SARS-CoV-2 Specific T-cell Assay

Splenocytes were incubated with SARS-CoV-2 S peptide pools (1 μg/mL, Miltenyi Biotec) for 6 h in the presence of GolgiPlug (BD Bioscience). Cells were harvested and stained with antibodies for CD3, CD4, or CD8, fixed in 2% paraformaldehyde, and permeabilized with 0.5% saponin before adding anti-IFN-γ (Thermo Fisher Scientific) (24 (link), 31 (link)). Samples were acquired by a C6 Flow Cytometer instrument. Dead cells were excluded based on forward and side light scatter. Data were analyzed with a CFlow Plus Flow Cytometer (BD Biosciences).

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Singh A., Adam A., A.d.i.t.i., Peng B.H., Yu X., Zou J., Kulkarni V.V., Kan P., Jiang W., Shi P.Y., Samir P., Cisneros I, & Wang T. (2024). A murine model of post-acute neurological sequelae following SARS-CoV-2 variant infection. Frontiers in Immunology, 15, 1384516.

Publication 2024

SARS-CoV-2 S peptide pools GolgiPlug anti-IFN-γ CFlow Plus Flow Cytometer

Corresponding Organization : The University of Texas Medical Branch at Galveston

Other organizations : Medical University of South Carolina

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Variable analysis

independent variables

SARS-CoV-2 S peptide pools (1 μg/mL, Miltenyi Biotec)

dependent variables

IFN-γ production

control variables

Incubation time (6 h)
Presence of GolgiPlug (BD Bioscience)
Antibodies for CD3, CD4, or CD8
2% paraformaldehyde fixation
0.5% saponin permeabilization
Anti-IFN-γ (Thermo Fisher Scientific) staining
Exclusion of dead cells based on forward and side light scatter

positive controls

Not specified

negative controls

Not specified

Annotations

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