Detection of HIV-specific DNA sequences in epidermal Langerhans cells infected in vitro by means of a cell-free system

1432-069X

Epidermal Langerhans cell ; Human immunodeficiency virus (HIV) ; In vitro cell-free HIV infection

Springer Online Journal Archives 1860-2000

Medicine

Abstract As dendritic antigen-presenting cells in skin and mucous membranes, Langerhans cells (LC) are found in areas at risk of inoculation by the human immunodeficiency virus (HIV). LC have been reported as targets for HIV-1. The aim of the present study was to investigate whether LC can be experimentally infected by HIV provided by a cell-free infection system. A cell-free suspensions was prepared from viral particles provided by chronically infected cell lines (U937 or H9 cells) by low-speed centrifugation followed by 0.45-Μm filtration. LC-enriched epidermal cell (EC) suspensions with no CD3+ cells (assessed by flow cytometry and electron microscopy) and uninfected U937 cells (cell-free infection system control) were infected with two isolates (HTLVIII-B and RF). The infectiousness of the cell-free virus fluids was controlled on U937 cells where proviral DNA was amplified (gag, pol, and env gene sequences by the polymerase chain reaction, PCR) and release of virus particles into the supernatant was controlled either by measure of the reverse transcriptase (RT) activity or detection of viral RNA amplified by RT-PCR for the gag gene sequences). Proviral DNA (gag gene sequences) was found in LC-enriched epidermal cellular DNA from day 4 post-infection with isolate HTLVIII-B and from day 7 with isolate RF. Although the RT activity did not reach a significantly high level, viral RNA was found in the supernatant of LC-enriched EC cultures at the same time as proviral DNA was detected in LC. The findings indicate that the cell-free virus-infection model leads to a later-occurring infection and a lower HIV expression as compared with cell-to-cell transmission. This model allowed us to confirm the CD4-independent infection of isolated trypsin-treated LC and thus may be attractive for in vitro study of this type of infection.

Electronic Resource