Pachner A R, Amemiya K, Delaney E, O'Neill T, Hughes C A, Zhang W F
Department of Neurology, Georgetown University School of Medicine, Washington, DC 20007, USA.
Neurology. 1997 Jul;49(1):147-52. doi: 10.1212/wnl.49.1.147.
In patients with Lyme neuroborreliosis, inflammation and symptoms of fatigue and malaise occur out of proportion to the relatively low number of spirochetes present. Previous studies have identified interleukin-6 (IL-6) as a candidate molecule for amplification of CNS inflammation in this disease. We pursued this possibility by measuring cytokine gene expression by reverse-transcriptase polymerase chain reaction (RT-PCR) in the brain of rhesus macaques actively infected with Borrelia burgdorferi. Samples of brain tissue were screened for IL-6 and interferon gamma using RT-PCR-ELISA, a technique that uses RT-PCR, subsequent hybridization of the PCR product with a biotinylated probe, and capture and ELISA readout of hybridization product. The number of copies in positive samples was then quantitated using qRT-PCR-ELISA, in which wild-type cytokine cDNA competes with recombinant competitor DNA in the PCR. Elevated levels of IL-6 cDNA and, to a lesser extent, interferon gamma were detected in three of three nonhuman primates with persistent infection with B burgdorferi, whereas the brains of three uninfected animals and undetectable levels of gene expression of these cytokines. These data support the hypothesis that cytokines such as IL-6 are important amplification molecules for CNS inflammation in Lyme neuroborreliosis.
在莱姆病神经疏螺旋体病患者中，炎症以及疲劳和不适症状的出现与相对少量存在的螺旋体不成比例。先前的研究已将白细胞介素-6（IL-6）确定为该疾病中中枢神经系统炎症放大的候选分子。我们通过逆转录聚合酶链反应（RT-PCR）测量恒河猴大脑中细胞因子基因的表达来探究这种可能性，这些恒河猴被伯氏疏螺旋体主动感染。使用RT-PCR-ELISA对脑组织样本进行IL-6和干扰素γ筛查，该技术采用RT-PCR，随后将PCR产物与生物素化探针杂交，并对杂交产物进行捕获和ELISA读数。然后使用qRT-PCR-ELISA对阳性样本中的拷贝数进行定量，其中野生型细胞因子cDNA在PCR中与重组竞争DNA竞争。在三只持续感染伯氏疏螺旋体的非人灵长类动物中，检测到三只动物的IL-6 cDNA水平升高，干扰素γ水平在较小程度上升高，而三只未感染动物的大脑中这些细胞因子的基因表达水平检测不到。这些数据支持这样的假设，即诸如IL-6之类的细胞因子是莱姆病神经疏螺旋体病中枢神经系统炎症的重要放大分子。
