单细胞测序揭示肾移植受者抗体介导排斥反应的潜在机制

doi:10.12092/j.issn.1009-2501.2026.03.001

中国临床药理学与治疗学 ›› 2026, Vol. 31 ›› Issue (3): 289-299.doi: 10.12092/j.issn.1009-2501.2026.03.001

单细胞测序揭示肾移植受者抗体介导排斥反应的潜在机制

郑凯乐¹^,²^,³(), 付嘉钊⁴, 尤佳¹^,²^,³, 吴丹⁵, 王学彬²^,³^,*(), 王卓³

1. 蚌埠医科大学药学院，蚌埠 233030，安徽
2. 上海市儿童医院药学部，上海 200062
3. 海军军医大学第一附属医院药剂科，上海 200433
4. 海军军医大学第一附属医院器官移植科，上海 200433
5. 上海市儿童医院生物信息学中心，上海 200062

收稿日期:2025-01-07 修回日期:2025-10-14 出版日期:2026-03-26 发布日期:2026-04-03
通讯作者: 王学彬 E-mail:1604751742@qq.com;binxuewang@sjtu.edu.cn
作者简介:郑凯乐，女，硕士研究生，药师，研究方向：免疫抑制剂个体化给药。E-mail：1604751742@qq.com
基金资助:
国家自然科学基金资助项目（82173900）；促进市级医院临床技能与临床创新能力三年行动计划（SHDC2020CR4072）；2022 年上海青年药学人才能力提升项目（沪药会字[2023]04 号）；上海市儿童医院人才引进启动项目（SHSERTYY20250505）

Single-cell sequencing reveals the underlying mechanism of antibody-mediated rejection in renal transplant recipients

Kaile ZHENG¹^,²^,³(), Jiazhao FU⁴, Jia YOU¹^,²^,³, Dan WU⁵, Xuebin WANG²^,³^,*(), Zhuo WANG³

1. School of Pharmacy, Bengbu Medical College, Bengbu 233030, Anhui, China
2. Department of Pharmacy, Shanghai Children's Hospital, Shanghai 200062, China
3. Department of Pharmacy, the First Affiliated Hospital of Naval Medical University, Shanghai 200433, China
4. Department of Organ Transplantation, the First Affiliated Hospital of Naval Medical University, Shanghai 200433, China
5. Bioinformatics Center of Shanghai Children's Hospital, Shanghai 200062, China

Received:2025-01-07 Revised:2025-10-14 Online:2026-03-26 Published:2026-04-03
Contact: Xuebin WANG E-mail:1604751742@qq.com;binxuewang@sjtu.edu.cn

摘要/Abstract

摘要：

目的: 揭示肾移植受者术后发生抗体介导排斥反应（antibody-mediated rejection，ABMR）的潜在机制。方法: 选取肾移植术后 6 个月以上的受者作为研究对象，利用单细胞RNA测序（single-cell RNA sequencing，scRNA-seq）技术，对受者外周血单个核细胞（peripheral blood mononuclear cell，PBMC）中的免疫细胞亚群展开高分辨率的scRNA-seq分析。采用流式细胞技术检测不同免疫细胞群中差异基因对ABMR 的影响。结果: 本研究通过单细胞测序技术分析了ABMR受者与正常对照组的差异基因表达，发现CD69、CD83、CD52、CD74和CX3CR1为主要的差异基因。在经典单核细胞中，ABMR受者的CD83和CD52基因表达量较正常组有所增加，而治疗后呈现降低趋势；在初始CD4⁺T细胞中，ABMR受者的CD69基因表达量较正常组显著下降；在NK细胞中，ABMR受者的CX3CR1基因表达量较正常组降低，而治疗后CX3CR1的几何平均值进一步降低，CD74的几何平均值则有增加趋势。结论: 经典单核细胞中 CD83 和 CD52 基因表达上调，与抗原提呈细胞的功能增强相关，从而促进 ABMR 的发生。初始 CD4⁺T 细胞中 CD69 基因表达上调，与 T 细胞的激活和增殖相关，进一步参与ABMR 的进程。相反，NK 细胞中 CX3CR1 基因表达下调，与 NK 细胞的免疫监视功能障碍相关，从而在一定程度上抑制 ABMR 的发生。

关键词: 他克莫司, 肾移植, 排斥反应, 差异基因, PBMC

Abstract:

AIM: Based on single-cell sequencing analysis, the underlying mechanism of antibody-mediated rejection (ABMR) in kidney transplant recipients. METHODS: Recipients with 6 months or more after kidney transplantation were selected as research subjects, and high-resolution scRNA-seq analysis of immune cell subsets in peripheral blood mononuclear cells (PBMC) was performed using single-cell RNA sequencing (scRNA-seq) technology. Flow cytometry was used to examine the effect of differential genes on renal transplant ABMR in different immune cell populations. RESULTS: This study analyzed differential gene expression between ABMR patients and healthy controls using single-cell sequencing technology, identifying CD69, CD83, CD52, CD74, and CX3CR1 as key differentially expressed genes. In classical monocytes, ABMR patients exhibited increased CD83 and CD52 gene expression levels compared to the control group, which showed a downward trend after treatment. Regarding initial CD4⁺T cells, ABMR patients demonstrated significantly reduced CD69 gene expression. In NK cells, ABMR patients displayed decreased CX3CR1 gene expression initially, with further reduction in its geometric mean value post-treatment. Conversely, the geometric mean value of CD74 showed an increasing trend. CONCLUSION: In classical mononuclear cells, upregulated expression of CD83 and CD52 genes correlates with enhanced antigen-presenting cell functionality, thereby promoting the development of antigen-bonded microenvironment remodeling (ABMR). In initial CD4⁺T cells, elevated CD69 gene expression is associated with T cell activation and proliferation, further contributing to ABMR progression. Conversely, downregulated CX3CR1 gene expression in NK cells relates to impaired immune surveillance mechanisms, which partially inhibits ABMR formation.

Key words: tacrolimus, kidney transplantation, rejection, differential gene, PBMC

中图分类号:

郑凯乐, 付嘉钊, 尤佳, 吴丹, 王学彬, 王卓. 单细胞测序揭示肾移植受者抗体介导排斥反应的潜在机制[J]. 中国临床药理学与治疗学, 2026, 31(3): 289-299.

Kaile ZHENG, Jiazhao FU, Jia YOU, Dan WU, Xuebin WANG, Zhuo WANG. Single-cell sequencing reveals the underlying mechanism of antibody-mediated rejection in renal transplant recipients[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2026, 31(3): 289-299.

图/表 12

参考文献 30

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Cell type	Marker genes
T cell	CD3E, CD3D, CD3G
B cell	CD79B, CD19, CD79A
Fibroblast	COL1A1, COL1A2, COL3A1, POSTN
Epithelial cell	EPCAM, CDH1, KRT18, KRT8
Neutrophil	Ly6g, Ncf1, Csf3r, Cd177, Sorl1
Macrophage	CD14, CD68, CD163, CD209, AIF1
Dendritic cell	Flt3, CD1C
Natural killer cell	Nkg7, Ifng, Klrd1, Klrc1, CD56
Endothelial cell	VWF, CDH5, Pecam1
Basophil	TPSB2, TPSAB1, TPSAB2, CPA3
Plasma cell	CD27, CD38, XBP1, JCHAIN
Monocyte	CD14, CD300E, CD244
CD8⁺NKT-like cell	CD3E, CD8A, CD161, KLRC1
Classical monocyte	CD14, LYZ
Na?ve B cell	CD19, CD20, CD79A, IGLL1
Na?ve CD4⁺T cell	CD3E, CD4, CD45RA, LEF1
Na?ve CD8⁺T cell	CD3E, CD8A, CD45RA, LEF1
Non-classical monocyte	CD14, CD16, FCGR3A
Plasmacytoid dendritic cell	CD123, CD303, IRF7, LY6E
Platelet	CD41, CD61, PF4, GPIBα
Pro-B cell	CD19, CD34, CD79A, IGLL1, RAG1

Cell type	Marker genes
T cell	CD3E, CD3D, CD3G
B cell	CD79B, CD19, CD79A
Fibroblast	COL1A1, COL1A2, COL3A1, POSTN
Epithelial cell	EPCAM, CDH1, KRT18, KRT8
Neutrophil	Ly6g, Ncf1, Csf3r, Cd177, Sorl1
Macrophage	CD14, CD68, CD163, CD209, AIF1
Dendritic cell	Flt3, CD1C
Natural killer cell	Nkg7, Ifng, Klrd1, Klrc1, CD56
Endothelial cell	VWF, CDH5, Pecam1
Basophil	TPSB2, TPSAB1, TPSAB2, CPA3
Plasma cell	CD27, CD38, XBP1, JCHAIN
Monocyte	CD14, CD300E, CD244
CD8⁺NKT-like cell	CD3E, CD8A, CD161, KLRC1
Classical monocyte	CD14, LYZ
Na?ve B cell	CD19, CD20, CD79A, IGLL1
Na?ve CD4⁺T cell	CD3E, CD4, CD45RA, LEF1
Na?ve CD8⁺T cell	CD3E, CD8A, CD45RA, LEF1
Non-classical monocyte	CD14, CD16, FCGR3A
Plasmacytoid dendritic cell	CD123, CD303, IRF7, LY6E
Platelet	CD41, CD61, PF4, GPIBα
Pro-B cell	CD19, CD34, CD79A, IGLL1, RAG1

Variable	Recipient 1	Recipient 2	Recipient 3	Recipient 4
Gender	Male	Female	Male	Male
Age	15	48	54	51
Weight (kg)	41	58.1	68.5	85
Clinical diagnosis	AR	ABMR	IgAN	Normal
Organ transplantation	DCD	DCD	DCD	DCD
Immune induction	ATG	basiliximab	ATG	ATG
Immunosuppressant	TAC+MPA	TAC+MPA+Pred	TAC+MPA+Pred	TAC+MPA+Pred
Tacrolimus C₀ (ng/mL)	4.5	11.1	2.1	7.9
Treatment adjustment	Rituximab	Bortezomib	No adjustment	No adjustment
Serum creatinine (μmoI/L)	144	172	680	89
Uric acid (μmol/L)	593	345	303	498
Urea (mmol/L)	15.9	16.1	14.7	4.7
Albumin/globulin	139	1.18	1.19	1.54
Clinical manifestation	Normal body temperature, stable blood pressure	Normal body temperature, stable blood pressure	Normal body temperature, stable blood pressure	Normal body temperature, stable blood pressure

Variable	Recipient 1	Recipient 2	Recipient 3	Recipient 4
Gender	Male	Female	Male	Male
Age	15	48	54	51
Weight (kg)	41	58.1	68.5	85
Clinical diagnosis	AR	ABMR	IgAN	Normal
Organ transplantation	DCD	DCD	DCD	DCD
Immune induction	ATG	basiliximab	ATG	ATG
Immunosuppressant	TAC+MPA	TAC+MPA+Pred	TAC+MPA+Pred	TAC+MPA+Pred
Tacrolimus C₀ (ng/mL)	4.5	11.1	2.1	7.9
Treatment adjustment	Rituximab	Bortezomib	No adjustment	No adjustment
Serum creatinine (μmoI/L)	144	172	680	89
Uric acid (μmol/L)	593	345	303	498
Urea (mmol/L)	15.9	16.1	14.7	4.7
Albumin/globulin	139	1.18	1.19	1.54
Clinical manifestation	Normal body temperature, stable blood pressure	Normal body temperature, stable blood pressure	Normal body temperature, stable blood pressure	Normal body temperature, stable blood pressure

Renal transplant recipient samples	CD83 (PC7-A)	CD52 (APC)
ABMR_pre (03-L1-F1-1)	12173.0	5753.7
ABMR_post (06-L H 17-C1)	7377.2	3130.1
Normal (04-W1-F1)	10419.7	6564.7

单细胞测序揭示肾移植受者抗体介导排斥反应的潜在机制

Single-cell sequencing reveals the underlying mechanism of antibody-mediated rejection in renal transplant recipients

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参考文献 30

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Renal transplant recipient samples	CD69 (APC)
ABMR_pre (03-L1-F2-1)	2354.3
ABMR_post (06-L H 18-C2)	1416.5
Normal (04-W1-F2)	4922.9

Renal transplant recipient samples	CX3CR1 (PB450)	CD74 (APC)
ABMR_pre (03-L1-F3-1)	1471.8	472.5
ABMR_post (06-L H 19-C3)	936.1	513.9
Normal (07-GLB 19-C3)	1432.7	660.6

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