Center for Oral Health Research, College of Dentistry, University of Kentucky, Lexington, KY 40536-0305, USA.
Mol Oral Microbiol. 2010 Apr;25(2):136-49. doi: 10.1111/j.2041-1014.2009.00552.x.
Bacterial and host cell products during coinfections of Human Immunodeficiency Virus type 1-positive (HIV-1(+)) patients regulate HIV-1 recrudescence in latently infected cells (e.g. T cells, monocytes/macrophages), impacting highly active antiretroviral therapy (HAART) failure and progression of acquired immunodeficiency syndrome. A high frequency of oral opportunistic infections (e.g. periodontitis) in HIV-1(+) patients has been demonstrated; however, their potential to impact HIV-1 exacerbation is unclear. We sought to determine the ability of supernatants derived from oral epithelial cells (OKF4) and human gingival fibroblasts (Gin-4) challenged with periodontal pathogens, to modulate the HIV-1 promoter activation in monocytes/macrophages. BF24 monocytes/macrophages transfected with the HIV-1 promoter driving the expression of chloramphenicol acetyltransferase (CAT) were stimulated with Porphyromonas gingivalis, Fusobacterium nucleatum, or Treponema denticola in the presence of supernatants from OKF4 or Gin4 cells either unstimulated or previously pulsed with bacteria. CAT levels were determined by enzyme-linked immunosorbent assay and cytokine production was evaluated by Luminex beadlyte assays. OKF4 and Gin4 supernatants enhanced HIV-1 promoter activation particularly related to F. nucleatum challenge. An additive effect was observed in HIV-1 promoter activation when monocytes/macrophages were simultaneously stimulated with gingival cell supernatants and bacterial extracts. OKF4 cells produced higher levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukins -6 and -8 in response to F. nucleatum and P. gingivalis. Preincubation of OKF4 supernatants with anti-GM-CSF reduced the additive effect in periodontopathogen-induced HIV-1 promoter activation. These results suggest that soluble mediators produced by gingival resident cells in response to periodontopathogens could contribute to HIV-1 promoter activation in monocytes/macrophages, albeit this effect is most notable following direct stimulation of the cells with oral gram-negative bacteria.
细菌和宿主细胞产物在 HIV-1 阳性(HIV-1(+))患者的合并感染中调节潜伏感染细胞(如 T 细胞、单核细胞/巨噬细胞)中的 HIV-1 复发,影响高效抗逆转录病毒治疗(HAART)失败和获得性免疫缺陷综合征的进展。已经证明 HIV-1(+)患者中口腔机会性感染(如牙周炎)的频率很高;然而,它们对 HIV-1 恶化的潜在影响尚不清楚。我们试图确定受牙周病病原体挑战的口腔上皮细胞(OKF4)和人牙龈成纤维细胞(Gin-4)产生的上清液调节单核细胞/巨噬细胞中 HIV-1 启动子激活的能力。BF24 单核细胞/巨噬细胞转染了 HIV-1 启动子,该启动子驱动氯霉素乙酰转移酶(CAT)的表达,在用 OKF4 或 Gin4 细胞上清液刺激的同时,用牙龈细胞或以前用细菌脉冲处理的 Porphyromonas gingivalis、Fusobacterium nucleatum 或 Treponema denticola 刺激,通过酶联免疫吸附测定法测定 CAT 水平,并通过 Luminex beadlyte 测定法评估细胞因子的产生。OKF4 和 Gin4 上清液增强了 HIV-1 启动子的激活,特别是与 F. nucleatum 挑战有关。当单核细胞/巨噬细胞同时受到牙龈细胞上清液和细菌提取物的刺激时,观察到 HIV-1 启动子激活的附加效应。OKF4 细胞在响应 F. nucleatum 和 P. gingivalis 时产生更高水平的粒细胞-巨噬细胞集落刺激因子(GM-CSF)和白细胞介素-6 和 -8。用抗 GM-CSF 预处理 OKF4 上清液可降低牙周病病原体诱导的 HIV-1 启动子激活中的附加效应。这些结果表明,牙龈常驻细胞对牙周病病原体的反应产生的可溶性介质可能有助于单核细胞/巨噬细胞中 HIV-1 启动子的激活,尽管这种作用在细胞直接受到口腔革兰氏阴性细菌刺激时最为明显。