Institute of Oral Biology, National Yang-Ming University, Taipei, Taiwan.
J Periodontal Res. 2012 Oct;47(5):635-44. doi: 10.1111/j.1600-0765.2012.01476.x. Epub 2012 Apr 4.
Herpesviruses may play roles in the development of periodontal diseases. This study analyzed the effects of herpes simplex virus type 1 (HSV-1) infection on neutrophil function. The effects of lipopolysaccharide (LPS) from the periodontal pathogen, Porphyromonas gingivalis, during HSV-1 infection were also determined.
Purified HSV-1 was pretreated with buffer containing no serum, with HSV-1 immunoglobulin G (IgG)-positive serum (HSV-1 antiserum) or with control serum. Neutrophils were mock-infected or infected with the pretreated HSV-1. Viral binding and phagosome formation were detected using immunostaining. Intracellular reactive oxygen species (ROS) were determined using 2',7'-dichlorofluorescin diacetate and fluorometry. Leukotriene B(4) (LTB(4)) and interleukin-8 (IL-8) were detected using enzyme immunoassays. Release of matrix metalloproteinase-9 (MMP-9) was examined using gelatin zymography. Phosphorylation of Akt/glycogen synthase kinase-3 (GSK-3) was determined using western blotting.
HSV-1 bound directly to neutrophils and enhanced the release of MMP-9. HSV-1 immune complexes, formed in the HSV-1 antiserum, bound neutrophils and induced the formation of early phagosome more effectively than did HSV-1 alone. The relative levels of ROS and phosphorylation of Akt/GSK-3 were increased significantly in neutrophils after infection with HSV-1 immune complexes. Infection with HSV-1 and HSV-1 immune complexes also stimulated the production of inflammatory mediators, LTB(4) and IL-8. Moreover, LPS enhanced the HSV-1-stimulatory production of IL-8.
This study demonstrated differences in neutrophils infected with HSV-1 alone or with HSV-1 immune complexes, suggesting that opsonization of HSV-1 might enhance its effects on neutrophils. The in vitro findings suggest that HSV-1 infection may induce the inflammatory response and affect periodontal health.
疱疹病毒可能在牙周病的发展中起作用。本研究分析了单纯疱疹病毒 1 型（HSV-1）感染对中性粒细胞功能的影响。还确定了牙周病原体牙龈卟啉单胞菌的脂多糖（LPS）在 HSV-1 感染期间的作用。
用不含血清的缓冲液预处理纯化的 HSV-1，用 HSV-1 免疫球蛋白 G（IgG）阳性血清（HSV-1 抗血清）或对照血清预处理。用预处理的 HSV-1 模拟感染或感染中性粒细胞。用免疫染色检测病毒结合和吞噬体形成。用 2'，7'-二氯荧光素二乙酸酯和荧光法测定细胞内活性氧（ROS）。用酶联免疫吸附法检测白三烯 B(4)（LTB(4)）和白细胞介素-8（IL-8）。用明胶酶谱法检测基质金属蛋白酶-9（MMP-9）的释放。用 Western 印迹法测定 Akt/糖原合酶激酶-3（GSK-3）的磷酸化。
HSV-1 直接与中性粒细胞结合并增强 MMP-9 的释放。在 HSV-1 抗血清中形成的 HSV-1 免疫复合物比单独的 HSV-1 更有效地结合中性粒细胞并诱导早期吞噬体形成。感染 HSV-1 免疫复合物后，中性粒细胞中 ROS 和 Akt/GSK-3 的磷酸化水平显著升高。感染 HSV-1 和 HSV-1 免疫复合物也刺激炎症介质 LTB(4)和 IL-8 的产生。此外，LPS 增强了 HSV-1 刺激的 IL-8 产生。
本研究表明单独感染 HSV-1 或 HSV-1 免疫复合物的中性粒细胞之间存在差异，提示 HSV-1 的调理作用可能增强其对中性粒细胞的作用。体外研究结果表明，HSV-1 感染可能诱导炎症反应并影响牙周健康。
