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Immunological relationships of NGF BDNF and NT-3: recognition and functional inhibition by antibodies to NGF

机译:NGFBDNF和NT-3的免疫学关系:NGF抗体的识别和功能抑制

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摘要

Polyclonal antibodies raised against mouse 2.5S NGF (mNGF) and against synthetic peptides made from hydrophilic portions of mNGF have been used to compare the immunological properties of mNGF, human recombinant brain-derived neurotrophic factor (hrBDNF), and human recombinant neurotrophin-3 (hrNT-3). Affinity-isolated antibodies raised against intact mNGF reacted with all three neurotrophins when tested by ELISA and totally or partially blocked the bioactivities of the proteins in survival assays of embryonic chicken sensory and sympathetic neurons. On Western blots, mNGF antibodies reacted with all three neurotrophins but less well with hrBDNF and hrNT-3 than with mNGF. Antibodies to hydrophilic peptides within NGF (amino acids 23–35, 59–67, 69–79, and 91–100) showed partial reactivity with some but not all of the neurotrophins when tested by ELISA and on Western blots. The peptide antibodies were also selectively effective in reducing the survival- promoting activity of the neurotrophins on sensory neurons. Results show that mNGF, hrBDNF, and hrNT-3 are immunologically related proteins and that mNGF antibodies react also with other members of the neurotrophin family.
机译:针对小鼠2.5S NGF(mNGF)和由mNGF亲水性部分制成的合成肽产生的多克隆抗体已用于比较mNGF,人重组脑源性神经营养因子(hrBDNF)和人重组Neurotrophin-3( hrNT-3)。当通过ELISA测试时,针对完整mNGF的亲和分离抗体与所有三种神经营养蛋白发生反应,并在胚胎鸡感觉和交感神经元的存活测定中完全或部分阻断了蛋白质的生物活性。在蛋白质印迹上,mNGF抗体与所有三种神经营养蛋白反应,但与hrBDNF和hrNT-3相比,与mNGF的反应较差。通过ELISA和Western blot检测,NGF中的亲水性肽抗体(氨基酸23–35、59–67、69–79和91–100)显示出与部分而非全部神经营养蛋白的部分反应性。肽抗体还选择性降低神经营养蛋白在感觉神经元上的存活促进活性。结果表明,mNGF,hrBDNF和hrNT-3是免疫相关蛋白,mNGF抗体也与神经营养蛋白家族的其他成员发生反应。

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