Successful isolation of infectious and high titer human monocyte-derived HIV-1 from two subjects with discontinued therapy.
Successful isolation of infectious and high titer human monocyte-derived HIV-1 from two subjects with discontinued therapy.
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从两名停止治疗的受试者中成功分离出传染性高滴度人类单核细胞来源的 HIV-1
DOI：
10.1371/journal.pone.0065071
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Zhu T
中科院分区：
文献类型：
--
作者：
Wang T;Xu Y;Zhu H;Andrus T;Ivanov SB;Pan C;Dolores J;Dann GC;Zhou M;Forte D;Yang Z;Holte S;Corey L;Zhu T
血液单核细胞中的HIV-1 DNA被认为是各种HIV-1感染组织巨噬细胞的病毒来源，这也被称为“特洛伊木马”假说。然而，这些DNA是否可以产生病毒体多年来一直是一个悬而未决的问题，因为无法直接从单核细胞中分离高滴度和感染性的HIV-1。在本研究中，我们证明了从四名研究受试者中的两名中成功分离出两种M-HIV-1毒株（1690 M和1175 M），以及其体内对照，从CD 4 + T细胞中分离的HIV-1（T-HIV-1），1690 T和1175 T。所有M-和T-HIV-1分离株均检测到CCR 5嗜性。两种M-HIV-1在单核细胞衍生的巨噬细胞（MDM）中表现出比两种T-HIV-1更高的复制水平。与我们之前关于受试者1175晚期感染的报告一致，在1175 M和1175 T之间鉴定了区室化的env C2-V3-C3序列。相比之下，从感染相对较早的受试者1690中分离的1690 M和1690 T显示出同源的env C2-V3-C3序列。然而，在1690 M的Gag-Pol区域中检测到多种逆转录酶（RT）抑制剂耐药相关变异，但在1690 T中未检测到。通过进一步测量MDM感染后的HIV DNA细胞内拷贝数，发现1690 M在巨噬细胞中具有比1690 T显著更高的DNA合成效率，表明更高的RT活性，这通过AZT抑制试验证实。这些结果表明，M-和T-HIV-1分别在两个研究对象中被区室化。因此，我们证明，在体外条件下，HIV-1感染的人单核细胞可以有效地释放活病毒，同时分化为巨噬细胞。
HIV-1 DNA in blood monocytes is considered a viral source of various HIV-1 infected tissue macrophages, which is also known as “Trojan horse” hypothesis. However, whether these DNA can produce virions has been an open question for years, due to the inability of isolating high titer and infectious HIV-1 directly from monocytes. In this study, we demonstrated successful isolation of two strains of M-HIV-1 (1690 M and 1175 M) from two out of four study subjects, together with their in vivo controls, HIV-1 isolated from CD4+ T-cells (T-HIV-1), 1690 T and 1175 T. All M- and T- HIV-1 isolates were detected CCR5-tropic. Both M- HIV-1 exhibited higher levels of replication in monocyte-derived macrophages (MDM) than the two T- HIV-1. Consistent with our previous reports on the subject 1175 with late infection, compartmentalized env C2-V3-C3 sequences were identified between 1175 M and 1175 T. In contrast, 1690 M and 1690 T, which were isolated from subject 1690 with relatively earlier infection, showed homogenous env C2-V3-C3 sequences. However, multiple reverse transcriptase (RT) inhibitor resistance-associated variations were detected in the Gag-Pol region of 1690 M, but not of 1690 T. By further measuring HIV DNA intracellular copy numbers post-MDM infection, 1690 M was found to have significantly higher DNA synthesis efficiency than 1690 T in macrophages, indicating a higher RT activity, which was confirmed by AZT inhibitory assays. These results suggested that the M- and T- HIV-1 are compartmentalized in the two study subjects, respectively. Therefore, we demonstrated that under in vitro conditions, HIV-1 infected human monocytes can productively release live viruses while differentiating into macrophages.
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影响因子：
3.3
作者：
Bergamaschi A;Pancino G
通讯作者：
Pancino G
DOI：
10.1073/pnas.94.24.13193
发表时间：
1997-11-25
影响因子：
11.1
作者：
Chun, TW;Stuyver, L;Fauci, AS
通讯作者：
Fauci, AS
影响因子：
64.8
作者：
EMBRETSON, J;ZUPANCIC, M;HAASE, AT
通讯作者：
HAASE, AT
影响因子：
7.3
作者：
Cywin, CL;Klunder, JM;Hattox, SE
通讯作者：
Hattox, SE
影响因子：
5.4
作者：
Arfi, Vanessa;Riviere, Lise;Cimarelli, Andrea
通讯作者：
Cimarelli, Andrea