Baqui A A, Meiller T F, Chon J J, Turng B F, Falkler W A
Department of Oral Medicine, Dental School, University of Maryland, Baltimore 21201, USA.
Clin Diagn Lab Immunol. 1998 May;5(3):341-7. doi: 10.1128/CDLI.5.3.341-347.1998.
Cytokines, including granulocyte-macrophage colony-stimulating factor (GM-CSF), are used to assist in bone marrow recovery during cancer chemotherapy. Interleukin-1beta (IL-1beta) and tumor necrosis factor alpha (TNF-alpha) play important roles in inflammatory processes, including exacerbation of periodontal diseases, one of the most common complications in patients who undergo this therapy. A human monocyte cell line (THP-1) was utilized to investigate IL-1beta and TNF-alpha production following GM-CSF supplementation with lipopolysaccharide (LPS) from two oral microorganisms, Porphyromonas gingivalis and Fusobacterium nucleatum. LPS of P. gingivalis or F. nucleatum was prepared by a phenol-water extraction method and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and determination of total protein and endotoxin contents. Resting THP-1 cells were treated with LPS of P. gingivalis or F. nucleatum and/or GM-CSF (50 IU/ml) by using different concentrations for various time periods. Production of IL-1beta and TNF-alpha in THP-1 cells was measured by solid-phase enzyme-linked immunosorbent assay. Reverse transcription (RT)-PCR was used to evaluate the gene expression of resting and treated THP-1 cells. IL-1beta was not detected in untreated THP-1 cells. IL-1beta production was, however, stimulated sharply at 4 h. GM-CSF amplified IL-1beta production in THP-1 cells treated with LPS from both oral anaerobes. No IL-1beta-specific mRNA transcript was detected in untreated THP-1 cells. However, IL-1beta mRNA was detected by RT-PCR 2 h after stimulation of THP-1 cells with LPS from both organisms. GM-CSF did not shorten the IL-1beta transcriptional activation time. GM-CSF plus F. nucleatum or P. gingivalis LPS activated THP-1 cells to produce a 1.6-fold increase in TNF-alpha production at 4 h over LPS stimulation alone. These investigations with the in vitro THP-1 model indicate that there may be an increase in the cellular immune response to oral endotoxin following GM-CSF therapy, as evidenced by production of the tissue-reactive cytokines IL-1beta and TNF-alpha.
细胞因子，包括粒细胞-巨噬细胞集落刺激因子（GM-CSF），被用于在癌症化疗期间协助骨髓恢复。白细胞介素-1β（IL-1β）和肿瘤坏死因子α（TNF-α）在炎症过程中发挥重要作用，包括牙周疾病的加重，牙周疾病是接受这种治疗的患者中最常见的并发症之一。利用人单核细胞系（THP-1）来研究在补充GM-CSF后，来自两种口腔微生物牙龈卟啉单胞菌和具核梭杆菌的脂多糖（LPS）刺激下IL-1β和TNF-α的产生情况。牙龈卟啉单胞菌或具核梭杆菌的LPS通过酚-水提取法制备，并通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及总蛋白和内毒素含量的测定进行表征。将静息的THP-1细胞用牙龈卟啉单胞菌或具核梭杆菌的LPS和/或GM-CSF（50 IU/ml）以不同浓度处理不同时间段。通过固相酶联免疫吸附测定法测量THP-1细胞中IL-1β和TNF-α的产生。逆转录（RT）-PCR用于评估静息和处理后的THP-1细胞的基因表达。在未处理的THP-1细胞中未检测到IL-1β。然而，在4小时时IL-1β的产生急剧增加。GM-CSF增强了用两种口腔厌氧菌的LPS处理的THP-1细胞中IL-1β的产生。在未处理的THP-1细胞中未检测到IL--1β特异性mRNA转录本。然而，在用两种微生物的LPS刺激THP-1细胞2小时后，通过RT-PCR检测到IL-1β mRNA。GM-CSF并未缩短IL-1β转录激活时间。GM-CSF加具核梭杆菌或牙龈卟啉单胞菌LPS在4小时时激活THP-1细胞，使其TNF-α产生量比单独的LPS刺激增加1.6倍。这些在体外THP-1模型中的研究表明，GM-CSF治疗后对口腔内毒素的细胞免疫反应可能会增加，组织反应性细胞因子IL-1β和TNF-α的产生证明了这一点。
