Adeno-associated virus serotype 4 (AAV4) and AAV5 both require sialic acid binding for hemagglutination and efficient transduction but differ in sialic acid linkage specificity
Adeno-associated virus serotype 4 (AAV4) and AAV5 both require sialic acid binding for hemagglutination and efficient transduction but differ in sialic acid linkage specificity
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DOI：
10.1128/jvi.75.15.6884-6893.2001
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发表时间：
2001-08-01
影响因子：
5.4
通讯作者：
Chiorini, JA
中科院分区：
文献类型：
--
作者：
Kaludov, N;Brown, KE;Chiorini, JA
腺相关病毒血清型4（AAV 4）和AAV 5与AAV 2相比具有不同的嗜性并且彼此之间具有不同的嗜性。我们最近报道了a2 -3唾液酸是AAV 5结合和转导所必需的。在这项研究中，我们表征了AAV 4的结合和转导，发现它也结合唾液酸，但特异性与AAV 5显著不同。AAV 4可以凝集来自几个物种的红细胞，而AAV 5仅凝集恒河猴红细胞。用胰蛋白酶处理红细胞可抑制AAV 4和AAV 5的血细胞凝集，表明凝集素是一种蛋白质。用神经氨酸酶处理Cos和红细胞也表明AAV 4结合α 2 -3唾液酸。然而，用神经氨酸酶处理的红细胞的再唾液酸化实验证明，AAV 4结合需要α 2 -3 O-连接的唾液酸，而AAV 5需要N-连接的唾液酸。同样，唾液酸缺陷CHO细胞的再唾液酸化也支持这一结论。通过与糖基化的N-连接或O-连接抑制剂一起孵育的细胞的结合和转导实验证实了对AAV 4和AAV 5的连接特异性的差异。此外，AAV 4转导仅被可溶性α 2 -3唾液酸阻断，而AAV 5可以被α 2 -3或α 2 -6唾液酸阻断。这些结果表明，AAV 4和AAV 5需要不同的含唾液酸的糖蛋白来结合和转导靶细胞，并且它们进一步解释了AAV 4和AAV 5的不同向性。
Adeno-associated virus serotype 4 (AAV4) and AAV5 have different tropisms compared to AAV2 and to each other. We recently reported that alpha2-3 sialic acid is required for AAV5 binding and transduction. In this study, we characterized AAV4 binding and transduction and found it also binds sialic acid, but the specificity is significantly different from AAV5. AAV4 can hemagglutinate red blood cells from several species, whereas AAV5 hemagglutinates only rhesus monkey red blood cells. Treatment of red blood cells with trypsin inhibited hemagglutination for both AAV4 and AAV5, suggesting that the agglutinin is a protein. Treatment of Cos and red blood cells with neuraminidases also indicated that AAV4 bound alpha2-3 sialic acid. However, resialylation experiments with neuraminidase-treated red blood cells demonstrated that AAV4 binding required alpha2-3 O-linked sialic acid, whereas AAV5 required N-linked sialic acid. Similarly, resialylation of sialic acid-deficient CHO cells supported this same conclusion. The difference in linkage specificity for AAV4 and AAV5 was confirmed by binding and transduction experiments with cells incubated with either N-linked or O-linked inhibitors of glycosylation. Furthermore, AAV4 transduction was only blocked with soluble alpha2-3 sialic acid, whereas AAV5 could be blocked with either alpha2-3 or alpha2-6 sialic acid. These results suggest that AAV4 and AAV5 require different sialic acid-containing glycoproteins for binding and transduction of target cells and they further explain the different tropism of AAV4 and AAV5.