| Summary: | 碩士 === 國立陽明大學 === 口腔生物研究所 === 99 === Type 1 herpes simplex virus (HSV-1) is a common infectious agent. It could cause inflammation at the site of infection. Neutrophils are important immune cells at the site of inflammation. Previous studies have shown that HSV-1-immune complexes are formed by HSV-1 opsonized by complement or anti-HSV-1 antibody. Neutrophils are then able to bind and internalize HSV-1-immune complexes and ultimately lead to degradation of viruses. Other studies have shown that neutrophils could bind to HSV-1 directly. On the other hand, HSV-1 could neither replicate in neutrophils, nor induce adult neutrophils apoptosis. The effects of HSV-1 infection on the functions of neutrophils have not been well studied. The purpose of the study was to examine if HSV-1 could bind to neutrophils and to determine the effects of HSV-1 infection on the functions of neutrophils. HSV-1 was pretreated with the HSV-1-antiserum to make HSV-1-immune complexes. The binding and internalization of HSV-1 by neutrophils and the formation of endosome were observed by immunostanning. The viability of neutrophils was determined using the trypan blue dye exclusion test. The cellular reactive oxygen species (ROS) levels in neutrophils were determined by the ROS test. Production of interleukin-8 (IL-8) and leukotriene B4 (LTB4) was measured by ELISA (enzyme-linked immunosorbent assay) and EIA (enzyme immunoassay), respectively. Matrix metalloproteinase-9 (MMP-9) was measured using zymography. Production of viral proteins and the activation of phosphatidylinositol-3 kinase (PI3K) of neutrophils were measured using Western blot analysis. The results showed that HSV-1 could bind and enter neutrophils in the absence of HSV-1-antiserum. The binding effect of HSV-1-immune complexes was more evident. The viability of neutrophils was not affected by HSV-1 infection in three hours. HSV-1-immune complexes elevated the internal ROS levels, activated the PI3K pathway, and also stimulated neutrophils to release mediators, IL-8 and LTB4, significantly. Secretion of MMP-9 was elevated when neutrophils were infected by HSV-1. The results in this study showed that HSV-1 and HSV-1-immune complexes activated neutrophils differently, which indicated that neutrophils played an important role in the defense of HSV-1 infection in the presence or absence of HSV-1-antiserum.
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