Isolation of highly purified human blood monocytes for in vitro HIV-1 infection studies of monocyte/macrophages

Hassan, Nassef F.; Cutilli, Joann; Douglas, Steven D.

doi:10.1016/0022-1759(90)90058-4

Cited by 42 publications

(22 citation statements)

References 7 publications

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“…The blood samples were identified as HIV-1 antibody negative by anonymous testing with the ELISA method (Coulter). Monocytes were purified according to our previously described techniques (23,24).…”

Section: Methodsmentioning

confidence: 99%

Substance P antagonist (CP-96,345) inhibits HIV-1 replication in human mononuclear phagocytes

Lai

Zhan

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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Section: Methodsmentioning

confidence: 99%

Substance P antagonist (CP-96,345) inhibits HIV-1 replication in human mononuclear phagocytes

Lai

Zhan

et al. 2001

Proc. Natl. Acad. Sci. U.S.A.

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“…For infection with HIV-1, mononuclear cells were prepared from the peripheral blood of HIV-1 seronegative volunteers as described previously (22,24,25). After removal from the Ficoll-Hypaque interface, monocytes were further purified following the procedure described by Hassan et al (26). PBMC were suspended at 2-4 ϫ 10 6 /ml in DMEM (GIBCO-BRL) containing 20% horse serum (GIBCO-BRL) and 15 ml was aliquoted in 75-cm 2 tissue culture flasks (Corning Glass Works, Corning, NY) coated with 2% gelatin.…”

Section: Hiv-1mentioning

confidence: 99%

Modulation of the effector function of human macrophages for Histoplasma capsulatum by HIV-1. Role of the envelope glycoprotein gp120.

Chaturvedi

Newman²

1997

J. Clin. Invest.

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We have demonstrated that monocyte-derived macrophages (M ) from HIV ϩ individuals are deficient in their capacity to phagocytose Histoplasma capsulatum (Hc) yeasts, and are more permissive for the intracellular growth of Hc. To determine whether these defects in M function were caused by HIV infection of the M and/or by pathological events associated with HIV infection, cultured normal human M were infected with the HIV-1 BaL strain. Virus production, quantified by reverse transcriptase activity and p24 antigen, was evident on day 8 after infection and peaked on day 16. On days 12, 16, and 20 after infection, HIV-1-infected M were deficient in their capacity to recognize and bind Hc yeasts compared with control M , and also were more permissive for the intracellular growth of Hc. Culture of normal M with the envelope glycoprotein gp120 inhibited phagocytosis of Hc yeasts by M in a concentration-dependent manner, but did not cause more rapid intracellular growth of Hc. Normal M cultured in the serum of HIV ϩ individuals with impaired M function subsequently were deficient in their capacity to phagocytose Hc yeasts, and were more permissive for the intracellular growth of yeasts compared with M cultured in normal serum. Conversely, culture of normal M in the serum of HIV ϩ patients with normal M function did not affect the interaction of Hc yeasts with M . Moreover, when M from HIV ϩ individuals that were initially defective in host defense against Hc were cultured in normal HIV Ϫ serum, normal M function was demonstrated. Adsorption of gp120 from the serum of two HIV ϩ patients removed the capacity of the serum to cause a M defect in phagocytosis of Hc, but had no effect on the capacity of the serum to cause accelerated intracellular growth. These data demonstrate that observed defects in M interaction with Hc yeasts may be caused by gp120 and other, as yet unknown serum component(s) probably released into serum by HIV-infected cells. (

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“…Purified monocytes were detached by EDTA. After the initial purification, at least 97% of the cells were monocytes, as determined by nonspecific esterase staining and flow cytometry using MAb against CD14 and LDL specific for monocytes and macrophages as described previously (16,17). Cells were plated in 48-well culture plates at a density of 0.5 ϫ 10 6 cells/well in the DMEM containing 10% FCS to obtain freshly isolated monocytes (within 48 h of isolation), and monocyte-derived macrophage (7)(8) MIP-1␤ titration.…”

Section: Neonatal Monocyte Preparationmentioning

confidence: 99%

Morphine Enhances HIV Infection of Neonatal Macrophages

Merrill

Mooney

et al. 2003

Pediatr Res

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Isolation of highly purified human blood monocytes for in vitro HIV-1 infection studies of monocyte/macrophages

Cited by 42 publications

References 7 publications

Substance P antagonist (CP-96,345) inhibits HIV-1 replication in human mononuclear phagocytes

Substance P antagonist (CP-96,345) inhibits HIV-1 replication in human mononuclear phagocytes

Modulation of the effector function of human macrophages for Histoplasma capsulatum by HIV-1. Role of the envelope glycoprotein gp120.

Morphine Enhances HIV Infection of Neonatal Macrophages

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