Laboratory of Immunology, Health Sciences Institute, Federal University of Bahia, Brazil.
Arch Oral Biol. 2012 Mar;57(3):314-20. doi: 10.1016/j.archoralbio.2011.09.003. Epub 2011 Sep 25.
Modulation of cell-mediated immunity by microorganisms in periodontal diseases has been widely studied; however, the proliferative activity and/or programmed death of mononuclear cells under periodontopathogenic stimuli are not yet well understood. The aim of this study was to investigate in vitro proliferation and death of peripheral blood mononuclear cells (PBMC) upon stimulation with Porphyromonas gingivalis (Pg) antigens.
In 19 patients with chronic periodontitis (CP) and 16 controls without periodontitis (NP) the following clinical parameters were evaluated: bleeding on probing, probing depth, and clinical attachment level. PBMC were cultured under Pg stimuli and apoptosis/necrosis and proliferation assays were carried out for 18 and 48 h, respectively. Fluorescence of labelled cells was determined using flow cytometry.
PBMC of CP and NP subjects exhibited a lower proliferative response to Pg LPS (p<0.05) and HmuY protein (p<0.001) compared with non-stimulated cells. Early apoptosis was induced by Pg LPS (p<0.01) and Pg extract (p<0.05), whilst all antigens induced late apoptosis (Pg LPS: p<0.001; Pg extract: p<0.001; HmuY: p<0.01) and necrosis (Pg LPS: p<0.01; Pg extract: p<0.001; HmuY: p<0.001). Pg LPS induced higher late apoptosis than HmuY (p<0.05). Only Pg LPS-induced necrosis tended to be higher in CP compared with NP.
The inhibitory effect of cell proliferation caused by Pg LPS and HmuY protein is not observed when these antigens comprise Pg extract. Despite induced apoptosis, some still unknown mechanism determines the inflammatory outcome in cell death stimulated by HmuY.
牙周病中微生物对细胞介导免疫的调节作用已得到广泛研究;然而,单核细胞在牙周致病菌刺激下的增殖活性和/或程序性死亡仍未得到很好的理解。本研究旨在研究体外刺激牙龈卟啉单胞菌(Pg)抗原对人外周血单个核细胞(PBMC)增殖和死亡的影响。
在 19 例慢性牙周炎(CP)患者和 16 例非牙周炎(NP)对照者中,评估以下临床参数:探诊出血、探诊深度和临床附着水平。PBMC 在 Pg 刺激下培养,并分别进行凋亡/坏死和增殖检测 18 和 48 小时。用流式细胞术测定标记细胞的荧光。
CP 和 NP 组 PBMC 对 Pg LPS(p<0.05)和 HmuY 蛋白(p<0.001)的增殖反应低于未刺激细胞。Pg LPS(p<0.01)和 Pg 提取物(p<0.05)诱导早期凋亡,而所有抗原均诱导晚期凋亡(Pg LPS:p<0.001;Pg 提取物:p<0.001;HmuY:p<0.01)和坏死(Pg LPS:p<0.01;Pg 提取物:p<0.001;HmuY:p<0.001)。Pg LPS 诱导的晚期凋亡高于 HmuY(p<0.05)。只有 Pg LPS 诱导的坏死在 CP 中比 NP 中更倾向于升高。
当这些抗原包含 Pg 提取物时,Pg LPS 和 HmuY 蛋白对细胞增殖的抑制作用并不明显。尽管诱导了凋亡,但在 HmuY 刺激的细胞死亡中,一些仍未知的机制决定了炎症结果。