Herpes simplex virus with highly reduced gD levels can efficiently enter and spread between human keratinocytes
Herpes simplex virus with highly reduced gD levels can efficiently enter and spread between human keratinocytes
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DOI：
10.1128/jvi.75.21.10309-10318.2001
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发表时间：
2001-11-01
影响因子：
5.4
通讯作者：
Johnson, DC
中科院分区：
文献类型：
--
作者：
Huber, MT;Wisner, TW;Johnson, DC
单纯疱疹病毒1型(HSV-1)在粘膜上皮和神经组织中的快速传播主要取决于病毒在极化细胞及其构成的组织中导航的能力。为了了解HSV的进入和病毒跨细胞连接的传播，我们之前已经鉴定了一种人类角质形成细胞系HaCaT。这些细胞似乎比许多用于繁殖HSV的培养细胞更能准确地反映体内感染的细胞。缺乏GE/GI的HSV突变体在上皮和神经元组织内的传播高度受损，在HaCaT细胞中也显示出细胞间传播的缺陷，但在其他未极化的细胞中则没有。单纯疱疹病毒gD通常被认为是进入细胞和在细胞间传播的绝对必需的，无论是在培养细胞中还是在体内。在这里，HSV-1 gD突变病毒F-US6kan被发现能有效地进入HaCaT细胞和正常人角质形成细胞，并可以在细胞之间传播，而不需要补充细胞提供gD。相比之下，进入和传播到其他细胞，特别是通常用于传播HSV的高度转化的细胞，效率极低。对F-US6kan的进一步分析表明，该突变体表达极低水平的GD(1/500野生型)。中和性抗GD单抗可抑制F-US6kan进入细胞，提示F-US6kan利用这一少量的GD进入细胞。HaCaT细胞高水平表达HSV gD受体HveC，并且针对HveC的抗体也可以抑制F-US6kan进入HaCaT细胞。有趣的是，抗HveC抗体并不能完全抑制野生型HSV-1进入HaCaT细胞。这些结果有助于揭示HSV和人类角质形成细胞的重要特性。HSV具有极低水平的关键受体结合糖蛋白，可以进入高水平受体表达的细胞。在这种情况下，过剩的gD可能有助于避免被抗gD抗体中和。
The rapid spread of herpes simplex virus type 1 (HSV-1) in mucosal epithelia and neuronal tissue depends primarily on the ability of the virus to navigate within polarized cells and the tissues they constitute. To understand HSV entry and the spread of virus across cell junctions, we have previously characterized a human keratinocyte cell line, HaCaT. These cells appear to reflect cells infected in vivo more accurately than many of the cultured cells used to propagate HSV. HSV mutants lacking gE/gI are highly compromised in spread within epithelial and neuronal tissues and also show defects in cell-to-cell spread in HaCaT cells, but not in other, nonpolarized cells. HSV gD is normally considered absolutely essential for entry and cell-to-cell spread, both in cultured cells and in vivo. Here, an HSV-1 gD mutant virus, F-US6kan, was found to efficiently enter HaCaT cells and normal human keratinocytes and could spread from cell to cell without gD provided by complementing cells. By contrast, entry and spread into other cells, especially highly transformed cells commonly used to propagate HSV, were extremely inefficient. Further analyses of F-US6kan indicated that this mutant expressed extraordinarily low (1/500 wild-type) levels of gD. Neutralizing anti-gD monoclonal antibodies inhibited entry of F-US6kan, suggesting F-US6kan utilized this small amount of gD to enter cells. HaCaT cells expressed high levels of an HSV gD receptor, HveC, and entry of F-US6kan into HaCaT cells could also be inhibited with antibodies specific for HveC. Interestingly, anti-HveC antibodies were not fully able to inhibit entry of wild-type HSV-1 into HaCaT cells. These results help to uncover important properties of HSV and human keratinocytes. HSV, with exceedingly low levels of a crucial receptor-binding glycoprotein, can enter cells expressing high levels of receptor. In this case, surplus gD may be useful to avoid neutralization by anti-gD antibodies.