Generation of Cytotoxic T Cells Against Virus-Infected Human Brain Macrophages in a Murine Model of HIV-1 Encephalitis

Abstract

HIV-1 encephalitis (HIVE) and its associated dementia can occur in up to 20% of infected individuals, usually when productive viral replication in brain mononuclear phagocytes (macrophages and microglia) and depletion of CD4+ T lymphocytes are most significant. T cells control viral replication through much of HIV-1 disease, but how this occurs remains incompletely understood. With this in mind, we studied HIV-1-specific CTL responses in a nonobese diabetic (NOD)-SCID mouse model of HIVE. HIV-1-infected monocyte-derived macrophages (MDM) were injected into the basal ganglia after syngeneic immune reconstitution by HLA-A*0201-positive human PBL to generate a human PBL-NOD-SCID HIVE mouse. Engrafted T lymphocytes produced HIV-1gag- and HIV-1pol-specific CTL against virus-infected brain MDM within 7 days. This was demonstrated by tetramer staining of human PBL in mouse spleens and by IFN-γ ELISPOT. CD8, granzyme B, HLA-DR, and CD45R0 Ag-reactive T cells and CD79α-positive B cells migrated to and were in contact with human MDM in brain areas where infected macrophages were abundant. The numbers of productively infected MDM were markedly reduced (>85%) during 2 wk of observation. The human PBL-NOD-SCID HIVE mouse provides a new tool for studies of cellular immune responses against HIV-1-infected brain mononuclear phagocytes during natural disease and after vaccination.

Pharmacodynamics of long-acting folic acid-receptor targeted ritonavir-boosted atazanavir nanoformulations

“…For reconstitution 25 × 10 6 hu-PBL per mouse were injected intraperitoneally into 8–10 week old NOD/SCID/IL2Rγc −/− (NSG) mice obtained from a University of Nebraska Medical Center (UNMC) breeding colony under an approved UNMC Institutional Animal Care and Use protocol as previously described [22]. …”

Long-acting nanoformulated antiretroviral therapy elicits potent antiretroviral and neuroprotective responses in HIV-1-infected humanized mice

“…For immunohistochemical analysis, tissues were fixed with 4% paraformaldehyde and embedded in paraffin. Five-micron thick sections were immunostained for CD45LCA (Dako, Carpinteria, CA 1:200), HLA-DR (clone CR3/43, 1:100) and HIV-1 p24 (1:10)[13, 18]. Images were obtained with a Nuance EX camera fixed to a Nikon Eclipse E800 using Nuance software (Cambridge Research & Instrumentation, Woburn, MA).…”

Humanized mice for HIV and AIDS research

“…Direct injection of HIV-infected cells into the brains of mice results in neuropathology similar to humans with HIVE (HIV encephalitis), including encephalitis, astrogliosis, multinucleated giant cell formation, infiltration of mononuclear phagocytes, and decreased microtubule-associated protein-2 (MAP-2) expression [76,77]. In another model, mice previously reconstituted with human peripheral blood leukocytes (PBLs) were inoculated with HIV-infected autologous macrophages in the brain [78,79]. In these mice, human PBLs were found in the meninges, choroid plexus, and the ventricles which are regions patrolled by circulating immune cells [80].…”