[Mechanism of stimulation of spleen mononuclear cells by gram-positive bacterial peptidoglycan].

M Sashida
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Abstract

Periodontal disease is a chlonic inflammatory disorder, and for which oral microbes are supposed to be responsible. Among oral microbials, gram-negative bacterias have been studied extensively in relation to periodontal disease for their pathogenicity due to their lipopolysaccharide (LPS), exocellular enzymes or bacterial toxin. As for gram-positive bacterials, it has been reported recently that gram-positive bacteria can elicit immunological responses, and this may be responsible for the initiation and/or development of periodontal disease. However, precise mechanisms of bacterial action, especially of gram-positive bacteria, on periodontal disease have not been elucidated yet. In this experiment, therefore, gram-positive bacteria (S. epidermidis), peptidoglycan subunits of S. epidermidis (SEPS) and muramyl dipeptide (MDP) were used to investigate for their activities to stimulate spleen mononuclear cells to replicate and produce various kinds of cytokines. Immunological responsibilities of various strains of mice were explored to investigate the difference of defence of mechanisms. Following results were obtained. (1) S. epidermidis itself showed a extremely low cell-mediated activity to stimulate the replication of spleen mononuclear cells in contrast to E. coli. Staphylococcal phage lysate and SEPS stimulated remarkally the replication of spleen mononuclear cells. (2) The stimulation of spleen mononuclear cells was accompanied by the production of interleukin 3 (IL-3) and colony stimulating factor (CSF), but interleukin 2(IL-2) was not produced as in the case of E. coli. (3) Analysis of cell surface antigens revealed the increase of the numbers of Ia+ and Mac-2+ bone marrow cells following stimulation of spleen mononuclear cells with SEPS. However, T or B cells were not increased. (4) Macrophage-depleted and antisera Ia-treated spleen mononuclear cells showed a marked decrease of replicating activity of spleen mononuclear cells. (5) Among the various strains of mice tested C3H/HeN, Balb/c, AKR, DBA/2, C57BL/6, ddY, C3H/HeJ, MRL/lpr and showed a high immunological responses, but Balb/c did not. C3C/HeJ and MRL/lpr also lacked immunological reactivity. These results suggest that proliferative response of lymphocyte with peptidoglycan in gram-positive bacterium is very important for infection and its defensive reaction against gram-positive bacteria.

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革兰氏阳性细菌肽聚糖刺激脾单核细胞的机制。
牙周病是一种慢性炎症性疾病,口腔微生物被认为是罪魁祸首。在口腔微生物中,革兰氏阴性菌由于其脂多糖(LPS)、胞外酶或细菌毒素的致病性而与牙周病的关系被广泛研究。至于革兰氏阳性细菌,最近有报道称革兰氏阳性细菌可以引起免疫反应,这可能是牙周病发生和/或发展的原因。然而,细菌作用的确切机制,特别是革兰氏阳性细菌,在牙周病尚未阐明。因此,本实验以革兰氏阳性菌表皮葡萄球菌(S. epidermidis)、表皮葡萄球菌肽聚糖亚基(SEPS)和muramyl二肽(MDP)为研究对象,研究其刺激脾单核细胞复制并产生多种细胞因子的活性。探讨不同品系小鼠的免疫功能,探讨其防御机制的差异。得到以下结果:(1)与大肠杆菌相比,表皮葡萄球菌本身表现出极低的细胞介导活性来刺激脾脏单核细胞的复制。葡萄球菌噬菌体裂解液和SEPS对脾单核细胞的复制有显著的刺激作用。(2)脾脏单核细胞受到刺激时,会产生白细胞介素3 (IL-3)和集落刺激因子(CSF),但不像大肠杆菌那样产生白细胞介素2(IL-2)。(3)细胞表面抗原分析显示,SEPS刺激脾脏单核细胞后,骨髓Ia+和Mac-2+细胞数量增加。但T细胞和B细胞均未增加。(4)巨噬细胞缺失和抗血清ia处理的脾脏单核细胞复制活性明显降低。(5)各品系小鼠C3H/HeN、Balb/c、AKR、DBA/2、C57BL/6、ddY、C3H/HeJ、MRL/lpr均表现出较高的免疫应答,而Balb/c则表现出较低的免疫应答。C3C/HeJ和MRL/lpr也缺乏免疫反应性。这些结果提示,革兰氏阳性菌中淋巴细胞对肽聚糖的增殖反应对感染及其对革兰氏阳性菌的防御反应具有重要意义。
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