Azad N, La Paglia N, Jurgens K A, Kirsteins L, Emanuele N V, Kelley M R, Lawrence A M, Mohagheghpour N
Department of Research, Department of Veterans Affairs, Edward Hines Jr. Hospital, Hines, Illinois 60141.
Endocrinology. 1993 Jul;133(1):215-23. doi: 10.1210/endo.133.1.8319570.
An immunomodulatory role for LHRH was suggested when we reported the presence of immunoactive and bioactive LHRH and its mRNA in rat splenic and thymic lymphocytes. In this paper we report that human peripheral T-cells as well as its subsets CD4+ and CD8+ contained immunoactive and bioactive LHRH. Furthermore, analysis of phytohemagglutinin (PHA)-activated T-cell lysates for LHRH by RIA demonstrated that the mean concentration of LHRH in PHA-activated T-cells increased from 45 +/- 4.5 to 64 +/- 7 pg/10(6) cells after 24 h of culture and from 47 +/- 3.6 to 117 +/- 11.8 pg/10(6) cells (P < 0.01) after 48 h. While the LHRH concentration in PHA-activated cells increased over the last 24 h of culture h from 64 +/- 7 to 117 +/- 11.8 pg/10(6) cells (P < 0.001), there was no change in mean concentration of LHRH in T-cells kept in medium alone. In a preliminary study we found that fresh T-cells contain 20 +/- 1.4 pg pro-LHRH/10(6) cells, and PHA stimulation increased the pro-LHRH content similar to the increase in LHRH. As with unfractionated T-cells, a significant PHA-induced time-dependent enhancement of intracellular LHRH was noted in CD4+ and CD8+ T-cells. RNA extracted from lymphocytes was subjected to reverse transcription-polymerase chain reaction analysis using LHRH and histone-3.3, primers, the latter as an internal control. The polymerase chain reaction-generated data demonstrated that the relative amount of LHRH mRNA in cultured, but non-PHA-stimulated (resting), cells diminished dramatically between 5-24 h, but recovered by 48 h of culture. The relative amount of LHRH mRNA in PHA-stimulated cells revealed a markedly different pattern. LHRH message expression in PHA-activated cells increased slightly at 5 h of culture and was maximally stimulated by 24 h, but declined by 48 h of culture. The PHA activation-induced time-dependent enhancement of intracellular accumulation of LHRH peptide at 5 and 24 h was accompanied by increased LHRH message. However, the increased concentration of LHRH peptide at 48 h coincided with decreased LHRH message expression. The data from total protein synthesis in PHA-activated cells showed a progressive increase in protein synthesis, a pattern entirely similar to the changes in the cell content of LHRH.(ABSTRACT TRUNCATED AT 400 WORDS)
当我们报道大鼠脾脏和胸腺淋巴细胞中存在免疫活性和生物活性促性腺激素释放激素（LHRH）及其信使核糖核酸（mRNA）时，提示LHRH具有免疫调节作用。在本文中，我们报道人类外周血T细胞及其CD4⁺和CD8⁺亚群含有免疫活性和生物活性LHRH。此外，通过放射免疫分析（RIA）对植物血凝素（PHA）激活的T细胞裂解物进行LHRH分析表明，培养24小时后，PHA激活的T细胞中LHRH的平均浓度从45±4.5增加到64±7皮克/10⁶个细胞，培养48小时后从47±3.6增加到117±11.8皮克/10⁶个细胞（P<0.01）。在培养的最后24小时内，PHA激活细胞中的LHRH浓度从64±7增加到117±11.8皮克/10⁶个细胞（P<0.001），而单独置于培养基中的T细胞中LHRH的平均浓度没有变化。在一项初步研究中，我们发现新鲜T细胞含有20±1.4皮克前体LHRH/10⁶个细胞，PHA刺激使前体LHRH含量增加，与LHRH的增加相似。与未分离的T细胞一样，在CD4⁺和CD8⁺T细胞中也观察到PHA诱导的细胞内LHRH随时间的显著增强。从淋巴细胞中提取的RNA使用LHRH和组蛋白-3.3引物进行逆转录-聚合酶链反应分析，后者作为内对照。聚合酶链反应产生的数据表明，培养但未受PHA刺激（静止）的细胞中LHRH mRNA的相对量在5至24小时之间急剧减少，但在培养48小时时恢复。PHA刺激细胞中LHRH mRNA的相对量呈现出明显不同的模式。PHA激活细胞中LHRH信息表达在培养5小时时略有增加，在24小时时受到最大刺激，但在培养48小时时下降。PHA激活诱导的LHRH肽在5小时和24小时细胞内积累随时间的增强伴随着LHRH信息的增加。然而，48小时时LHRH肽浓度的增加与LHRH信息表达的减少同时出现。PHA激活细胞中总蛋白质合成的数据显示蛋白质合成逐渐增加，这一模式与LHRH细胞含量的变化完全相似。（摘要截短至400字）
