title: Restricted HIV type 1 replication under serum-free culture conditions in human monocyte-derived macrophages.
abstract: Monocytes (MOs) and macrophages (MACs) are well-known targets for HIV-1 infection. Even though the virus load is contributed mainly to lymphocytes during the asymptomatic phase of infection, the expression of HIV-1 in MO/MACs seems to be important for the course of the disease. To establish a model for restricted HIV-1 expression in MACs in vitro, we cultured MO-derived MACs under different culture conditions and analyzed their susceptibility to HIV-1 infection as well as their capacity for virus replication in vitro. MACs cultured under serum-free conditions with M-CSF (M-MACs) remain viable and functionally active as assessed by the analysis of cytokine production. In addition, the levels of CD4, CD14, CCR5, and HLA-DR expression are comparable to those of serum-derived MACs (SER-MACs). However, serum-free MACs were less susceptible to HIV-1 infection, with only 9.5+/-4.5% (mean+/-SEM) of all cells being p24 antigen positive on day 22 as compared with 51+/-9% under serum conditions (p < 0.005). Reverse transcriptase (RT) activity in the culture supernatant of M-MACs was always about 100-fold lower than that of SER-MACs even when comparable amounts of cells were infected. The addition of serum to serum-free cultures increased the percentage of HIV-1 p24 antigen-positive cells (21+/-8% positive cells on day 22) and increased the RT activity, indicating that serum factors could be important for HIV-1 replication in MACs. Therefore we also switched SER-MACs to serum-free culture conditions and found a sharp decrease in RT activity. However, the RT level could always be rescued by the addition of serum, even after a long serum-free culture period. This effect was dependent on the serum concentration added, with as little as 0.1% serum being effective in reestablishing viral production as measured by RT activity. In conclusion, we show that serum has an important role in the replication of HIV-1 in MACs. Our results suggest that besides the role of CD4 and CCR5 other microenvironmental factors, e.g., growth factors, cytokines, or hormones, which are not provided by the target cell itself, are involved in the regulation of MAC infection and of replication by HIV-1.
authors.affiliation: Department of Hematology and Oncology, University of Regensburg, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Kreutz
authors.forename: V
authors.initials: V
authors.lastname: Eisert
authors.forename: H
authors.initials: H
authors.lastname: Rübsamen-Waigmann
authors.forename: R
authors.initials: R
authors.lastname: Andreesen
authors.forename: H
authors.initials: H
authors.lastname: von Briesen
mesh_terms: D018929:Cell Culture Techniques; D016895:Culture Media, Serum-Free; D016655:HIV Core Protein p24 / Q000096:biosynthesis; D054303:HIV Reverse Transcriptase / Q000378:metabolism; D015497:HIV-1 / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D008264:Macrophages / Q000821:virology*; D009000:Monocytes / Q000821:virology*; D014779:Virus Replication
chemical_list: D016895:Culture Media, Serum-Free; D016655:HIV Core Protein p24; D054303:HIV Reverse Transcriptase