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首页> 外文期刊>Cytokine >Quantification of human interleukin 18 mRNA expression by competitive reverse transcriptase polymerase chain reaction.
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Quantification of human interleukin 18 mRNA expression by competitive reverse transcriptase polymerase chain reaction.

机译：通过竞争性逆转录聚合酶链反应定量人类白细胞介素18 mRNA的表达。

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Interleukin 18 (IL-18) is a recently identified cytokine, originally called interferon gamma inducing factor, due to its capacity to induce interferon gamma production in Th1 type cells. IL-18 is expressed by a wide variety of cell types including mononuclear phagocytes, osteoblasts, keratinocytes and adrenal cortex cells. To quantify human IL-18 mRNA expression in small-scale cell samples the authors developed a competitive reverse transcriptase polymerase chain reaction using a competitive template as an internal standard. This assay was demonstrated as a valid, sensitive and precise tool to quantify human IL-18 mRNA expression. IL-18 mRNA expression of primary peripheral blood monocytes, CD4(+)T cells, CD8(+)T cells, B cells and NK cells was assessed by competitive RT-PCR. Basal IL-18 expression could be detected in all types of peripheral blood mononuclear cells (PBMC). The kinetics of IL-18 mRNA expression in PBMC from healthy donors was defined in vitro after monocyte-specific (lipopolysaccharide LPS), T-cell-specific (anti-CD3) and polyclonal-unspecific stimulation (phytohaemagglutinin PHA). Only LPS led to a strong increase of IL-18 mRNA expression peaking after 2 h. These results indicate that IL-18 is expressed constitutionally by all major PBMC subtypes. However, only monocyte specific stimulation resulted in a significant induction of IL-18 mRNA expression suggesting activated monocytes e.g. in inflammation as the main source of IL-18 expression. Copyright 1999 Academic Press.
机译：白介素18（IL-18）是最近被鉴定出的细胞因子，最初被称为干扰素γ诱导因子，因为它具有在Th1型细胞中诱导干扰素γ产生的能力。 IL-18由多种细胞类型表达，包括单核吞噬细胞，成骨细胞，角质形成细胞和肾上腺皮质细胞。为了量化小规模细胞样品中人IL-18 mRNA的表达，作者开发了一种竞争性逆转录聚合酶链反应，使用竞争性模板作为内标。该测定法被证明是定量人IL-18 mRNA表达的有效，灵敏和精确的工具。通过竞争性RT-PCR评估初级外周血单核细胞，CD4（+）T细胞，CD8（+）T细胞，B细胞和NK细胞的IL-18 mRNA表达。在所有类型的外周血单核细胞（PBMC）中均可检测到基础IL-18表达。在单核细胞特异性（脂多糖LPS），T细胞特异性（抗CD3）和多克隆非特异性刺激（植物血凝素PHA）刺激后，体外确定健康供体PBMC中IL-18 mRNA表达的动力学。仅LPS导致2小时后达到峰值的IL-18 mRNA表达强烈增加。这些结果表明，IL-18在所有主要的PBMC亚型中均表达。然而，仅单核细胞特异性刺激导致IL-18 mRNA表达的显着诱导，表明活化的单核细胞，例如单核细胞。在炎症中作为IL-18表达的主要来源。版权所有1999，学术出版社。

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