Induction of IFN-alpha in peripheral blood mononuclear cells by HIV-infected monocytes. Restricted antiviral activity of the HIV-induced IFN.

Abstract

PBMC cocultured with HIV-infected monocytes for 12 to 48 h released high levels of IFN activity. IFN titers were directly dependent upon time after virus infection and level of HIV replication in infected cells. IFN induction in PBMC was evident with HIV-infected monocytes and PBMC and with myeloid and lymphoblastoid cell lines with at least three different HIV strains. In HIV-infected cell line pairs in which virus infection occurs in both productive and restricted forms, IFN induction in PBMC occurred only with productive infection. IFN activity was acid stable and completely neutralized by antibodies against IFN-alpha. Induction of IFN required cell-cell contact between HIV-infected cells and PBMC, but was independent of MHC compatibility. With PBMC co-cultured with autologous HIV-infected monocytes, IFN induction was highly selective: IL-1 beta, IL-6, or TNF-alpha activity and mRNA were not detected. Cell surface determinants on HIV-infected monocytes that induced IFN in PBMC remained active after fixation in 4% paraformaldehyde. Both adherent and nonadherent PBMC produced IFN after coculture with HIV-infected monocytes. Ability to produce IFN by PBMC was not affected by depletion of T cell, NK cell, B cell, or monocyte subpopulations. The IFN activity produced by PBMC cocultured with HIV-infected cells was about 20-fold less active than equal quantities of rIFN-alpha 2b for inhibition of HIV replication in monocytes and at low concentrations enhanced virus growth. Clinical studies with HIV-infected patients and parallel findings in animal lentivirus disease suggest an adverse role for IFN in disease progression. Conditions for induction of IFN in the culture system described in this report may mimic those in the HIV-infected patient. Defining the molecular basis for IFN induction, the cells that produce IFN, and the altered biologic activity of this important cytokine may provide insight into the pathogenesis of HIV disease.

Mechanisms of HIV-mediated blood-brain barrier compromise and leukocyte transmigration under the current antiretroviral era

“…A critical component of NeuroHIV, in addition to BBB integrity, is the transmigration of uninfected and HIV-infected leukocytes into the brain. 30 Peripheral blood mononuclear cells (PBMCs) were isolated from Leukopaks (healthy donors) and infected in vitro using the HIV ADA viral isolate 31 , 32 , 33 , 34 or molecular clone pNL(AD 8 ) (an R5-pseudotyped NL4-3 clone). 35 After about one cycle of replication, 18–24 h after infection, treatment with ATPγS began (3 μM or 15 μM).…”

“…Overall, our data indicates that healthy and HAD-associated eATP concentrations (3 μM and 15 μM eATP, respectively) only result in minimal alterations in the expression and surface localization of molecules involved in leukocyte transmigration, cell adhesion, and eATP metabolism in the presence and absence of HIV infection.

Figure 4 Focal adhesion molecules, junctional proteins, and eATP metabolism enzyme expression are not significantly altered after acute ATPγS treatment of HIV-infected or uninfected PBMCs PBMCs were isolated from Leukopaks derived from healthy patients, then infected in vitro with HIV ADA 31 , 32 , 33 , 34 or pNL(AD 8 ) (10 ng/mL). 35 PBMCs (infected and uninfected) were either untreated or treated with 3 μM or 15 μM ATPγS (for four to five days), or TNFα (8–12 h) and used for Western blotting.

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“… PBMCs were isolated from Leukopaks derived from healthy patients, then infected in vitro with HIV ADA 31 , 32 , 33 , 34 or pNL(AD 8 ) (10 ng/mL). 35 PBMCs (infected and uninfected) were either untreated or treated with 3 μM or 15 μM ATPγS (for four to five days), or TNFα (8–12 h) and used for Western blotting.…”

“… 7 BMVEC cultures were either exposed to ATPγS for at least seven passages. PBMCs infected in vitro with 10 ng/mL of either HIV ADA 31 , 32 , 33 , 34 or pNL(AD 8 ) (a CCR5-tropic molecular clone of NL4-3) 35 and after a single round of replication, about 18-24 hours after infection, ATPγS was added to the cultures for acute treatment of 4-5 days (5-6 days post-infection). For the co-culture assay, PBMCs were maintained in ATPγS for only 4 days before addition to the BMVEC culture.…”

Immunocapture of cell surface proteins embedded in HIV envelopes uncovers considerable virion genetic diversity associated with different source cell types

“…The depletion cocktail removed CD3, CD45RA, CD19, CD56, and CD235a cells. The monocyte-enriched cells were immediately placed into culture and inoculated with HIV-1 ADA-M [30] for two hours, then washed and replenished with media containing tumor necrosis factor (TNF)-α. Cell culture supernatant was collected after 2.5 days when viral RNA was detected.…”