title: Impaired accessory cell function in a human dendritic cell line after human immunodeficiency virus infection.
abstract: We generated human dendritic cell (DC) hybridoma cell lines by fusing HGPRT-deficient promonocytic U937 cells with immature DCs obtained by culturing peripheral blood monocytes with interleukin-4 (IL-4; 1,000 U/ml) and granulocyte-macrophage colony-stimulating factor (100 U/ml) for 7 days and mature DCs by treatment with tumor necrosis factor alpha (12.5 microg/ml) for 3 days. Only one fusion with immature DCs was successful and yielded four cell lines--HB-1, HB-2, HB-3, and HB-9--with an overall fusion efficiency of 0.0015%. The cell lines were stable in long-term culture, displayed morphological features typical of DCs, and expressed distinct class I and class II molecules not present on U937 (A*031012, B*51011, Cw*0701, DRB3*01011 52, and DR5*01011). A representative cell line, HB-2, that expressed DC markers including CD83, CD80 and CD86 could be induced to produce IL-12 through CD40 stimulation. After human immunodeficiency virus (HIV) infection, there was impairment of antigen-presenting cell (APC) function, which was manifested by an inability to stimulate allogeneic T-cell responses. There was no change in expression of major histocompatibility complex class I and class II antigens, CD83, CD40, CD4, CD11c, CD80, CD86, CD54, and CD58, or IL-12 production in the HIV-infected HB-2 cells. The HIV-infected HB-2 cells induced T-cell apoptosis in the cocultures. T-cell proliferation could be partially restored by using ddI, indinivir, and blocking anti-gp120 and anti-IL-10 antibodies. Our data suggest that there are multiple mechanisms that DCs use to inhibit T-cell responses in HIV-infected patients. The HB-2 cell line could be a useful model system to study APC function in HIV-infected DCs.
authors.affiliation: Division of Clinical Immunology, Box 1089, 1 Gustave Levy Place, Mount Sinai School of Medicine, New York, NY 10029, USA.
authors.forename: Prarthana
authors.initials: P
authors.lastname: Beuria
authors.forename: Houchu
authors.initials: H
authors.lastname: Chen
authors.forename: Michael
authors.initials: M
authors.lastname: Timoney
authors.forename: Kirk
authors.initials: K
authors.lastname: Sperber
mesh_terms: D017951:Antigen Presentation; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D018920:Coculture Techniques; D003713:Dendritic Cells / Q000276:immunology / Q000502:physiology / Q000821:virology*; D015658:HIV Infections / Q000276:immunology* / Q000473:pathology; D015395:Histocompatibility Antigens Class I / Q000032:analysis; D006801:Humans; D006825:Hybridomas; D007959:Lymphocyte Culture Test, Mixed; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000821:virology
chemical_list: D015395:Histocompatibility Antigens Class I