Development of Rapid Rvaluation Methods for Immunopotentiators of Fish.

鱼类免疫增强剂快速评价方法的开发。

• 批准号: 08556032
• 负责人: YANO Tomoki
• 金额: $ 4.42万
• 依托单位: KYUSHU UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08556032/
• 关键词: FISH; IMMUNOPOTENTIATOR; COMPLEMENT; COMPLEMENT RECEPTOR; beta-1,3-GLUCAN; SODIUM ALGINATE; CYTOKINES; コイ; 免疫学; 補体レセプター; モノクローナル抗体

To develop rapid evaluation methods of immunopotentiators in fish, the mechanism of the immunopotentiation in carp was analyzed using a fungi-derived beta-1,3-glucan and sodium alginate prepared from a brown algae as model materials of immunopotentiators. The beta-1,3-glucan activated the complement of carp and intraperitoneal administration of the beta-1,3-glucan elevated the expression level of mammalian CR3-like complement receptors on head kidney phagocytes. Therefore, the enhancement of phagocytic activity of the phagocytes which is endowed by the increase in complement receptors expression probably plays an important role in elevation of host defense against bacterial infection. On the basis of these findings, we have isolated carp C3a fragment, which would be a good indicator for complement activation, and developed the a detection method of carp C3a as well as a rosette assay of the receptors. On the other hand, neither activation of complement nor enhancement of phagocyte activity was observed after intraperitoneally injected sodium alginate. Rather, sodium alginate induced a great migration of head kidney leukocytes to the peritoneal cavity, indicating that sodium alginate stimulate the release of chemotactic cytokines from resident leukocytes in the cavity. These results show that there are distinct mechanisms underlying the enhancing effects of those polysaccharide immunopotentiators on the fish host defense system. Functional and immunochemical assays of cytokines are not applicable to fish, because few data has been accumulated on fish cytokines. However, analysis of gene-expression, which is specifically stimulated by the administration of immunopotentiator, will help to overcome the difficulties.

为建立鱼类免疫增强剂的快速评价方法，以褐藻中提取的β-1，3-葡聚糖和海藻酸钠为免疫增强剂的模型材料，分析了鲤鱼免疫增强的机理。β-1，3-葡聚糖激活鲤鱼的补体，腹膜内给予β-1，3-葡聚糖可提高头肾吞噬细胞上哺乳动物CR 3样补体受体的表达水平。因此，补体受体表达的增加所赋予的吞噬细胞的吞噬活性的增强可能在提高宿主对细菌感染的防御中起重要作用。在此基础上，我们分离了鲤鱼补体C3 a片段，建立了鲤鱼补体C3 a的检测方法和受体花环试验。另一方面，腹腔注射海藻酸钠后，既没有观察到补体的激活，也没有观察到吞噬细胞活性的增强。相反，海藻酸钠诱导头肾白细胞大量迁移到腹腔，表明海藻酸钠刺激腔中常驻白细胞释放趋化细胞因子。这些结果表明，这些多糖免疫增强剂对鱼类宿主防御系统的增强作用存在不同的机制。细胞因子的功能和免疫化学测定不适用于鱼类，因为鱼类细胞因子的数据很少。然而，基因表达的分析，这是特别刺激免疫增强剂的管理，将有助于克服困难。

项目成果

K.Fujiki: "Protective effect of k-carrageenan against bacterial infections in carp Cyprinus carpio" J.Fac.Agr.,Kyushu Univ.42-1,2. 113-119 (1997)

K.Fujiki：“k-卡拉胶对鲤鱼细菌感染的保护作用” J.Fac.Agr.，九州大学42-1,2。

Fujiki, K., Shin, D.H., Nakao, M., and Yano, T.: "Protective effects of kappa-carrageenan against bacterial infections in carp Cyprinus carpio." J.Fac. Agr., Kyushu Univ.42(1/2). 113-119 (1997)

Fujiki, K.、Shin, D.H.、Nakao, M. 和 Yano, T.：“κ-卡拉胶对鲤鱼细菌感染的保护作用。”

K.Fujiki: "Effects of sodium alginate on the non-specific defence system of the common carp(Cyprinus carpio L.)" Fish and Shellfish Immunology. 7-6. 417-427 (1997)

K.Fujiki：“海藻酸钠对鲤鱼（Cyprinus carpio L.）非特异性防御​​系统的影响”鱼类和贝类免疫学。

Takeshi Uemura: "Purification and characterization of the eighth and ninth components of carp complement" Molecular Immunology. 33・11/12. 925-932 (1996)

Takeshi Uemura：“鲤鱼补体第八和第九成分的纯化和表征”分子免疫学33·11/12（1996）。

Miki Nakao: "Terminal components of carp complement constituting a membrane attack complex" Molecular Immunology. 33・11/12. 933-937 (1996)

Miki Nakao：“构成膜攻击复合物的鲤鱼补体的末端成分”分子免疫学33·11/12（1996）。