Induction of heat shock protein and gamma/delta T cells in peritoneal exudate cells of mice infected with periodontopathic bacteria

本研究旨在探讨宿主对牙周病细菌的局部反应在诱导热休克蛋白（HSP）、γ / δ T细胞、细胞因子和巨噬细胞活化中的作用。我们选用放线菌（放线菌）、牙龈卟啉单胞菌（Pg）和核梭杆菌（Fn）作为牙周病原菌。分别以1/10 MLD的剂量腹腔注射，按0、1、3、5、7、14 d的时间过程观察宿主反应。在免疫方面，小鼠每周注射3次1/10 MLD活菌，分别腹腔内和皮下注射。末次注射后10 d作为免疫小鼠。实验结果如下：1.实验结果表明：注射Fn后，小鼠腹膜渗出细胞（PEC）提取物中hsp70和60表达升高。Fn注射组60 HSP增加1.6倍，70HSP增加2.6倍。注射Fn后γ /deltaT细胞百分比增加，在第7天达到27%的峰值。注射Aa或Pg均未诱导出γ /deltaT细胞，但有少量γ /deltaT细胞。采用小鼠细胞因子ELISA试剂盒（ENDOGEN）检测细菌注射后细胞因子水平。腹腔炎症细胞因子在注射Aa、Pg和Fn后12 h达到峰值，随即下降。腹腔内ifn - γ和IL-12水平较高。注射Aa后血清IL-4水平升高，在第7天达到较高水平。3 .所有细菌的局部IK-4水平均未升高。Fn免疫诱导巨噬细胞活化。综上所述，Aa和Pg诱导免疫后的炎症动因子、α / β细胞、IL-4和IgG抗体的产生，而Fn可能导致免疫后HSP、γ / δ t细胞和巨噬细胞的顺序活化。

The purpose of this study was to examine the local reaction of host against periodontopathic bacteria in induction of heat shock protein (HSP), gamma/delta T cells, cytokine and macrophage activation. We used as periodontal pathogens, Actinobacillus actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg) and Fusobacterium nucleatum (Fn). They were injected intraperitoneally at the dose of 1/10 MLD ant the host reaction was observed according to the the time course of 0,1,3,5,7 and 14 days. For immunization, mice were injected with 1/10 MLD of live bacteria 3 times in every week, intraperitoneally intraperitoneally and subcutaneously. They were used as immunized mouse, 10 days after last injection. The results were as follows :1. Increase of HSP 70 and 60 were found in the extract of peritoneal exudate cells (PEC) from mice which received the Fn injection. An Increase of 1.6 times in 60 HSP,2.6 times in 70HSP was observed in the case of Fn injection.2. Increase of gamma/deltaT cells in percentage after injection of Fn was observed showing the peak of 27% on 7 day. Induction of gamma/deltaT cells was not occurred by the injection of Aa or Pg showing several parcent of gamma/deltaT cells.3. The cytokine level was measured after injection of the bacteria using the mouse cytokine ELISA kit (ENDOGEN). Inflamatory cytokines in the abdomen reached the peak at 12 h after injection of Aa, Pg and Fn, and decreased immediately. IFN-gamma and IL-12 levels in the abdomen were high in the abdominal cavity. IL-4 in serum was increased and showed a high level on Day 7 by injection of Aa. The local level of IK-4 was not elevated in any bacteria.4. Activation of macrophages was induced by Fn immunization. In conclusion, Aa and Pg induced inflamatory dytokine, alpha/betaT cells, IL-4 and IgG antibody production, whereas Fn may result in the sequential appearance of HSP,gamma/delta Tcell and macrophage activation after immunization.