Visualizing SINV Antibody Clearance in Infected Cells

For analysis of immune-mediated virus clearance mouse monoclonal IgG3 antibody against SINV E2 (SV127; 209^{32 (link)}) was used. For antibody imaging, SV127 was conjugated with Dylight 594 (Thermo Scientific, ~5 dye molecules/IgG molecule) as previously described^{20 (link)}. Infected cells were treated with 1–5 μg/ml of SV127 or 500 U/ml of rat IFNγ (PBL). Antibody to TrkA (Clone 315104, R&D Systems) at 1 μg/ml was used as a control. Infected cells were identified with a rabbit polyclonal antibody to SINV nsP3^{33 (link)} and CF633-conjugated anti rabbit IgG(H + L) (Biotium).

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Nilaratanakul V., Hauer D.A, & Griffin D.E. (2020). Visualization of cell-type dependent effects of anti-E2 antibody and interferon-gamma treatments on localization and expression of Broccoli aptamer-tagged alphavirus RNAs. Scientific Reports, 10, 5259.

Publication 2020

Dylight 594

Anti igg Antibody Cells Clone Ifnγ Igg3 Monoclonal antibody Mouse Rabbit Virus

Corresponding Organization :

Other organizations : Johns Hopkins University

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Variable analysis

independent variables

Antibody concentration (1-5 μg/ml of SV127)

dependent variables

Immune-mediated virus clearance

control variables

Infected cells treated with 500 U/ml of rat IFNγ
Infected cells treated with 1 μg/ml of antibody to TrkA (Clone 315104, R&D Systems)

positive controls

Infected cells treated with 500 U/ml of rat IFNγ

negative controls

Infected cells treated with 1 μg/ml of antibody to TrkA (Clone 315104, R&D Systems)

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