Madrenas J, Pazderka F, Parfrey N A, Halloran P F
Department of Medicine, University of Alberta, Edmonton, Canada.
J Immunol. 1992 Jan 15;148(2):612-9.
During studies on gene expression in the kidney, we unexpectedly observed that murine kidney expresses a truncated form of TCR-alpha mRNA (1.3-1.4 kb). This transcript was not associated with the presence of complete TCR-alpha mRNA (1.7 kb) or detectable TCR-beta or -delta transcripts, thus indicating that the truncated TCR-alpha mRNA could not be attributed to blood contamination of the kidney RNA preparation. The truncated TCR-alpha message appeared to contain at least the C alpha region, as suggested by hybridization with an intra-C alpha 24 oligonucleotide probe, by amplification of the C alpha region with the polymerase chain reaction from total kidney mRNA, and by sequencing of, and hybridization with, the amplified products. In situ hybridization of kidney sections indicated that the transcript was expressed in interstitial cells. Northern blots of cortex and medulla RNA showed that the cells expressing the truncated TCR-alpha mRNA were predominantly located in the medulla. To investigate the possibility that the transcript was not produced by T cells or NK cells, fractionation of renal cell suspensions were performed. The truncated TCR-alpha mRNA was detected in a fraction containing large (low buoyant density) cells in which no expression of CD3, Thy 1, or NK-1.1 was detected, indicating that these cells are not mature T cells, do not express a functional TCR, and are not NK cells. The cells expressing the truncated TCR-alpha mRNA were radiosensitive, and were not thymus dependent, because this transcript was as abundant in nude mice as in normal mice. The transcript was not detected in bone marrow. Expression of the truncated TCR-alpha mRNA was not dependent on an intact recombinase activity as its expression was not affected by the severe combined immunodeficiency mutation. Our results show that murine kidney contains a population of radiosensitive thymus-independent large interstitial cells that express a truncated TCR-alpha mRNA that is not associated with surface expression of functional TCR. These cells may have attempted to rearrange TCR-alpha genes, suggesting that they may be related to the lymphoid lineage.
在对肾脏基因表达的研究过程中,我们意外地观察到小鼠肾脏表达一种截短形式的TCR-α mRNA(1.3 - 1.4 kb)。该转录本与完整的TCR-α mRNA(1.7 kb)的存在或可检测到的TCR-β或 -δ转录本无关,因此表明截短的TCR-α mRNA并非归因于肾脏RNA制备过程中的血液污染。截短的TCR-α信息似乎至少包含Cα区域,这通过与Cα内部24寡核苷酸探针杂交、从总肾脏mRNA中通过聚合酶链反应扩增Cα区域以及对扩增产物进行测序和杂交得以表明。肾脏切片的原位杂交表明该转录本在间质细胞中表达。皮质和髓质RNA的Northern印迹显示,表达截短的TCR-α mRNA的细胞主要位于髓质。为了研究该转录本并非由T细胞或NK细胞产生的可能性,我们对肾细胞悬液进行了分离。在含有大(低浮力密度)细胞的部分中检测到了截短的TCR-α mRNA,在这些细胞中未检测到CD3、Thy 1或NK-1.1的表达,这表明这些细胞不是成熟的T细胞,不表达功能性TCR,也不是NK细胞。表达截短的TCR-α mRNA的细胞对辐射敏感,且不依赖胸腺,因为该转录本在裸鼠中与正常小鼠中一样丰富。在骨髓中未检测到该转录本。截短的TCR-α mRNA的表达不依赖于完整的重组酶活性,因为其表达不受严重联合免疫缺陷突变的影响。我们的结果表明,小鼠肾脏含有一群对辐射敏感、不依赖胸腺的大间质细胞,它们表达一种截短的TCR-α mRNA,且与功能性TCR的表面表达无关。这些细胞可能尝试重排TCR-α基因,这表明它们可能与淋巴谱系相关。