Detection of HIV-1 DNA in microglia/macrophages, astrocytes and neurons isolated from brain tissue with HIV-1 encephalitis by laser capture microdissection.
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.
Division of Molecular Virology, St. Luke's-Roosevelt Hospital Center, College of Physicians & Surgeons, Columbia University, New York, NY 10019, USA.
G
G
Trillo-Pazos
A
A
Diamanturos
L
L
Rislove
T
T
Menza
W
W
Chao
P
P
Belem
S
S
Sadiq
S
S
Morgello
L
L
Sharer
D J
DJ
Volsky
D000293:Adolescent; D000328:Adult; D000368:Aged; D001253:Astrocytes / Q000473:pathology / Q000821:virology; D001479:Basal Ganglia / Q000473:pathology / Q000821:virology*; D002648:Child; D004279:DNA, Viral / Q000032:analysis; D018792:Encephalitis, Viral / Q000473:pathology / Q000821:virology*; D005625:Frontal Lobe / Q000473:pathology / Q000821:virology*; D015683:Gene Products, gag / Q000235:genetics / Q000302:isolation & purification; D016655:HIV Core Protein p24 / Q000302:isolation & purification*; D015658:HIV Infections / Q000473:pathology* / Q000821:virology; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification; D006801:Humans; D008264:Macrophages / Q000473:pathology / Q000821:virology; D017628:Microglia / Q000473:pathology / Q000821:virology; D008846:Micromanipulation / Q000379:methods; D018613:Microscopy, Confocal; D008875:Middle Aged; D009474:Neurons / Q000473:pathology / Q000821:virology
D004279:DNA, Viral; D015683:Gene Products, gag; D016655:HIV Core Protein p24