Scragg M A, Turton L R, Williams D M
Department of Oral Pathology, London Hospital Medical College, England.
Arch Oral Biol. 1990;35(3):167-73. doi: 10.1016/0003-9969(90)90051-b.
This study aimed to determine whether leucocytes exposed to Bacteroides gingivalis culture supernatant exhibited consistent morphological changes and whether measurement of such changes could provide a simple screening assay for bacterial products with potential biological activity. Glass-adherent polymorphonuclear leucocytes were obtained from clotted blood preparations and from blood after purification by centrifugation through Ficoll-Hypaque. These were then incubated in Hanks' balanced salt solution, a sterile liquid medium (BM) and B. gingivalis (W83) culture supernatant. Polymorphonuclear leucocytes were classified by their shape into small non-polar (less than 18 microns diameter), large non-polar (greater than or equal to 18 microns diameter), bipolar and hyperpolar types. Treatment with B. gingivalis culture supernatant consistently increased large non-polar cells by 150% to over 300% (p less than 0.01), when compared with polymorphonuclear leucocytes incubated with the sterile liquid medium (control). This change was accompanied by smaller decreases in small non-polar, bipolar and hyperpolar cells, these being significant for bipolar cells in clot preparations (p less than 0.01) and small non-polar cells after Ficoll-Hypaque isolation (p less than 0.01). Neither the liquid medium nor the B. gingivalis culture supernatant was toxic to the cells as indicated by trypan blue exclusion tests. Lipopolysaccharide and short-chain fatty acids were not responsible for the changes in polymorphonuclear leucocyte shape. However, the activity of the culture supernatant was destroyed by heating at 80 degrees C for 30 min, indicating that proteolytic enzymes may have been involved.
本研究旨在确定暴露于牙龈卟啉单胞菌培养上清液的白细胞是否呈现出一致的形态变化，以及测量这些变化是否能够为具有潜在生物活性的细菌产物提供一种简单的筛选检测方法。玻璃黏附多形核白细胞取自凝血制剂以及经Ficoll-Hypaque离心纯化后的血液。然后将这些细胞在汉克斯平衡盐溶液、无菌液体培养基（BM）以及牙龈卟啉单胞菌（W83）培养上清液中进行孵育。多形核白细胞根据其形状分为小的非极性细胞（直径小于18微米）、大的非极性细胞（直径大于或等于18微米）、双极细胞和超极细胞。与在无菌液体培养基（对照）中孵育的多形核白细胞相比，用牙龈卟啉单胞菌培养上清液处理后，大的非极性细胞持续增加了150%至超过300%（p<0.01）。这种变化伴随着小的非极性细胞、双极细胞和超极细胞数量的较小减少，对于凝血制剂中的双极细胞（p<0.01）以及Ficoll-Hypaque分离后的小的非极性细胞（p<0.01）而言，这些减少具有统计学意义。台盼蓝排斥试验表明，液体培养基和牙龈卟啉单胞菌培养上清液对细胞均无毒性。脂多糖和短链脂肪酸并非多形核白细胞形状变化的原因。然而，培养上清液的活性在80℃加热30分钟后被破坏，这表明可能涉及蛋白水解酶。
