HIV-infected Macrophage Isolation and Characterization

MDM were infected with HIV NL4.3 BAL-IRES-HSA for 6 days at 37°C with 5% CO₂. HIV-1 infection was confirmed by quantitative PCR (qPCR) (66 (link)), p24 ELISA, and surface expression of virus-encoded murine heat-stable antigen (HSA) by flow cytometry (24 (link)). HSA-expressing MDM were isolated by positive selection using Miltenyi LS columns in combination with the MidiMACS magnet (Miltenyi Biotech). The HSA sorting protocol was optimized by the laboratory of Michel Tremblay, as described previously (24 (link), 25 (link)). Purity of the HSA⁺ and HSA⁻ fractions was assessed by flow cytometry.

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Sandstrom T.S., Ranganath N., Burke Schinkel S.C., Salahuddin S., Meziane O., Côté S.C., Costiniuk C.T., Jenabian M.A, & Angel J.B. (2021). HIV-Infected Macrophages Are Infected and Killed by the Interferon-Sensitive Rhabdovirus MG1. Journal of Virology, 95(9), e01953-20.

Publication 2021

Miltenyi LS columns MidiMACS magnet

Antigen Antigen surface Elisa Flow cytometry Hiv infection Ires Murine Virus

Corresponding Organization : Ottawa Hospital

Other organizations : McGill University Health Centre, Université du Québec à Montréal, McGill University, Université de Montréal

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Variable analysis

independent variables

HIV NL4.3 BAL-IRES-HSA infection

dependent variables

Quantitative PCR (qPCR) for HIV-1 infection
P24 ELISA for HIV-1 infection
Surface expression of virus-encoded murine heat-stable antigen (HSA) by flow cytometry
Purity of the HSA+ and HSA- fractions assessed by flow cytometry

control variables

Incubation of MDM at 37°C with 5% CO2 for 6 days

positive controls

Not explicitly mentioned

negative controls

Not explicitly mentioned

Annotations

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