伴放线放线杆菌诱导人外周血淋巴细胞活化及凋亡的体外研究

目的研究伴放线放线杆菌诱导人外周血淋巴细胞活化及凋亡的作用。方法选取10名全身及牙周组织健康受试者，分离外周血淋巴细胞，在有／无伴放线放线杆菌情况下培养0—96h，用荧光探针（AnnexinV—FITC、PI、CD69-TC7）进行标记，并进行流式细胞仪检测。结果全淋巴细胞加伴放线放线杆菌组AnnexinV＋／PI-细胞百分数在48h、72h、96h分别为13．42±2．88、22．74±2．18、46．92±4．28，全淋巴细胞组AnnexinV＋／PI-细胞百分数在48h、72h、96h分别为8．46±2．53、6．36±2．36、9．36±2．67，2组间存在明显差异（P〈0．01）。CD69加淋巴细胞加伴放线放线杆菌组和CD69＋淋巴细胞组AnnexinV＋／PI-细胞百分数除48h外的4个时间点上都无明显差异（P〉0．05）。CD69＋淋巴细胞加伴放线放线杆菌组AnnexinV＋／PI-细胞百分数在各个时间点上都明显高于全淋巴细胞加伴放线放线杆菌组（P〈0．01）。结论伴放线放线杆菌能够诱导人外周血淋巴细胞活化，并且能够通过活化促进其凋亡。

In vitro study of peripheral blood lymphocytes activation and apoptosis induced by Actinobacillus actinomycetemcomitans

Objective To investigate the induction effects of Actinobacillus actinomycetemcomitans on activation and apoptosis of the human peripheral blood lymphocytes. Methods Peripheral blood lymphocytes of healthy volunteers were isolated by Ficoll-Hypaque density gradient centifugation. Then the cells were incubated with or without over 0 - 96 h and the incubation was stopped at 0,24,48,72,96 hours, respectively. The cells were labeled with fluorochrome-conjugated monoclonal antibodies and probes （Annexin V-FITC, PI, CD69-TC7 ） and detected with flow cytometry. Results The percentages of Annexin V ＋/PI- cells in the group of whole lymphocytes with A. actinomycetemcomitans were 13.42 ± 2. 88, 22. 74 ± 2. 18, 46. 92 ± 4. 28 at 48h,72h,96h respectively; and those in the group of whole lymphocytes without A. actinomycetemcomitans were 81 46± 2. 53,6. 36 ± 2. 36,9. 36 ± 2. 67 respectively. The differences in percentages of Annexin V ＋/PI- cells between the two groups were significant （ P 〈 0. 01 ） . The percentages of Annexin V ＋/PI- cells in the group of CD69 ＋ lymphocytes with A. actinomycetemcomitans and the group without A. actinomycetemcomitans were not significantly different （ P 〉 0. 05 ） at four time points except at 48 h. The percentages of Annexin V ＋/PI- cells in the group of whole lymphocytes with A. actinomycetemcomitans were greater thanthose in the group of CD69 ＋ lymphocytes with A. actinomycetemcomitans （P〈0. 01 ） . Conclusion A. actinomycetemcomitans germ cell could induce the activation of lymphocytes and promote the apoptosis throuth activation.