Reversible Human Immunodeficiency Virus Type-1 Latency in Primary Human Monocyte-Derived Macrophages Induced by Sustained M1 Polarization
Reversible Human Immunodeficiency Virus Type-1 Latency in Primary Human Monocyte-Derived Macrophages Induced by Sustained M1 Polarization
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DOI：
10.1038/s41598-018-32451-w
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发表时间：
2018-09-24
影响因子：
4.6
通讯作者：
Poli, Guido
中科院分区：
文献类型：
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作者：
Graziano, Francesca;Aimola, Giulia;Poli, Guido
我们已经报道了用干扰素- γ (ifn - γ)和肿瘤坏死因子- α (tnf - α)短期刺激原代人单核细胞源性巨噬细胞（MDM），即M1极化，可以显著遏制病毒复制。在这里，我们发现在感染后7天用这些细胞因子重新刺激M1-MDM （M1(2) MDM）促进了HIV-1复制的限制增加，其特征是病毒产生水平非常低，接近检测不到的水平。与对照组和未重新刺激的M1-MDM相比，M1(2) MDM显示出更强的总HIV DNA和整合HIV DNA以及病毒mRNA表达的减少。M1(2) MDM的特点是在逆转录（RT）水平上上调限制性因子的表达，包括载脂蛋白B mRNA编辑酶、催化多肽样3A （APOBEC3A）和APOBEC3G，但不包括SAM结构域和含HD结构域的蛋白1（SAMHD1）。M1(2) MDM还显示了II类转激活子（CIITA）和Tripartite Motif22 （TRIM22）的表达增加，这两种前病毒转录的负调节因子，而HIV-1转录诱导剂的表达和磷酸化，如核因子kB （NF-kB）和转录信号传导和激活子1(STAT1)，在这些细胞中没有受损。M1(2) MDM感染的几乎静止状态通过与丝裂原刺激的白细胞共培养或与滤过的培养上清细胞孵育迅速逆转。M1(2) MDM含有复制能力强的HIV-1，因为RT抑制剂拉米夫定/3TC完全阻止了细胞刺激后病毒的传播。在感染单圆形水泡性口炎病毒- g伪型HIV-1的细胞中，M1(2) MDM中的前病毒表达被选择性地重新激活，而在对照组或未被重新激活的M1-MDM中则没有。因此，M1(2) MDM代表了一种可逆的、几乎静止的人原代巨噬细胞HIV-1感染的体外模型，可以进一步用于“治愈”相关研究。
We have reported that short-term stimulation of primary human monocyte-derived macrophages (MDM) with interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha), i.e. M1 polarization, leads to a significant containment of virus replication. Here we show that M1-MDM restimulation with these cytokines 7 days after infection (M1(2) MDM) promoted an increased restriction of HIV-1 replication characterized by very low levels of virus production near to undetectable levels. In comparison to control and M1-MDM that were not restimulated, M1(2) MDM showed a stronger reduction of both total and integrated HIV DNA as well as of viral mRNA expression. M1(2) MDM were characterized by an upregulated expression of restriction factors acting at the level of reverse transcription (RT), including apolipoprotein B mRNA editing enzyme, catalytic polypeptide-like 3A (APOBEC3A) and APOBEC3G, but not SAM domain and HD domain-containing protein 1 (SAMHD1). M1(2) MDM also showed an increased expression of Class II Transactivator (CIITA) and Tripartite Motif22 (TRIM22), two negative regulators of proviral transcription, whereas expression and phosphorylation of transcriptional inducers of HIV-1, such as nuclear factor kB (NF-kB) and signal transducer and activator of transcription 1 (STAT1), were not impaired in these cells. The almost quiescent state of the infection in M1(2) MDM was promptly reversed by coculture with mitogen-stimulated leukocytes or cell incubation with their filtered culture supernatant. M1(2) MDM harbored replication-competent HIV-1 as virus spreading following cell stimulation was fully prevented by the RT inhibitor lamivudine/3TC. Selective reactivation of proviral expression in M1(2) MDM, but not in control or in M1-MDM that were not restimulated, was confirmed in cells infected with single round Vesicular Stomatitis Virus-G-pseudotyped HIV-1. Thus, M1(2) MDM represent an in vitro model of reversible, almost quiescent HIV-1 infection of primary human macrophages that could be further exploited for "Cure" related investigations.