Akhter Firoz, Khan M Salman, Ahmad Saheem
Department of Bio-Sciences, Integral University, Lucknow, 226026, India; Department of Bio-Engineering, Integral University, Lucknow, 226026, India.
Department of Bio-Sciences, Integral University, Lucknow, 226026, India.
Int J Biol Macromol. 2015 Jan;72:1222-7. doi: 10.1016/j.ijbiomac.2014.10.034. Epub 2014 Oct 26.
Glycation of biologically important macromolecules leads to the establishment of advanced glycation end products (AGEs) having significant role in the pathophysiology of various diseases. d-Ribose, is a highly reactive pentose sugar resulting in the rapid formation of AGEs. Formation of d-ribose derived glycated DNA and LDL has been previously demonstrated; however no comparative, extensive studies have been performed to assess the immunogenicity of d-ribose glycated calf thymus DNA (CT-DNA) and LDL. In the present study, the results showed that animals immunized with d-ribose modified CT-DNA and LDL induced antibodies as detected by direct binding and competition ELISA. The modified CT-DNA and LDL were found to be highly immunogenic, eliciting high titer immunogen-specific antibodies, while the native forms of DNA was almost non-immunogenic. The induced antibodies from modified CT-DNA and LDL exhibited wide range of heterogeneity in recognizing various nucleic acid conformers, DNA bases and amino acids. Furthermore, Serum antibodies from diabetes and diabetes atherosclerosis patients were screened for their binding to native CT-DNA, LDL and glycated CT-DNA, LDL. Glycated CT-DNA showed almost equivalent binding to both diabetes and diabetic atherosclerosis group while high recognition was observed when glycated LDL was used as an antigen.
生物重要大分子的糖基化会导致晚期糖基化终产物(AGEs)的形成,这些产物在多种疾病的病理生理学中具有重要作用。d-核糖是一种高反应性的戊糖,会迅速形成AGEs。此前已证明d-核糖可导致糖化DNA和低密度脂蛋白(LDL)的形成;然而,尚未进行比较广泛的研究来评估d-核糖糖化小牛胸腺DNA(CT-DNA)和LDL的免疫原性。在本研究中,结果表明,用d-核糖修饰的CT-DNA和LDL免疫的动物,通过直接结合和竞争酶联免疫吸附测定(ELISA)检测到诱导产生了抗体。发现修饰后的CT-DNA和LDL具有高度免疫原性,可引发高滴度的免疫原特异性抗体,而天然形式的DNA几乎没有免疫原性。修饰后的CT-DNA和LDL诱导产生的抗体在识别各种核酸构象、DNA碱基和氨基酸方面表现出广泛的异质性。此外,还筛选了糖尿病和糖尿病动脉粥样硬化患者血清抗体与天然CT-DNA、LDL以及糖化CT-DNA、LDL的结合情况。糖化CT-DNA与糖尿病组和糖尿病动脉粥样硬化组的结合几乎相当,而以糖化LDL作为抗原时则观察到更高的识别率。