上海交通大学学报（医学版）

›› 2010, Vol. 30 ›› Issue (9): 1143-.doi: 10.3969/j.issn.1674-8115.2010.09.029

• 论著（临床研究） • 上一篇    下一篇

转移性体内迟发型超敏反应监测法的建立及临床应用

张明明, 周佩军, 邵 琨, 王祥慧, 徐 达   

  1. 上海交通大学 医学院附属瑞金医院泌尿外科肾移植中心, 上海 200025
  • 出版日期:2010-09-25 发布日期:2010-09-27
  • 通讯作者: 周佩军, 电子信箱: peijunzhou@yahoo.com。
  • 作者简介:张明明（1984—）, 男, 硕士生；电子信箱: zhangming2336@yahoo.com.cn。
  • 基金资助:

    上海市科委生物医药重点项目（09411952600）

Establishment of Trans-vivo delayed type hypersensitivity assay and its clinical application

ZHANG Ming-ming, ZHOU Pei-jun, SHAO Kun, WANG Xiang-hui, XU Da   

  1. Department of Urology, Renal Transplantation Center, Ruijin Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200025, China
  • Online:2010-09-25 Published:2010-09-27
  • Supported by:

    Biomedical Program of Shanghai Science and Technology Committee, 09411952600

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摘要：

目的 建立体内转移性迟发型超敏反应（Trans-vivo DTH）检测方法，用于评价临床移植受者的免疫状态。方法 通过Ficoll密度梯度离心法获取供者和受者（各13例）外周血单个核细胞（PBMC），用超声粉碎供者PBMC，制备供者亚细胞抗原。将受者的PBMC、回忆性抗原EBV抗原和供者抗原按以下各组分别注射入CB-17 SCID小鼠的足垫：①阴性对照组：7~9×106个受者PBMC；②供者抗原反应组：7~9×106个受者PBMC＋10 μg供者抗原；③阳性对照组：7~9×106个受者PBMC＋10 μg EBV抗原；④连锁免疫抑制组：7~9×106个受者PBMC＋10 μg EBV抗原＋10 μg供者抗原。分别于注射前及注射24 h后双盲测量小鼠的足垫厚度，计算两次测量厚度的差值。实验的有效性依据为阳性对照组足垫净厚度改变≥63.5×10-3 mm。结果 13例亲属活体肾移植受者接受Trans-vivo DTH检测显示，致敏型8例（62%），调节型3例（23%），非调节型2例（15%）。结论 Trans-vivo DTH检测法可用于监测移植后受者对供者抗原的反应性。

关键词: 免疫监测, 体内转移性迟发型超敏反应, 连锁免疫抑制, CB-17 SCID小鼠

Abstract:

Objective To establish trans-vivo delayed type hypersensitivity (trans-vivo DTH) assay for evaluation of immune response status in renal transplant recipients. Methods Peripheral blood mononuclear cells (PBMC) were obtained from donors (n=13) and recipients (n=13) by Ficoll density gradient centrifugation. Sub-cell antigens were prepared by ultrasonication of PBMC from donors. PBMC from recipients, recall antigen EBV antigens and donor antigens were injected into the foot pads of CB-17 SCID mice accordingly. For negative control group, 7-9×106 PBMC from recipients were injected. For donor antigen reactive group, 7-9×106 PBMC from recipients and 10 μg donor antigens were injected. For positive control group, 7-9 ×106 PBMC from recipients and 10 μg EBV antigens were injected. For linked immune suppression group, 7-9×106 PBMC from recipients, 10 μg EBV antigens and 10 μg donor antigens were injected. The thickness of foot pads of mice was measured before injection and 24 h after injection, and the change of thickness of foot pads was calculated. The net change of thickness ≥ 63.5×10-3 mm in positive control group was considered to be valid. Results Trans-vivo DTH assay in monitoring of 13 living related renal transplant recipients revealed that 8 patients (62%) were sensitized pattern, 3 patients (23%) were regulator pattern, and the other 2 patients (15%) were non-regulator pattern. Conclusion Response to donor antigens in living related renal transplant recipients can be monitored by trans-vivo DTH assay.

Key words: immunologic monitoring, Trans-vivo delayed type hypersensitivity, linked immune suppression, CB-17 SCID mouse