Development and characterization of a human microglia cell model of HIV-1 infection.
Development and characterization of a human microglia cell model of HIV-1 infection.
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DOI：
10.1007/s13365-016-0472-1
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发表时间：
2017-03
影响因子：
3.2
通讯作者：
Spector SA
中科院分区：
文献类型：
--
作者：
Rawat P;Spector SA
小胶质细胞是中枢神经系统内HIV-1 （HIV）的主要储存库。然而，目前使用转化细胞系的模型不能代表原发小胶质细胞，并且越来越难以获得用于分离原发人小胶质细胞（HMG）的胎脑样本。在这里，我们描述了HIV感染的单核细胞来源的小胶质细胞（MMG）细胞模型，它概括了原发性HMG的感染。从健康供体分离的CD14+细胞与M-CSF、β -神经生长因子、GM-CSF和CCL2一起培养，并与HMG进行比较。MMG和HMG细胞感染HIV，用p24抗原检测病毒复制。MMG和HMG细胞在培养第2周时均呈梭形，末端几乎没有分支或未分支的突起，均为CD11b+/CD11c+/CD14+/CD45+/CD195+/HLADRlow/CD86low/CD80+。ht - h - μ胶质细胞和HMC3转化细胞系缺乏人类小胶质细胞特征基因（C1Q、GAS6、GPR34、MERTK、PROS1和P2RY12），而MMG细胞表达所有这些基因。此外，MMG表达了所有小胶质细胞特征miRNA （miR-99a， miR125b-5p和miR-342-3p）。在PMA刺激下，MMG和HMG均产生ROS并吞噬标记的酶酶颗粒。感染HIV的MMG和HMG在感染后30天的培养上清液中产生相同水平的HIV p24抗原。因此，我们开发了一种源自原代单核细胞的HIV感染小胶质细胞模型，该模型概括了HMG的表型和分子特性，优于转化细胞系，并且具有与HMG相似的HIV复制动力学。
Microglia cells are the major reservoir of HIV-1 (HIV) within the CNS. However, current models using transformed cell lines are not representative of primary microglia and fetal brain samples for isolation of primary human microglia (HMG) are increasingly difficult to obtain. Here, we describe a monocyte-derived microglia (MMG) cell model of HIV infection that recapitulates infection of primary HMG. CD14+ cells isolated from healthy donors were cultured with M-CSF, beta-nerve growth factor, GM-CSF, and CCL2, and compared to HMG. MMG and HMG cells were infected with HIV and viral replication was detected by p24 antigen. Both MMG and HMG cells were found to acquire spindle shape with few branched or unbranched processes at their ends during the second week in culture and both were found to be CD11b+/CD11c+/CD14+/CD45+/CD195+/HLADRlow/CD86low/CD80+. Whereas hT-Hμglia and HMC3 transformed cell lines are deficient in human microglia signature genes (C1Q, GAS6, GPR34, MERTK, PROS1, and P2RY12), MMG cells expressed all of these genes. Additionally, MMG expressed all the microglia signature miRNA (miR-99a, miR125b-5p, and miR-342-3p). Both MMG and HMG produced ROS and phagocytosed labeled zymosan particles upon PMA stimulation. MMG and HMG infected with HIV produced equivalent levels of HIV p24 antigen in culture supernatants for 30 days post-infection. Thus, we have developed and characterized a microglia cell model of HIV infection derived from primary monocytes that recapitulates the phenotypic and molecular properties of HMG, is superior to transformed cell lines, and has similar HIV replication kinetics to HMG.
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DOI：
10.1006/bbrc.1996.1112
发表时间：
1996-07-25
影响因子：
3.1
作者：
Imai, Y;Ibata, I;Kohsaka, S
通讯作者：
Kohsaka, S
影响因子：
6.2
作者：
Imai, Y;Kohsaka, S
通讯作者：
Kohsaka, S
影响因子：
3.4
作者：
Bassett, Tyler;Bach, Paxton;Chan, Hing Man
通讯作者：
Chan, Hing Man
影响因子：
4.8
作者：
Campbell, Grant R.;Spector, Stephen A.
通讯作者：
Spector, Stephen A.
影响因子：
3.8
作者：
Bagasra, O;Lavi, E;Pomerantz, RJ
通讯作者：
Pomerantz, RJ