Suzuki J, Isobe M, Yamazaki S, Horie S, Okubo Y, Sekiguchi M
First Department of Internal Medicine, School of Medicine, Shinshu University, Matsumoto, Japan.
Cardiovasc Res. 1998 Nov;40(2):307-13. doi: 10.1016/s0008-6363(98)00126-6.
In situ transcription of cytokines which are important in the development of cardiac rejection has not been evaluated for diagnosing rejection. The objective was to evaluate the usefulness of in situ reverse transcriptase polymerase chain reaction (RT-PCR) for sensitive detection of acute cardiac rejection.
We studied interferon (IFN)-gamma and interleukin (IL)-2 expression using immunohistochemistry and in situ RT-PCR in murine cardiac transplant models. Hearts were heterotopically transplanted (BALB/c to C3H/He) and some mice were not treated (n = 23); others were treated with anti-intercellular adhesion molecule (ICAM)-1 and anti-lymphocyte function associated antigen (LFA)-1 monoclonal antibodies (mAbs) (n = 23). Allografts were removed at days 1 to 7. For control, isografts were harvested at day 7 (n = 2).
Mice without treatment rejected allografts within 7 days, while all mAb-treated recipients accepted allografts for the same period. At day 1, allografts of both groups showed scattered myocardial cell infiltration which increased in non-treated allografts, but remained stable in mAb-treated grafts thereafter. In situ RT-PCR showed that IL-2 and IFN-gamma mRNA positive cells were present in non-treated allografts, while few mRNA positive cells were expressed in infiltrating cells in the mAb-treated allografts (IL-2, day 3: 88.8 +/- 28.3 vs. 7.2 +/- 6.4, p < 0.05, positive cells within 10 fields per section). However, immunohistochemistry could not reveal the difference at day 3.
In situ RT-PCR is a sensitive method for diagnosing acute rejection, and it reveals the characteristics of myocardial infiltrate cells to determine their role in the process of rejection.
在心脏排斥反应发生过程中起重要作用的细胞因子的原位转录尚未被评估用于诊断排斥反应。本研究的目的是评估原位逆转录聚合酶链反应（RT-PCR）用于敏感检测急性心脏排斥反应的实用性。
我们在小鼠心脏移植模型中使用免疫组织化学和原位RT-PCR研究了干扰素（IFN）-γ和白细胞介素（IL）-2的表达。心脏进行异位移植（从BALB/c到C3H/He），一些小鼠未接受治疗（n = 23）；其他小鼠用抗细胞间黏附分子（ICAM）-1和抗淋巴细胞功能相关抗原（LFA）-1单克隆抗体（mAb）治疗（n = 23）。在第1至7天切除同种异体移植物。作为对照，在第7天收获同基因移植物（n = 2）。
未治疗的小鼠在7天内排斥同种异体移植物，而所有接受mAb治疗的受体在同一时期接受了同种异体移植物。在第1天，两组的同种异体移植物均显示散在的心肌细胞浸润，未治疗的同种异体移植物中浸润增加，但此后接受mAb治疗的移植物中浸润保持稳定。原位RT-PCR显示，未治疗的同种异体移植物中存在IL-2和IFN-γ mRNA阳性细胞，而在接受mAb治疗的同种异体移植物的浸润细胞中很少有mRNA阳性细胞表达（IL-2，第3天：88.8 +/- 28.3对7.2 +/- 6.4，p < 0.05，每切片10个视野内的阳性细胞）。然而，免疫组织化学在第3天未能显示出差异。
原位RT-PCR是诊断急性排斥反应的一种敏感方法，它揭示了心肌浸润细胞的特征，以确定它们在排斥反应过程中的作用。
