Detection of HIV-1 gene sequences in hippocampal neurons isolated from postmortem AIDS brains by laser capture microdissection.
We employed laser capture microdissection to remove individual pyramidal neurons from the CA1, CA3, and CA4 regions of formalin-fixed, paraffin-embedded hippocampus from 8 AIDS brains and 2 HIV-1-seronegative normal brains. We amplified HIV-1 gag and nef gene sequences using separate, double round PCR reactions for each of the primer sets. In all 3 hippocampal regions, amplification efficiency was best with sequence length between 284 and 324 bp; HIV-1 nef gene sequences were more common than HIV-1 gag sequences; and rank order for percent positive amplification was CA3 > CA4 > CA1 samples. These results are the first to detect HIV-1 gene sequences in microdissected human tissue. They indicate that brain neurons in vivo contain HIV-1 DNA sequences consistent with latent infection by this virus, and suggest that neurons display a selective vulnerability for HIV infection. Neuronal HIV infection could contribute to neuronal injury and death or act as a potential viral reservoir if reactivated.
Torres-Muñoz
J
J
Department of Pathology, University of Miami School of Medicine, Florida 33136, USA.
Stockton
P
P
Tacoronte
N
N
Roberts
B
B
Maronpot
R R
RR
Petito
C K
CK
D015526:AIDS Dementia Complex / Q000473:pathology* / Q000821:virology*; D000328:Adult; D001253:Astrocytes / Q000473:pathology / Q000821:virology; D002648:Child; D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis; D005260:Female; D015683:Gene Products, gag / Q000235:genetics; D015693:Gene Products, nef / Q000235:genetics; D015497:HIV-1 / Q000235:genetics / Q000302:isolation & purification*; D006624:Hippocampus / Q000473:pathology* / Q000821:virology; D006801:Humans; D007223:Infant; D007834:Lasers; D008297:Male; D009474:Neurons / Q000473:pathology* / Q000821:virology; D016133:Polymerase Chain Reaction; D054311:nef Gene Products, Human Immunodeficiency Virus
D017931:DNA Primers; D004279:DNA, Viral; D015683:Gene Products, gag; D015693:Gene Products, nef; D054311:nef Gene Products, Human Immunodeficiency Virus