Idiotypic analysis of serum anti-A. actinomycetemcomitans antibody from penodontitis patients

血清抗 A 的独特型分析。

• 批准号: 09671946
• 负责人: NAKASHIMA Keisuke
• 金额: $ 1.98万
• 依托单位: Health Sciences University of Hokkaido
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1997
• 资助国家: 日本
• 起止时间: 1997 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09671946/
• 关键词: 歯周病原性細菌; 患者血清; イディオタイプ

In 1997 and 1998, we tried to establish human-mouse heterohybridomas producing antibodies to the serotype-b specific antigen from Actinobacillus actinomycetemcomitans in order to analyze idiotype of the antibodies. However, none of the twenty established hybridomas showed positive reactions with the serotype-b specific antigen. In 1999, we tried idiotypic analysis of the mouse monoclonal antibody to the serotype-b specific antigen by preparing single-chain F-. Variable (ScFv) recombinant antigen-binding molecules. mRNA was isolated from two hybridomas (S5; specific for the serotype-b specific antigen, L2; specific for LPS from A. actinomycetemcomitans), and cDNAs of variable regions of H- and L-chains were amplified by RT-PCR. cDNA could not be amplified as for the L-chain of Mab S5, although we confirmed that the L-chain is kappa chain. Amplified cDNAs of H- and L-chains of Mab L2 were joined into a single chain (ScFv) with linker DNA. The ScFv was ligated to pGEM T-easy vector, and transformed to JM 109 competent cell. Two clones selected by insert check were analyzed for DNA sequence. In conclusion, we could not have the expected results in this study, however, the DNA sequence obtained in this study would be an important tool to proceed idiotypic analysis of serum anti-A. actinomycetemcomitans antibody.

在1997年和1998年，我们试图建立人-鼠异源杂交瘤，产生抗放线杆菌B型特异性抗原的抗体，以分析抗体的独特型。然而，20个建立的杂交瘤细胞中没有一个显示出与B型特异性抗原的阳性反应。1999年，我们尝试了用单链F-抗体制备抗B型特异性抗原的鼠单克隆抗体进行独特型分析。可变（ScFv）重组抗原结合分子。从两个杂交瘤中分离mRNA（S5;对B型特异性抗原特异，L2;对来自A.用RT-PCR方法扩增了放线菌伴放线菌（actinomycetemcomitans）的H链和L链可变区的cDNA。虽然我们证实了单克隆抗体S5的L-链是κ链，但不能扩增出与L-链相同的cDNA。用接头DNA将Mab L2的H链和L链的扩增cDNA连接成单链（ScFv）。将单链抗体连接到pGEM T-easy载体上，转化JM 109感受态细胞。分析通过插入检查选择的两个克隆的DNA序列。总之，本研究未能获得预期的结果，但本研究获得的DNA序列将为血清抗A独特型分析提供重要工具。伴放线菌抗体

项目成果

K.NAKASHIMA: "Two different types of humoral immune response to Actinobacilllus actinomycetemcomitans in high-responder periodontitis patients" J.Med.Microbiol. 47. 569-575 (1998)

K.NAKASHIMA：“高反应性牙周炎患者对伴放线杆菌的两种不同类型的体液免疫反应”J.Med.Microbiol。

Nakashima K.: "Two different types of humoral immune response to A. actinomycetemcomitans in high-responder periodontitis patients"Journal of Medical Microbiology. 47. 569-575 (1998)

Nakashima K.：“高反应性牙周炎患者对伴放线菌的两种不同类型的体液免疫反应”医学微生物学杂志。