Dense Bodies of a gH/gL/UL128/UL130/UL131 Pentamer-Repaired Towne Strain of Human Cytomegalovirus Induce an Enhanced Neutralizing Antibody Response
Dense Bodies of a gH/gL/UL128/UL130/UL131 Pentamer-Repaired Towne Strain of Human Cytomegalovirus Induce an Enhanced Neutralizing Antibody Response
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DOI：
10.1128/jvi.00931-19
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发表时间：
2019-09-01
影响因子：
5.4
通讯作者：
Plachter, Bodo
中科院分区：
文献类型：
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作者：
Lehmann, Caroline;Falk, Jessica Julia;Plachter, Bodo
针对人巨细胞病毒感染（HCMV）的疫苗的开发是高度优先的医学目标。由糖蛋白H（gH）/gL/UL 128 - 131 A（PC）组成的病毒五聚体蛋白复合物被认为是重要的疫苗组分。然而，它与诱导保护性抗体反应的相关性仍然是一个有争议的问题。我们通过使用HCMV的亚病毒致密体（DBs）解决了这个问题。DB具有异常的免疫原性。实验室HCMV毒株DB具有重要的中和抗体靶标，如糖蛋白B、H、L、M和N，但它们缺乏PC。为了能够直接比较PC对中和抗体（NT-abs）应答水平的影响，通过细菌人工染色体（BAC）诱变（Towne-UL 130 rep）建立了HCMV实验室菌株Towne的PC阳性变体。该菌株在感染成纤维细胞后合成PC阳性DB。这些DB与PC阴性Towne DB一起用于并行免疫。检测小鼠和兔血清，以确定PC对DB免疫原性的影响。对成纤维细胞、上皮细胞和内皮细胞以及使用的两种动物种属进行的检测显示，对PC阳性DB的中和抗体应答上级优于PC阴性DB。实验揭示了PC增强针对HCMV的抗体应答的潜力。特别令人感兴趣的是，发现PC阳性DB诱导的抗体反应，阻止PC阳性病毒株感染成纤维细胞比血清更有效地与PC阴性particle.IMPORTANCE免疫接种后，感染人巨细胞病毒（HCMV）可能会导致严重的，甚至危及生命的疾病表现在新生儿和免疫抑制的个人。目前正在采取几种策略来开发针对这种病毒的疫苗。这方面的一个关键问题是成功疫苗的抗原组成。使用亚病毒颗粒疫苗候选者，我们在这里表明，HCMV的一种蛋白质复合物，称为五聚体复合物（PC），增强了对不同细胞类型病毒感染的中和抗体应答。我们还首次表明，这不仅与上皮细胞或内皮细胞的感染有关;颗粒中PC的存在还增强了针对HCMV感染成纤维细胞的中和抗体应答。总之，这些发现支持将PC纳入HCMV疫苗开发策略。
The development of a vaccine against human cytomegalovirus infection (HCMV) is a high-priority medical goal. The viral pentameric protein complex consisting of glycoprotein H (gH)/gL/UL128-131A (PC) is considered to be an important vaccine component. Its relevance to the induction of a protective antibody response is, however, still a matter of debate. We addressed this issue by using subviral dense bodies (DBs) of HCMV. DBs are exceptionally immunogenic. Laboratory HCMV strain DBs harbor important neutralizing antibody targets, like the glycoproteins B, H, L, M, and N, but they are devoid of the PC. To be able to directly compare the impact of the PC on the levels of neutralizing antibody (NT-abs) responses, a PC-positive variant of the HCMV laboratory strain Towne was established by bacterial artificial chromosome (BAC) mutagenesis (Towne-UL130rep). This strain synthesized PC-positive DBs upon infection of fibroblasts. These DBs were used in side-by-side immunizations with PC-negative Towne DBs. Mouse and rabbit sera were tested to address the impact of the PC on DB immunogenicity. The neutralizing antibody response to PC-positive DBs was superior to that of PC-negative DBs, as tested on fibroblasts, epithelial cells, and endothelial cells and for both animal species used. The experiments revealed the potential of the PC to enhance the antibody response against HCMV. Of particular interest was the finding that PC-positive DBs induced an antibody response that blocked the infection of fibroblasts by a PC-positive viral strain more efficiently than sera following immunizations with PC-negative particles.IMPORTANCE Infections with the human cytomegalovirus (HCMV) may cause severe and even life-threatening disease manifestations in newborns and immunosuppressed individuals. Several strategies for the development of a vaccine against this virus are currently pursued. A critical question in this respect refers to the antigenic composition of a successful vaccine. Using a subviral particle vaccine candidate, we show here that one protein complex of HCMV, termed the pentameric complex (PC), enhances the neutralizing antibody response against viral infection of different cell types. We further show for the first time that this not only relates to the infection of epithelial or endothelial cells; the presence of the PC in the particles also enhanced the neutralizing antibody response against the infection of fibroblasts by HCMV. Together, these findings argue in favor of including the PC in strategies for HCMV vaccine development.