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 LG 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 LG (10-25% of CD1a(+)/CD4(+) cells), deprived of contaminating T cells and then incubated with HIV-1(IIIB). After 24 hr, purified LC and LC-depleted EG fractions were obtained by immunomagnetic separation. Polymerase chain reaction (PGR) analysis showed the presence of HTV-1 proviral DNA (gag) only in purified LC. In addition, LC-enriched EC, purified LG, LC-depleted EG or the non-permissive cell line, TF-1, the latter having being previously challenged with HIV-1(IIIB) 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. Go-cultures of C8166 cells with purified LC or LC-enriched EG previously exposed to HIV-1(IIIB) 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.

In-Vitro Infection Of Human Epidermal Langerhans Cells With Hiv-1 / E., Ramazzotti; Marconi, Alessandra; M. C., Re; G., Girolomoni; G., Cenacchi; M., Vignoli; G., Zambruno; G., Furlini; M., La Placa; Giannetti, Alberto. - In: IMMUNOLOGY. - ISSN 0019-2805. - STAMPA. - 85:(1995), pp. 94-98.

In-Vitro Infection Of Human Epidermal Langerhans Cells With Hiv-1

MARCONI, Alessandra;GIANNETTI, Alberto
1995

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 LG 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 LG (10-25% of CD1a(+)/CD4(+) cells), deprived of contaminating T cells and then incubated with HIV-1(IIIB). After 24 hr, purified LC and LC-depleted EG fractions were obtained by immunomagnetic separation. Polymerase chain reaction (PGR) analysis showed the presence of HTV-1 proviral DNA (gag) only in purified LC. In addition, LC-enriched EC, purified LG, LC-depleted EG or the non-permissive cell line, TF-1, the latter having being previously challenged with HIV-1(IIIB) 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. Go-cultures of C8166 cells with purified LC or LC-enriched EG previously exposed to HIV-1(IIIB) 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.
85
94
98
In-Vitro Infection Of Human Epidermal Langerhans Cells With Hiv-1 / E., Ramazzotti; Marconi, Alessandra; M. C., Re; G., Girolomoni; G., Cenacchi; M., Vignoli; G., Zambruno; G., Furlini; M., La Placa; Giannetti, Alberto. - In: IMMUNOLOGY. - ISSN 0019-2805. - STAMPA. - 85:(1995), pp. 94-98.
E., Ramazzotti; Marconi, Alessandra; M. C., Re; G., Girolomoni; G., Cenacchi; M., Vignoli; G., Zambruno; G., Furlini; M., La Placa; Giannetti, Alberto
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Utilizza questo identificativo per citare o creare un link a questo documento: http://hdl.handle.net/11380/611877
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