L-天冬酰胺酶表面呈現對P277肽免疫原性及抗NOD小鼠糖尿病作用的影響

Effect of Displaying P277 Peptide on Surface of L-asparaginase on Its Antigenicity and Efficacy of Autoimmune Diabetes Prevention in NOD Mice

doi：

摘要：

將P277多肽融合在L-天冬酰胺酶的C端,中間引入破傷毒素肽(TTP),重組嵌合酶AnsB-TTP-P277能以可溶性蛋白的形式表達于大腸桿菌周質內,且能保持大約80%的天然酶催化活力.用純化的嵌合酶腹腔注射非肥胖性(NOD)小鼠,可成功誘導小鼠體內產生高水平抗P277抗體.研究表明,P277肽可呈現于L-天冬酰胺酶表面,有效地克服單獨P277的弱免疫原性,激發機體產生較強烈的免疫反應.小鼠胰腺病理學切片也顯示:重組嵌合酶AnsB-TTP-P277和單純P277肽相比,能更有效地抑制NOD小鼠糖尿病的發生.

Abstract：

The recombinant chimeric enzyme of AnsB-TTP-P277 comprising L-asparaginase, a tetanus toxin peptide spacer and P277 was expressed as a soluble protein in Escherichia coli. The purified chimeric enzyme exhibited primary activity of the native asparaginase. Prediabetic NOD mice immunized with the chimeric enzyme could induce specific antibodies against P277 and the specificity of anti-P277 antibodies was verified by Western blot assay. The study showed that displaying the P277 epitope on the surface of asparaginase could effectively overcome the weak antigenicity of the P277 epitope and evoke a strong P277-specific immune response in mice. Moreover, the concentration of blood glucose was measured by an automated analyzer.Histochemical analysis of mice pancreas tissues showed that the administration of the chimeric enzyme to NOD mice could prevent the development of diabetes more efficiently than the peptide P277 itself.

作者：	朱愛華[1]劉文濤[2]龍軍[2]吳潔[2]劉景晶[2]李泰明[2]
Author：	ZHU Ai-Hua[1]  LIU Wen-Tao[2]  Long Jun[2]  Wu Jie[2]  LIU Jing-Jing[2]  LI Tai-Ming[2]
作者單位：	中國藥科大學生命科學與技術學院微基因藥物實驗室,南京,210009;徐州師范大學生命科學學院,江蘇,徐州,221116 中國藥科大學生命科學與技術學院微基因藥物實驗室,南京,210009
期 刊：	中國生物化學與分子生物學報   ISTICPKU
Journal：	CHINESE JOURNAL OF BIOCHEMISTRY AND MOLECULAR BIOLOGY
年，卷(期)：	2007, 23(9)
分類號：	Q789 R392
關鍵詞：	L-天冬酰胺酶    P277肽    胰島素依賴型糖尿病    抗體    血糖
Keywords：	L-asparaginase    P277 peptide    insulin-dependent diabetes mellitus    antibody    blood glucose
機標分類號：	R73 R57
機標關鍵詞：	天冬酰胺酶    表面呈現    免疫原性    小鼠胰腺    糖尿病    Escherichia coli    soluble protein    immune response    blood glucose    Western blot    酶催化活力    可溶性蛋白    重組    形式表達    小鼠體內    免疫反應    腹腔注射    大腸桿菌    非肥胖    病理學
基金項目：	國家高技術研究發展計劃(863計劃)，國家自然科學基金