diversified clonal repertoire.All patients showed the prevalence of CD20+CD27+ memory B cells around 6 months after transplantation.Through the calculation of Shannon diversity index (SDI) of IGHV at each time point after transplantation,the SDI of IGHV for all transplant recipients at 182 d after transplantation was significantly lower than before transplantation (P=0.004),and this index was gradually recovered at approximately 1 year after transplantation.The calculation of somatic mutation of B lymphocyte IGHV gene sequence at different time points showed that the somatic mutation rate was elevated at 182 d for all patients except Pt.B (P=0.032). Conclusion: The presence of mutated memory B lymphocytes in peripheral blood of the patients with kidney and bone marrow transplantation is found,and the B lymphocytes play the potential contribution to tolerance induction.

Key words: immune tolerance, graft survival, kidney transplantation, B lymphocytes, bone marrow transplantation

中图分类号:

R392.4

张丽鹏, 荣春书, 谷一鸣, 傅耀文, 高宝山. 肾-骨髓联合移植诱导免疫耐受患者外周血B淋巴细胞重构的动态特点及其意义[J]. 吉林大学学报(医学版), 2018, 44(02): 332-338.

ZHANG Lipeng, RONG Chunshu, GU Yiming, FU Yaowen, GAO Baoshan. Dynamicfeatures of B lymphocyte reconstruction in peripheral blood of patients with immune tolerance induced with combined transplantation of kidney and bone marrow[J]. Journal of Jilin University Medicine Edition, 2018, 44(02): 332-338.

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肾-骨髓联合移植诱导免疫耐受患者外周血B淋巴细胞重构的动态特点及其意义

Dynamicfeatures of B lymphocyte reconstruction in peripheral blood of patients with immune tolerance induced with combined transplantation of kidney and bone marrow

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