Molina Moisés Armides Franco, Gamboa Edgar Mendoza, Tello Paloma Castillo, Benavides Pablo Zapata, León Leonardo Castillo, Guerra Reyes Tamez, Padilla Cristina Rodríguez
Laboratorio de Inmunología y Virología, Departamento de Microbiología e Inmunología. Facultad de Ciencias Biológicas, Universidad Autónoma de Nuevo León, Nuevo León, México.
Lymphat Res Biol. 2006 Spring;4(1):34-40. doi: 10.1089/lrb.2006.4.34.
Cytokines regulate cellular immune activity and are produced by a variety of cells, especially lymphocytes, monocytes, and macrophages. Measurement of cytokine levels has yielded useful information on the pathological process of different diseases such as AIDS, endotoxic shock, sepsis, asthma, and cancer. It may also be of use in the monitoring of disease progression and/or inflammation. To determine spontaneous cytokine gene expression in whole blood and PBMCs, whole blood was obtained from healthy volunteers and total mRNA was isolated from PBMCs. The kinetics of response were determined by sequential testing of cytokine gene expression by RT-PCR analysis. Our results demonstrated that isolated and incubated PBMCs expressed TNF-alpha and high levels of IL-1beta, IL-6, IL-8, and IL-10. In contrast, WB only expressed the mRNA cytokines of TNF-alpha and IL-8 (p < 0.05). These results suggest that spontaneous myriad mRNA cytokine expression can be avoided with the use of WB incubation and the rapid collection of PBMCs. Furthermore, this method should be employed in all cases where the levels of cytokine gene expression can be evaluated.
细胞因子调节细胞免疫活性，由多种细胞产生，尤其是淋巴细胞、单核细胞和巨噬细胞。细胞因子水平的测量为艾滋病、内毒素休克、败血症、哮喘和癌症等不同疾病的病理过程提供了有用信息。它也可能用于监测疾病进展和/或炎症。为了确定全血和外周血单个核细胞（PBMCs）中细胞因子基因的自发表达，从健康志愿者获取全血，并从PBMCs中分离总mRNA。通过逆转录聚合酶链反应（RT-PCR）分析对细胞因子基因表达进行连续检测，从而确定反应动力学。我们的结果表明，分离并孵育的PBMCs表达肿瘤坏死因子-α（TNF-α）以及高水平的白细胞介素-1β（IL-1β）、白细胞介素-6（IL-6）、白细胞介素-8（IL-8）和白细胞介素-10（IL-10）。相比之下，全血仅表达TNF-α和IL-8的mRNA细胞因子（p < 0.05）。这些结果表明，通过全血孵育和快速收集PBMCs可以避免自发的多种mRNA细胞因子表达。此外，在所有可以评估细胞因子基因表达水平的情况下都应采用此方法。
