Large-Scale Screening of HCMV-Seropositive Blood Donors Indicates that HCMV Effectively Escapes from Antibodies by Cell-Associated Spread.
Large-Scale Screening of HCMV-Seropositive Blood Donors Indicates that HCMV Effectively Escapes from Antibodies by Cell-Associated Spread.
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DOI：
10.3390/v10090500
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发表时间：
2018-09-14
期刊：
影响因子：
--
通讯作者：
Sinzger C
中科院分区：
文献类型：
--
作者：
Falk JJ;Winkelmann M;Laib Sampaio K;Paal C;Schrezenmeier H;Alt M;Stanton R;Krawczyk A;Lotfi R;Sinzger C
免疫球蛋白对治疗人巨细胞病毒（HCMV）感染仅中度有效，可能是由于对细胞相关病毒传播无效。为了克服这一局限性，我们的目标是鉴定血浆中具有特殊抗体的个体，这些抗体可以有效地阻断HCMV的细胞相关传播。产生细胞相关HCMV毒株Merlin的分泌Gaussia β-内酰胺酶的突变体，并评价荧光素酶活性作为细胞相关病灶扩散程度的读数。然后应用这种基于报告病毒的测定筛选来自8400个HCMV血清阳性个体的血浆样品的抑制作用，包括直接作用的抗病毒药物作为阳性对照。所有血浆均未将病毒传播降低至这些对照品的水平。即使是在筛选试验中部分降低荧光素酶活性的得分最高的样品，当用更准确的基于免疫荧光的试验重新评估时，也未能抑制病灶生长。选择的血清对游离病毒的中和能力高，分别进行了分析，他们都没有阻止最近的三个临床HCMV分离株的焦点传播，也没有减少粒子的数量传输，证明与荧光梅林突变体。我们的结论是，与细胞间传播抑制血浆的捐助者是不存在的或极其罕见的，强调细胞相关的传播作为一个非常有效的免疫逃逸机制的HCMV。
Immunoglobulins are only moderately effective for the treatment of human cytomegalovirus (HCMV) infections, possibly due to ineffectiveness against cell-associated virus spread. To overcome this limitation, we aimed to identify individuals with exceptional antibodies in their plasma that can efficiently block the cell-associated spread of HCMV. A Gaussia luciferase-secreting mutant of the cell-associated HCMV strain Merlin was generated, and luciferase activity evaluated as a readout for the extent of cell-associated focal spread. This reporter virus-based assay was then applied to screen plasma samples from 8400 HCMV-seropositive individuals for their inhibitory effect, including direct-acting antiviral drugs as positive controls. None of the plasmas reduced virus spread to the level of these controls. Even the top-scoring samples that partially reduced luciferase activity in the screening assay failed to inhibit focal growth when reevaluated with a more accurate, immunofluorescence-based assay. Selected sera with high neutralizing capacity against free viruses were analyzed separately, and none of them prevented the focal spread of three recent clinical HCMV isolates nor reduced the number of particles transmitted, as demonstrated with a fluorescent Merlin mutant. We concluded that donors with cell-to-cell-spread-inhibiting plasma are nonexistent or extremely rare, emphasizing cell-associated spread as a highly efficient immune escape mechanism of HCMV.
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DOI：
10.1099/vir.0.018994-0
发表时间：
2010-06
期刊：
The Journal of general virology
影响因子：
--
作者：
Dargan DJ;Douglas E;Cunningham C;Jamieson F;Stanton RJ;Baluchova K;McSharry BP;Tomasec P;Emery VC;Percivalle E;Sarasini A;Gerna G;Wilkinson GW;Davison AJ
通讯作者：
Davison AJ
影响因子：
3.8
作者：
Gerna, Giuseppe;Sarasini, Antonella;Revello, M. Grazia
通讯作者：
Revello, M. Grazia
影响因子：
6.4
作者：
Falk, Jessica Julia;Winkelmann, Martina;Sinzger, Christian
通讯作者：
Sinzger, Christian
影响因子：
4.9
作者：
Kauvar, Lawrence M.;Liu, Keyi;Pereira, Lenore
通讯作者：
Pereira, Lenore
影响因子：
11.8
作者：
Boppana, Suresh B.;Ross, Shannon A.;Fowler, Karen B.
通讯作者：
Fowler, Karen B.