Robertson H, Wheeler J, Kirby J A, Morley A R
Department of Microbiology, The Medical School, University of Newcastle upon Tyne, United Kingdom.
Transplantation. 1996 May 27;61(10):1546-9. doi: 10.1097/00007890-199605270-00023.
A nonisotopic in situ hybridization method to detect perforin mRNA was developed in cytospin preparations of IL-2-stimulated normal human lymphocytes and applied to formalin-fixed acutely rejected renal transplant material. Individual cells expressing perforin mRNA were localized in severely damaged tubular areas, and a number of these cells appeared to be located inside the tubular basement membrane in close association with tubular epithelial cells. Immunoperoxidase staining in acetone-fixed cryostat sections of acutely rejected kidney confirmed that a considerable proportion of infiltrating cells was CD8+; many of these were in an intratubular location. In addition, perforin protein was identified in individual cells in similar locations to perforin mRNA-positive cells. Again, some intratubular cells were identified. Our findings illustrate that these cells can be fully activated with definite cytotoxic potential. Previously we have demonstrated that T lymphocytes proliferate within the tubular compartment during tubulitis, a characteristic condition in acute renal allograft rejection, and that there is associated tubular epithelial cell proliferation. In this study we think that we have further clarified the consequences of invasion of tubules by lymphoid cells. Our in situ hybridization method in rapid and convenient and may be applied to archival material.
我们开发了一种非同位素原位杂交方法，用于检测白细胞介素-2刺激的正常人淋巴细胞的细胞涂片制剂中的穿孔素mRNA，并将其应用于福尔马林固定的急性排斥肾移植材料。表达穿孔素mRNA的单个细胞定位于严重受损的肾小管区域，其中许多细胞似乎位于肾小管基底膜内，与肾小管上皮细胞紧密相连。急性排斥肾的丙酮固定冰冻切片的免疫过氧化物酶染色证实，相当一部分浸润细胞为CD8+；其中许多位于肾小管内。此外，在与穿孔素mRNA阳性细胞相似的位置的单个细胞中鉴定出了穿孔素蛋白。同样，也鉴定出了一些肾小管内细胞。我们的研究结果表明，这些细胞可以被充分激活并具有明确的细胞毒性潜力。此前我们已经证明，在肾小管炎（急性肾移植排斥反应的一种特征性情况）期间，T淋巴细胞在肾小管腔内增殖，并且伴有肾小管上皮细胞增殖。在本研究中，我们认为我们进一步阐明了淋巴细胞侵入肾小管的后果。我们的原位杂交方法快速便捷，可应用于存档材料。
