Azad N, Emanuele N V, Halloran M M, Tentler J, Kelley M R
Research Service, Hines VA Hospital, IL 60141.
Endocrinology. 1991 Mar;128(3):1679-81. doi: 10.1210/endo-128-3-1679.
Pursuant to our report of an immunoreactive and bioactive luteinizing hormone-releasing hormone (LHRH)-like molecule in rat spleen lymphocytes, we sought to determine whether these cells were capable of synthesizing LHRH by determining whether lymphocytes contain LHRH mRNA. To do this, total RNA was extracted from hypothalamic tissue, anterior pituitaries and from lymphocytes, and then this was reverse transcribed to cDNA and amplified via the polymerase chain reaction (PCR) utilizing synthetic oligonucleotides bracketing a portion of the LHRH gene. Following gel electrophoresis a discrete band of the expected size of 375 base pairs was found in the hypothalamus (positive control), and in lymphocytes, but not in the anterior pituitary (negative control). Furthermore, after Southern blotting, a 32P-labelled LHRH cDNA, hybridized to the 375 base pair, was amplified in hypothalamus fragments and in lymphocytes, but not in anterior pituitary tissue. These data strongly suggest that LHRH, in addition to being an important neuropeptide, is an immune cell synthesized immunomodulator.
根据我们关于大鼠脾脏淋巴细胞中一种具有免疫反应性和生物活性的促黄体生成素释放激素（LHRH）样分子的报告，我们试图通过确定淋巴细胞是否含有LHRH mRNA来确定这些细胞是否能够合成LHRH。为此，从下丘脑组织、垂体前叶和淋巴细胞中提取总RNA，然后将其逆转录为cDNA，并使用围绕LHRH基因一部分的合成寡核苷酸通过聚合酶链反应（PCR）进行扩增。凝胶电泳后，在下丘脑（阳性对照）和淋巴细胞中发现了预期大小为375个碱基对的离散条带，但在垂体前叶（阴性对照）中未发现。此外，Southern印迹后，在375个碱基对处与32P标记的LHRH cDNA杂交的片段，在下丘脑片段和淋巴细胞中被扩增，但在垂体前叶组织中未被扩增。这些数据有力地表明，LHRH除了是一种重要的神经肽外，还是一种免疫细胞合成的免疫调节剂。
