title: In vitro infection of human epidermal Langerhans' cells with HIV-1.
abstract: Epidermal Langerhans' cells (LC) from human immunodeficiency virus type-1 (HIV-1)-infected patients harbour HIV-1 proviral DNA and RNA. In the present study, we investigated whether LC from epidermis of normal, HIV-seronegative subjects could be infected in vitro with HIV-1. Epidermal cells (EC) spontaneously detached from epidermal sheet cultures were enriched for LC (10-25% of CD1a+/CD4+ cells), deprived of contaminating T cells and then incubated with HIV-1IIIB. After 24 hr, purified LC and LC-depleted EC fractions were obtained by immunomagnetic separation. Polymerase chain reaction (PCR) analysis showed the presence of HIV-1 proviral DNA (gag) only in purified LC. In addition, LC-enriched EC, purified LC, LC-depleted EC or the non-permissive cell line, TF-1, the latter having being previously challenged with HIV-1IIIB for the same length of time as the EC, were co-cultivated with C8166 cells, and the co-cultures assessed for the presence of HIV DNA by PCR. Co-cultures of C8166 cells with purified LC or LC-enriched EC previously exposed to HIV-1IIIB exhibited a time-dependent increase in HIV proviral DNA. In contrast, PCR analysis of C8166 cells co-cultured with either LC-depleted EC or TF-1 cells gave negative results. Finally, C8166 cells co-cultured with HIV-infected LC formed syncytia, showed membrane budding and released numerous retroviral particles. The results indicate that LC from normal subjects can be infected in vitro with HIV and can transmit infection to myeloid cells. This in vitro model may help in understanding the regulation of HIV infection of LC.
authors.affiliation: Institute of Microbiology, University of Bologna, Italy.
authors.forename: E
authors.initials: E
authors.lastname: Ramazzotti
authors.forename: A
authors.initials: A
authors.lastname: Marconi
authors.forename: M C
authors.initials: MC
authors.lastname: Re
authors.forename: G
authors.initials: G
authors.lastname: Girolomoni
authors.forename: G
authors.initials: G
authors.lastname: Cenacchi
authors.forename: M
authors.initials: M
authors.lastname: Vignoli
authors.forename: G
authors.initials: G
authors.lastname: Zambruno
authors.forename: G
authors.initials: G
authors.lastname: Furlini
authors.forename: M
authors.initials: M
authors.lastname: La Placa
authors.forename: A
authors.initials: A
authors.lastname: Giannetti
mesh_terms: D002465:Cell Movement; D002478:Cells, Cultured; D004279:DNA, Viral / Q000032:analysis; D004817:Epidermis / Q000821:virology*; D015658:HIV Infections / Q000635:transmission / Q000821:virology*; D015497:HIV-1 / Q000302:isolation & purification*; D006801:Humans; D007801:Langerhans Cells / Q000821:virology*; D016133:Polymerase Chain Reaction; D011533:Proviruses / Q000302:isolation & purification
chemical_list: D004279:DNA, Viral