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| Cy7.SE标记ICOS-Ab荧光示踪技术评估小鼠心脏移植术后急性排斥反应 |
| 阳揭宇1△,傅宏1△,王全兴2,刘芳1,施晓敏1,郭闻渊1,宋少华1,李瑞东1,傅志仁1,丁国善1* |
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(1. 第二军医大学长征医院器官移植科,上海 200003;
2. 第二军医大学基础部免疫学教研室,上海 200433
△共同第一作者
*通信作者) |
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| 摘要: |
| 目的 探讨以T细胞活化表达的可诱导共刺激分子(inducible co-stimulatory molecule, ICOS)为基础的荧光示踪技术无创诊断小鼠心脏移植后急性排斥反应的可行性。方法 以小鼠颈部异位心脏移植为模型,分为同系移植、同种移植、同种移植+他克莫司(tacrolimus,FK506)治疗(同种移植治疗组)及同种移植治疗后停药组,分别在移植术后1、3、5、7 d,采用荧光染料Cy7.SE标记抗小鼠ICOS抗体(Cy7.SE-ICOS-Ab)经尾静脉注入移植小鼠,用荧光实时显像仪观察移植物荧光图像变化；同时用流式细胞仪检测各组小鼠脾脏T细胞表面ICOS表达,H-E染色分析心脏移植物病理改变。结果 同系移植及同种移植治疗组小鼠移植术后各时间点基本不显影；同种移植术后1 d,移植物荧光图像无明显变化,但术后3、5、7 d荧光逐渐增强,术后7 d时荧光强度最明显；同种移植治疗后停药组停药3 d移植物荧光强度逐渐增强,停药5、7 d后强度明显增强。H-E染色表明:各时间点同系移植及同种移植治疗组小鼠心脏移植物无明显排斥反应；同种移植及同种移植治疗后停药组在术后(或停药后)1 d移植物无明显排斥反应,术后3、5、7 d排斥反应逐渐出现并加重。流式细胞仪检测显示:术后1 d,各组脾脏T细胞表达ICOS水平无明显差异；同系移植及同种移植治疗组在各时间点基本不表达ICOS,但同种移植及同种移植治疗后停药组在术后(或停药后)3、5、7 d ICOS表达逐渐增加(P<0.05)。结论 ICOS表达强度与心脏移植后排斥反应强度有关,荧光标记的ICOS抗体可能有助于无创评估移植术后急性排斥反应的发生及程度。 |
| 关键词: 心脏移植 可诱导共刺激分子 移植物排斥 他克莫司 荧光抗体技术 |
| DOI：10.3724/SP.J.1008.2012.00721 |
| 投稿时间：2012-04-12修订日期：2012-06-06 |
| 基金项目:上海市科委医学重点项目(09411952300). |
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| Fluorescent tracer technique using ICOS-Ab marked with Cy7.SE for diagnosing acute heart graft rejection in mice |
| Fluorescent tracer technique using ICOS-Ab marked with Cy7.SE for diagnosing acute heart graft rejection in mice |
(1. Department of Organ Transplantation, Changzheng Hospital, Second Military Medical University, Shanghai 200003, China;
2. Department of Immunology, College of Basic Medical Sciences, Second Military Medical Univercity, Shanghai 200433, China
△Co-first authors.
*Corresponding author.) |
| Abstract: |
| ObjectiveTo establish a non-invasive method based on fluorescent tracer technique using inducible co-stimulatory molecules(ICOS) expressed on activated T cells for diagnosing acute heart graft rejection in mice. MethodsThe cervical heterotopic heart transplantation was used as model to establish isograft, allograft, allograft plus tacrolimus treatment, and allograft with tacrolimus ceased groups. On the 1st, 3rd, 5th and 7th day after transplantation, Cy7.SE-ICOS-Ab was injected into the heart transplant mice via tail veins. The real-time fluorescent imaging changes of the graft were observed by fluorescent equipment. Flow cytometry was used to examine the expression of ICOS on spleen T cells of mice in each group. H-E staining was used to observe the pathological changes of cardiac graft. ResultsThere was no noticeable fluorescent imaging in the grafts at the 1st, 3rd, 5th and 7th day after transplantation in the isograft and allograft with tacrolimus treatment group. On the first day after transplantation, the fluorescent imaging of graft in the allograft group had no noticeable changes, but the fluorescent imaging gradually increased on the 3rd, 5th, and 7th day. The graft fluorescent imaging became stronger on the 3rd day after ceasing tacrolimus in the treated allograft group, and it became stronger at 5 and 7 days after ceasing tacrolimus. H-E staining found no noticeable rejection in isograft group and allograft plus tacrolimus treatment group at all time points. The allograft and allograft puls tacrolimus ceased group developed rejection on the 3rd day after transplantation, and the rejection became more serious on the 5th and 7th day. Flow cytometry showed that there were no significant differences in ICOS expression on spleen T cells on the 1st day after transplantation among the four groups (P>0.05). The isograft and allograft plus tacrolimus treatment group had no ICOS expression on the T cells, and ICOS expression in the allograft and allograft with tacrolimus ceased group gradually increased on the 3rd, 5th and 7th day. ConclusionICOS expression intensity is associated with the degree of graft rejection. Fluorescently labeled anti-ICOS can help to assess the development and severity of acute rejection after transplantation in a non-invasive way. |
| Key words: heart transplantation inducible co-stimulatory molecules graft rejection tacrolimus fluorescent antibody technique |
