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Detection of HIV-1 DNA in microglia/macrophages, astrocytes and neurons isolated from brain tissue with HIV-1
G Trillo-Pazos1, A Diamanturos, L Rislove
1Division of Molecular Virology, St. Luke's-Roosevelt Hospital Center, College of Physicians & Surgeons, Columbia University, New York, NY 10019, USA.
Abstract:
In HIV-1 encephalitis, HIV-1 replicates predominantly in macrophages and microglia. Astrocytes also carry HIV-1, but the infection of oligodendrocytes and neurons is debated. In this study we examined the presence of HIV-1 DNA in different brain cell types in 6 paraffin embedded, archival post-mortem pediatric and adult brain tissues with HIV-1 encephalitis by Laser Capture Microdissection (LCM). Sections from frontal cortex and basal ganglia were stained by immunohistochemistry for CD68 (microglia), GFAP (astrocytes), MAP2 (neurons), and p24 (HIV-1 positive cells) and different cell types were microdissected by LCM. Individual cells or pools of same type of cells were lysed, the cell lysates were subjected to PCR using HIV-1 gag SK38/SK39 primers, and presence of HIV-1 DNA was confirmed by Southern blotting. HIV-1 gag DNA was consistently detected by this procedure in the frontal cortex and basal ganglia in 1 to 20 p24 HIV-1 capsid positive cells, and in pools of 50 to 100 microglia/macrophage cells, 100 to 200 astrocytes, and 100 to 200 neurons in HIV-1 positive cases but not in HIV-1 negative controls. These findings suggest that in addition to microglia, the infection of astrocytes and neurons by HIV-1 may contribute to the development of HIV-1 disease in the brain.
Insights
Human Immunodeficiency Virus type 1 (HIV-1) infects brain cells in encephalitis. This study found HIV-1 DNA in astrocytes and neurons, suggesting they contribute to HIV-1 brain disease alongside microglia.
Area of Science:
- Neuroscience
- Virology
- Immunology
Background:
- Human Immunodeficiency Virus type 1 (HIV-1) encephalitis primarily involves macrophages and microglia.
- The role of HIV-1 infection in astrocytes, neurons, oligodendrocytes remains debated.
Purpose of the Study:
- To investigate the presence of HIV-1 DNA in distinct brain cell types in HIV-1 encephalitis.
- To determine if astrocytes and neurons are infected by HIV-1 in the brain.
Main Methods:
- Archival post-mortem pediatric and adult brain tissues with HIV-1 encephalitis were analyzed.
- Laser Capture Microdissection (LCM) was used to isolate specific cell types (microglia, astrocytes, neurons).
- HIV-1 DNA presence was confirmed using PCR and Southern blotting after cell lysis and staining for cell markers and HIV-1 p24.
Main Results:
- HIV-1 gag DNA was detected in microglia, astrocytes, and neurons from frontal cortex and basal ganglia.
- Infection was observed in individual HIV-1 p24 positive cells and pools of specific cell types.
- No HIV-1 DNA was found in control HIV-1 negative brain tissues.
Conclusions:
- HIV-1 infects astrocytes and neurons in the brain, not just microglia/macrophages.
- Astrocyte and neuron infection may play a role in the pathogenesis of HIV-1 brain disease.
- Findings highlight the broader cellular tropism of HIV-1 within the central nervous system.
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