title: Development and characterization of a human microglia cell model of HIV-1 infection.
abstract: 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.
authors.affiliation: Department of Pediatrics, Division of Infectious Diseases, University of California San Diego, La Jolla, CA, 92093-0672, USA.
authors.forename: Pratima
authors.initials: P
authors.lastname: Rawat
authors.affiliation: Department of Pediatrics, Division of Infectious Diseases, University of California San Diego, La Jolla, CA, 92093-0672, USA. saspector@ucsd.edu.|Rady Children's Hospital, San Diego, CA, 92123, USA. saspector@ucsd.edu.
authors.forename: Stephen A
authors.initials: SA
authors.lastname: Spector
mesh_terms: D015703:Antigens, CD / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation; D048430:Cell Shape; D018932:Chemokine CCL2 / Q000494:pharmacology; D015870:Gene Expression; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D016655:HIV Core Protein p24 / Q000235:genetics* / Q000378:metabolism; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D016173:Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D017628:Microglia / Q000187:drug effects / Q000473:pathology / Q000821:virology*; D008954:Models, Biological; D009000:Monocytes / Q000187:drug effects / Q000473:pathology / Q000821:virology*; D020932:Nerve Growth Factor / Q000494:pharmacology; D010587:Phagocytosis; D061251:Primary Cell Culture; D017382:Reactive Oxygen Species / Q000378:metabolism; D014779:Virus Replication
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; C111835:CCL2 protein, human; D018932:Chemokine CCL2; D016655:HIV Core Protein p24; D035683:MicroRNAs; D017382:Reactive Oxygen Species; D016173:Macrophage Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D020932:Nerve Growth Factor
keywords: HIV-1; HMG; Human microglia; MMG; Monocyte-derived microglia