Lai J P, Douglas S D, Ho W Z
Joseph Stokes Jr. Research Institute at the Children's Hospital of Philadelphia, Department of Pediatrics, University of Pennsylvania Medical School, 19104, USA.
J Neuroimmunol. 1998 Jun 1;86(1):80-6. doi: 10.1016/s0165-5728(98)00025-3.
We present data demonstrating the gene expression of substance P (SP) and its receptor in human peripheral blood-isolated lymphocytes. Using reverse transcribed polymerase chain reaction (RT-PCR) assay, preprotachykinin-A (substance-P) mRNA is detected in human peripheral blood-isolated lymphocytes. Among the alpha, beta, and gamma transcripts of the SP gene, only the beta and gamma transcripts are detectable in these cells. These RT-PCR amplified transcripts are recognized by Southern blot assay using a specific SP probe. Direct DNA sequence analysis of the RT-PCR products from lymphocytes also confirmed the structure of these transcripts which are identical to those found in human neuronal cells. At the protein level, human lymphocytes produced endogenous SP as determined by an enzyme immunoassay. Capsaicin, a vanillyl fatty acid amide (ingredient of hot pepper), released preformed SP from lymphocytes. In addition, using RT/nested-PCR analysis, we identified the presence of mRNA for neurokinin-1 receptor (the receptor for SP) in human peripheral blood-isolated lymphocytes, which was confirmed by Southern blot and DNA sequencing analysis. The demonstration that human lymphocytes express SP and its receptor support the notion that SP is biologically involved in regulating the functions of these cells in an autocrine fashion.
我们提供的数据表明，P物质（SP）及其受体在人外周血分离淋巴细胞中存在基因表达。通过逆转录聚合酶链反应（RT-PCR）检测，在人外周血分离淋巴细胞中可检测到前速激肽原A（P物质）mRNA。在SP基因的α、β和γ转录本中，这些细胞中仅可检测到β和γ转录本。使用特异性SP探针的Southern印迹分析可识别这些RT-PCR扩增的转录本。对淋巴细胞RT-PCR产物的直接DNA序列分析也证实了这些转录本的结构，其与在人神经细胞中发现的结构相同。在蛋白质水平上，通过酶免疫测定法确定人淋巴细胞产生内源性SP。辣椒素，一种香草基脂肪酸酰胺（辣椒成分），可从淋巴细胞中释放预先形成的SP。此外，通过RT/巢式PCR分析，我们在人外周血分离淋巴细胞中鉴定出神经激肽-1受体（SP的受体）mRNA的存在，这通过Southern印迹和DNA序列分析得到证实。人淋巴细胞表达SP及其受体的证明支持了SP以自分泌方式生物学参与调节这些细胞功能的观点。
