Division of Retrovirology, United States Military HIV Research Program, Walter Reed Army Institute of Research, Rockville, Maryland, United States of America.
PLoS One. 2009 Dec 14;4(12):e8297. doi: 10.1371/journal.pone.0008297.
Although CD4(+) cells represent the major target for HIV infection in blood, claims of complement-independent binding of HIV-1 to erythrocytes and the possible role of Duffy blood group antigen, have generated controversy. To examine the question of binding to erythrocytes, HIV-1 was incubated in vitro with erythrocytes from 30 healthy leukapheresis donors, and binding was determined by p24 analysis and adsorption of HIV-1 with reduction of infectivity for CD4(+) target cells. All of the cells, regardless of blood group type, bound HIV-1 p24. A typical preparation of erythrocytes bound <2.4% of the added p24, but erythrocytes selectively removed essentially all of the viral infectivity as determined by decreased infection of CD4(+) target cells; however, cell-associated HIV-1 was approximately 100-fold more efficient, via trans infection, than unadsorbed virus for infection of CD4(+) cells. All of the bound HIV-1 p24 was released by treatment of the cells with EDTA, and binding was optimized by adding Ca(2+) and Mg(2+) during the washing of erythrocytes containing bound HIV-1. Although the small number of contaminating leukocytes in the erythrocyte preparation also bound HIV-1 p24, there was no significant binding to CD4, and it thus appears that the binding occurred on leukocytes at non-CD4 sites. Furthermore, binding occurred to erythrocyte ghosts from which contaminating leukocytes had been previously removed. The results demonstrate that erythrocytes incubated in vitro with HIV-1 differentially adsorb all of the infectious HIV-1 virions (as opposed to non-infectious or degraded virions) in the absence of complement and independent of blood group, and binding is dependent on divalent cations. By analogy with HIV-1 bound to DC-SIGN on dendritic cells, erythrocyte-bound HIV-1 might comprise an important surface reservoir for trans infection of permissive cells.
虽然 CD4(+)细胞是 HIV 感染的主要靶标,但 HIV-1 与红细胞的补体非依赖性结合以及 Duffy 血型抗原的可能作用引起了争议。为了研究与红细胞结合的问题,将 HIV-1 在体外与 30 位健康白细胞单采供体的红细胞孵育,并通过 p24 分析和吸附 HIV-1 以及降低对 CD4(+)靶细胞的感染性来确定结合。所有的细胞,无论血型如何,都结合了 HIV-1 p24。典型的红细胞制剂结合了<2.4%的添加 p24,但红细胞选择性地去除了几乎所有的病毒感染力,通过降低 CD4(+)靶细胞的感染来确定;然而,与未吸附的病毒相比,通过转染,细胞相关的 HIV-1 对于感染 CD4(+)细胞的效率约高 100 倍。用 EDTA 处理细胞可释放所有结合的 HIV-1 p24,并且通过在含有结合的 HIV-1 的红细胞洗涤过程中添加 Ca(2+)和 Mg(2+)来优化结合。尽管红细胞制剂中少量的污染白细胞也结合了 HIV-1 p24,但它们没有与 CD4 显著结合,因此似乎结合发生在非 CD4 位点的白细胞上。此外,结合发生在先前去除了污染白细胞的红细胞胞。结果表明,在不存在补体的情况下,体外与 HIV-1 孵育的红细胞以非依赖性方式差异吸附所有传染性 HIV-1 病毒颗粒(而不是非感染性或降解的病毒颗粒),并且结合依赖于二价阳离子。通过与树突状细胞上的 DC-SIGN 结合的 HIV-1 相类比,红细胞结合的 HIV-1 可能构成了允许细胞转染的重要表面储库。