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HIV‐1 viral cDNA detection in human microdissected single neurons from AIDS with encephalitis autopsy cases
Author(s) -
TorresMunoz Jorge Enrique,
Nunez Mariana,
Petito Carol K.
Publication year - 2007
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.21.5.a404-b
Subject(s) - complementary dna , cd8 , virology , biology , cytotoxic t cell , virus , neuron , encephalitis , microbiology and biotechnology , immunology , gene , genetics , immune system , neuroscience , in vitro
We hypothesized that the previously‐demonstrated perineuronal CD8+ granzyme B+ cytotoxic T‐cells hone to HIV‐1‐infected neurons. We used laser capture microscope to remove neurons with and without adjacent CD8+ T‐lymphocytes from post‐mortem hippocampus of brains with HIV‐1 encephalitis and HIV‐1‐negative controls; their average post‐mortem intervals were 23 h and 17 h. We extracted DNA, submitted it to isothermal whole genome amplification and performed two rounds of real time PCR for sequence amplification. GA3PDH sequences were found in 87% of samples with 15–30 neurons; 33% of samples with 3–10 or 1 neuron and 50% of single neurons from controls. In the subset of GA3PDH‐positive samples, we found HIV‐1 gag, pol, or nef sequences in 38%, 33% and 33% of samples of 15–30, 3–10 and 1 neuron; viral sequences were absent in controls. HIV‐1 viral cDNA was present in 40% of CD8+ neuronal samples versus 12.5% of CD8‐ samples. Our results confirm prior studies by ourselves and others showing neuronal infection by HIV‐1 and show that HIV‐1 viral cDNA can be amplified from single neurons. The higher incidence of viral sequences in the CD8+ neuronal samples supports our hypothesis that CD8 T‐cells specifically hone to infected neurons in AIDS brains where they may participate in neuronal injury and death. Supported in part by NS RO1‐31977; controls from NICHD Brain and Tissue Bank (NO1‐HD‐4‐3383 and NO1‐HD‐4‐3368).