Basic studies on adaptation the immune therapy to the patients with periodontal diseases by using synthetic peptides as an immunogen

以合成肽为免疫原对牙周病患者进行免疫治疗的基础研究

• 批准号: 09470425
• 负责人: MURAYAMA Yoji
• 金额: $ 7.68万
• 依托单位: Okayama university
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1998
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09470425/
• 关键词: Porphyromonas gingivalis; 53kDa outer membrane protein; T cell epitopes; HLA class II; Cytokines; 免疫療法; Porphyromonas gingiralis; 歯周病; ヘルパパーT細胞; HLA classII; リンフォカイン

As the first trial to adapt the immune therapy to the patients with periodontal diseases, we estimated how Ag53 is recognized by host immune cells and derive subsequent effector functions. We determined major T cell epitopes of Ag53 in EOP patients, and estimated them from the viewpoint of the restriction of HLA class II molecules. The results of this study are as follows.(1)We established Ag53 specific short-termed T cell lines from 22 subjects including 6 EOP patients and 16 healthy donors, using overlapping peptides based on Ag53 amino acid sequences. We found that all T cell lines from active EOP patients recognized common region (pl4l-p18l) on Ag53, while those of healthy donors showed heterogeneous specificity(2)Anti-DRBL monoclonal antibody inhibited Ag53-induced proliferation of most of the T cell lines.(3)A large amount of IFN-gamma was produced from all of established Th cell lines. The other cytokine production, including IL-4, 5, 6 and 10, were different among the Th cell lines.(4)The effect of the cytokine-profile produced from Th cells on the lgG production from B cells was evaluated. The Th cell lines producing high amount of Th2 type cytokines against Thl cytokines induced high IgG production. But the Th cell lines, which produced low Th2 cytokines against Thl cytokines. Did not induce the IgG production.(5)A larger amount of IL-5 from Th cells was detected in the culture with IgG production. compared to the culture without IgG production.These results suggest that AA residues from 14 1 to 181 is supposed to include major T cell epitope on Ag53 for active EOP patients but not for healthy individuals or inactive EOP patients. which might be interested in the establishment of the immune therapy for P gingivalisx infection in periodontal diseases

作为第一个将免疫疗法应用于牙周病患者的试验，我们估计了Ag53如何被宿主免疫细胞识别并衍生出后续的效应功能。我们测定了EOP患者Ag53的主要T细胞表位，并从限制HLAII类分子的角度对其进行了估计。本研究的结果如下：(1)利用基于Ag53氨基酸序列的重叠多肽，从22名受试者(包括6名EOP患者和16名健康献血者)建立了Ag53特异性短缩T细胞系。我们发现，所有活动期EOP患者的T细胞株都能识别Ag53上的共同区域(pl4l-p18l)，而健康献血者的T细胞株则表现出异质性的特异性。(2)抗DRBL单抗抑制了大多数T细胞株的增殖。IL-4、IL-5、IL-6、IL-10等细胞因子的产生在不同的Th细胞系中也不同。(4)研究了Th细胞产生的细胞因子对B细胞产生免疫球蛋白G的影响。产生高水平Th2型细胞因子对抗Th1细胞因子的Th细胞株可诱导高免疫球蛋白G产生。而产生低Th2细胞因子对抗Th1细胞因子的Th细胞系。(5)在产生免疫球蛋白的培养中，检测到大量的IL-5从Th细胞中产生。这些结果表明，活动期EOP患者的AA141~181位氨基酸残基可能包括Ag53上的主要T细胞表位，而健康人或非活动期EOP患者则不能。可能对牙周病中牙龈假单胞菌感染的免疫治疗方法的建立感兴趣

项目成果

Arai, H., et.al.: "The inhibition of DNA synthesis by prostaglandin E2 in human gingival fibroblasts is independent of the cyclic AMP-protein kinase A signal transduction pathway" Journal of Periodontal Research. 33 (1). 33-39 (1998)

Arai, H. 等人：“人牙龈成纤维细胞中前列腺素 E2 对 DNA 合成的抑制独立于环 AMP-蛋白激酶 A 信号转导途径”《牙周研究杂志》。

Oyama,H.,Murayama,Y.et al.: "T cell responses to 53-kDa outer membrane protein of Porphytomonas gingivalis in humans with early-onset neriodontitis" Human Immunology. 59(10). 635-643 (1998)

Oyama, H., Murayama, Y. 等人：“早发性神经牙炎患者中 T 细胞对牙龈卟啉单胞菌 53-kDa 外膜蛋白的反应”人类免疫学。

Sawa, T., et.al.: "In vitro induction of activation-induced cell death in lymphocytes from chronic periodontal lesion by exogeneous Fas-ligand" Infection and Immunity. 67 (3). 1450-1454 (1999)

Sawa, T., et.al.：“通过外源性 Fas 配体在体外诱导慢性牙周病灶淋巴细胞中活化诱导的细胞死亡”感染和免疫。

加藤菜保子: "Porphyromonas gingivalis 53kDa外膜蛋白を認識するT細胞のサイトカイン産生様態" 日本歯周病学会誌. 40. 144- (1997)

加藤菜穗子：“识别牙龈卟啉单胞菌 53kDa 外膜蛋白的 T 细胞的细胞因子产生模式”日本牙周病学会杂志 40. 144- (1997)。

Mizoguchi, K., et.al.: "The regulatory effect of fermentable sugar levels on the production of leukotoxin by Actinobacillus actinomycetemcomitance" FEMS Microbiology Letters. 146. 161-166 (1997)

Mizoguchi, K. 等人：“可发酵糖水平对放线杆菌放线菌产生白细胞毒素的调节作用”FEMS 微生物学快报。