title: Astrocyte differentiation selectively upregulates CCL2/monocyte chemoattractant protein-1 in cultured human brain-derived progenitor cells.
abstract: Chemokines (chemoattractant cytokines) and their receptors are present in the brain and may play roles in both neurodevelopment and neuropathology. Increased brain levels of monocyte chemoattractant protein-1 (MCP-1), also known as CCL2, are found in patients with human immunodeficiency virus type 1 (HIV-1)-associated dementia and other acute and chronic neurologic diseases. Although the function of CCL2 in the brain is unclear, it is believed that upregulation of this chemokine during neuropathologic or neuroinflammatory conditions leads to recruitment of activated monocytes into the brain, where they differentiate into macrophages producing neurotoxic and inflammatory molecules. We recently showed that human fetal brain-derived progenitor cells are susceptible to HIV-1 and JC virus infection, and that differentiation toward an astrocyte phenotype increased virus production from these cells. In the current study, we found that in the absence of infection, progenitors produced moderate levels of CCL2 (5.6 ng per million cells). Astrocyte differentiation over 3 weeks increased CCL2 protein levels 30-fold in a biphasic manner, whereas neuronal differentiation decreased production 20-fold. Electromobility shift assays (EMSAs) demonstrated increased nuclear NF-kappaB levels within 2 h of initiating astrocyte differentiation, and inhibitors of NF-kappaB activation partially blocked the CCL2 increase in differentiating astrocytes. Transfection of progenitors with mutated CCL2 promoter/CAT reporter constructs showed that the distal promoter region, containing NF-kappaB and NF-I binding sites, is important for differentiation-induced CCL2 upregulation. Together these results suggest that the transcription factor NF-kappaB, and possibly NF-I, contribute to the upregulation of CCL2 chemokine production during the differentiation of human progenitor cells toward an astrocyte phenotype.
authors.affiliation: Laboratory of Molecular Medicine and Neuroscience, National Institute of Neurologic Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Diane M P
authors.initials: DM
authors.lastname: Lawrence
authors.forename: Pankaj
authors.initials: P
authors.lastname: Seth
authors.forename: Linda
authors.initials: L
authors.lastname: Durham
authors.forename: Frank
authors.initials: F
authors.lastname: Diaz
authors.forename: Roosevelt
authors.initials: R
authors.lastname: Boursiquot
authors.forename: Richard M
authors.initials: RM
authors.lastname: Ransohoff
authors.forename: Eugene O
authors.initials: EO
authors.lastname: Major
mesh_terms: D015526:AIDS Dementia Complex / Q000276:immunology / Q000378:metabolism / Q000503:physiopathology; D001253:Astrocytes / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D001665:Binding Sites / Q000502:physiology; D002454:Cell Differentiation / Q000235:genetics / Q000276:immunology*; D002478:Cells, Cultured; D002490:Central Nervous System / Q000166:cytology / Q000196:embryology* / Q000378:metabolism; D018932:Chemokine CCL2 / Q000235:genetics / Q000378:metabolism*; D002633:Chemotaxis / Q000235:genetics / Q000276:immunology; D015497:HIV-1 / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007577:JC Virus / Q000276:immunology / Q000378:metabolism; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism; D051839:NFI Transcription Factors / Q000235:genetics / Q000378:metabolism; D010641:Phenotype; D027601:Polyomavirus Infections / Q000276:immunology / Q000378:metabolism / Q000503:physiopathology; D011401:Promoter Regions, Genetic / Q000235:genetics; D013234:Stem Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism*; D015533:Transcriptional Activation / Q000502:physiology; D014412:Tumor Virus Infections / Q000276:immunology / Q000378:metabolism / Q000503:physiopathology; D015854:Up-Regulation / Q000235:genetics / Q000276:immunology*
chemical_list: C111835:CCL2 protein, human; D018932:Chemokine CCL2; D016328:NF-kappa B; D051839:NFI Transcription Factors