Design and characterization of epitope-scaffold immunogens that present the motavizumab epitope from respiratory syncytial virus.
Design and characterization of epitope-scaffold immunogens that present the motavizumab epitope from respiratory syncytial virus.
复制标题
呼吸道合胞病毒的表位体外免疫原子的设计和表征。
DOI：
10.1016/j.jmb.2011.04.044
复制
发表时间：
2011-06-24
影响因子：
5.6
通讯作者：
Kwong PD
中科院分区：
文献类型：
--
作者：
McLellan JS;Correia BE;Chen M;Yang Y;Graham BS;Schief WR;Kwong PD
呼吸道合胞病毒（RSV）是婴儿呼吸道感染的主要病因，但目前尚未开发出有效的疫苗。一种理想的疫苗应能激发保护性抗体，同时避免病毒特异性T细胞反应，此前的RSV疫苗中已发现这种反应与疫苗增强性疾病有关。我们提出，设计用于呈现RSV中和抗体表位并激发同源抗体的异源蛋白有可能满足这些疫苗要求，因为它们可以被设计成不含病毒T细胞表位。在此，我们介绍了三种表位支架的设计和特性，它们呈现莫他珠单抗的表位，莫他珠单抗是一种能与RSV融合（F）糖蛋白中的螺旋 - 环 - 螺旋基序结合的强效中和抗体。其中两种表位支架能够被纯化，一种基于金黄色葡萄球菌A蛋白结构域的表位支架与莫他珠单抗结合，其动力学和热力学性质表明，游离的表位支架在一种与莫他珠单抗结合状态非常相似的构象中稳定下来。通过圆二色性和等温滴定量热法评估，该表位支架折叠良好，其与莫他珠单抗形成的复合物的晶体结构分辨率达到1.9 Å，与计算机设计的模型相似，所有对与莫他珠单抗结合至关重要的氢键相互作用都得以保留。用这种表位支架免疫小鼠未能激发中和抗体，但确实激发了具有F结合活性的血清。F结合抗体的激发表明，在激发保护性抗体且无病毒特异性T细胞反应的一些设计标准正在得到满足，但这些新型免疫原还需要进一步优化。
Respiratory syncytial virus (RSV) is a major cause of respiratory tract infections in infants, but an effective vaccine has not yet been developed. An ideal vaccine would elicit protective antibodies while avoiding virus-specific T-cell responses, which have been implicated in vaccine-enhanced disease with previous RSV vaccines. We propose that heterologous proteins designed to present RSV neutralizing-antibody epitopes and to elicit cognate antibodies have the potential to fulfill these vaccine requirements, as they can be fashioned to be free of viral T-cell epitopes. Here we present the design and characterization of three epitope-scaffolds that present the epitope of motavizumab, a potent neutralizing antibody that binds to a helix-loop-helix motif in the RSV fusion (F) glycoprotein. Two of the epitope-scaffolds could be purified, and one based on a S. aureus Protein A domain bound motavizumab with kinetic and thermodynamic properties consistent with the free epitope-scaffold being stabilized in a conformation that closely resembled the motavizumab-bound state. This epitope-scaffold was well-folded as assessed by circular dichroism and isothermal titration calorimetry, and its crystal structure – determined in complex with motavizumab to 1.9 Å resolution – was similar to the computationally-designed model, with all hydrogen-bond interactions critical for binding to motavizumab preserved. Immunization of mice with this epitope-scaffold failed to elicit neutralizing antibodies, but did elicit sera with F-binding activity. The elicitation of F-binding antibodies suggests that some of the design criteria to elicit protective antibodies without virus-specific T-cell responses are being met, but additional optimization of these novel immunogens is required.
登录
查看更多内容
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
Emsley, P;Cowtan, K
通讯作者：
Cowtan, K
DOI：
10.1056/nejmoa0804877
发表时间：
2009-02-05
期刊：
The New England journal of medicine
影响因子：
--
作者：
Hall CB;Weinberg GA;Iwane MK;Blumkin AK;Edwards KM;Staat MA;Auinger P;Griffin MR;Poehling KA;Erdman D;Grijalva CG;Zhu Y;Szilagyi P
通讯作者：
Szilagyi P
DOI：
10.1073/pnas.97.19.10383
发表时间：
2000-09-12
影响因子：
11.1
作者：
Kuhlman, B;Baker, D
通讯作者：
Baker, D
DOI：
10.1073/pnas.1004728107
发表时间：
2010-10-19
影响因子：
11.1
作者：
Ofek, Gilad;Guenaga, F. Javier;Kwong, Peter D.
通讯作者：
Kwong, Peter D.
影响因子：
2.2
作者：
Chen, M.;Chang, J. S.;Nason, M.;Rangel, D.;Gall, J. G.;Graham, B. S.;Ledgerwood, J. E.
通讯作者：
Ledgerwood, J. E.