The identification of the disulfide bonds in HIV gp120 whose reduction is required for cell entry and their manipulation for immunogen design

HIV gp120 中二硫键的鉴定（其还原是细胞进入所需的）及其用于免疫原设计的操作

• 批准号: MR/J008796/1
• 负责人: Ian Jones
• 金额: $ 63万
• 依托单位: University of Reading
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2012
• 资助国家: 英国
• 起止时间: 2012 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FJ008796%2F1
• 关键词: identification; disulfide; bonds; HIV; gp120

Infection by the Human immunodeficiency virus (HIV) is the cause of acquired immunodeficiency syndrome (AIDS) and is a major cause of death worldwide. Despite extensive research efforts the search for an effective vaccine has not yet been successful and there is a continued need for new candidates to be assessed. The virus enters the cell via interaction between the virus envelope glycoprotein and the target cell surface, in particular two cell surface molecules called CD4 and CCR. Following contact between these molecules a number of biochemical steps occur which result in the virus penetrating the cell to start the infection. The precise nature of the biochemical steps required is still a matter of research but if they are identified clearly they may be useful in the design of the new potential vaccines. We have discovered one particular biochemical change, disulfide bond reduction, which appears to be important for virus entry. If this step is inhibited virus infection cannot occur suggesting it is a key intermediate stage in the entry process. We want to use a new technology involving very sensitive mass spectrometry to identify the particular disulfide bonds of the virus envelope glycoprotein that are reduced during the infection process in order to provide a greater level of clarity about the entry process than is currently the case. When the particular disulfide bonds are known we will make some of the reduced protein and compare its properties to the normal, unreduced form. We will also test if this intermediate protein is a key part of the HIV infection process by using it to generate antibodies which will be tested to see if they prevent infection. Antibodies, the host's response to infection, which bind to the reduced protein and stop its function will prove that the reduced protein can be considered a valid vaccine candidate.

人类免疫缺陷病毒（HIV）感染是获得免疫缺陷综合征（AIDS）的原因，是全球死亡的主要原因。尽管进行了广泛的研究工作，但寻找有效的疫苗尚未成功，并且需要对新候选人进行评估。该病毒通过病毒包膜糖蛋白和靶细胞表面，尤其是两个称为CD4和CCR的细胞表面分子之间的相互作用进入细胞。在这些分子之间接触后，发生了许多生化步骤，导致病毒穿透细胞开始感染。所需的生化步骤的确切性质仍然是研究问题，但是如果清楚地识别出它们，它们可能在新的潜在疫苗的设计中有用。我们发现了一种特殊的生化变化，二硫键降低，这对于进入病毒似乎很重要。如果此步骤被抑制，则不会发生病毒感染，这表明它是进入过程中的关键中间阶段。我们希望使用涉及非常敏感的质谱法的新技术来识别病毒信封糖蛋白的特定二硫键键，该糖蛋白在感染过程中降低，以便比目前的情况更明确。当知道特定的二硫键键时，我们将制造一些还原的蛋白质，并将其特性与正常的未还原形式进行比较。我们还将通过使用该抗体生成抗体来测试该中间蛋白是否是艾滋病毒感染过程的关键部分，以查看它们是否防止感染。抗体，宿主对感染的反应，结合了降低的蛋白质并停止其功能，将证明降低的蛋白质可以视为有效的疫苗候选者。

项目成果

Intracellular Trafficking, Localization, and Mobilization of Platelet-Borne Thiol Isomerases.

• DOI: 10.1161/atvbaha.116.307461
• 发表时间: 2016-06
• 期刊: Arteriosclerosis, thrombosis, and vascular biology
• 作者: Crescente M;Pluthero FG;Li L;Lo RW;Walsh TG;Schenk MP;Holbrook LM;Louriero S;Ali MS;Vaiyapuri S;Falet H;Jones IM;Poole AW;Kahr WH;Gibbins JM
• 通讯作者: Gibbins JM

OX133, a monoclonal antibody recognizing protein-bound N-ethylmaleimide for the identification of reduced disulfide bonds in proteins.

• DOI: 10.1080/19420862.2016.1152443
• 发表时间: 2016-05
• 期刊: mAbs
• 影响因子: 5.3
• 作者: Holbrook LM;Kwong LS;Metcalfe CL;Fenouillet E;Jones IM;Barclay AN
• 通讯作者: Barclay AN

The production, characterisation and application of monoclonal antibodies generated by immunisation with HIV-1C clade RGP140 envelope protein.

HIV-1C 进化枝 RGP140 包膜蛋白免疫产生的单克隆抗体的生产、表征和应用。

• DOI: 10.1016/j.jviromet.2013.08.011
• 发表时间: 2013
• 期刊: Journal of virological methods
• 影响因子: 3.1
• 作者: Hassall M