Enrichment of Ag-specific CD8+ T Cells

Enrichment of Ag-specific CD8⁺ T cells was performed using an established protocol^{22 (link)}. Combined spleen and lymph nodes were digested with collagenase D. Cells were then labeled with APC-conjugated H-2K^d-CSP^280–288 tetramer (SYVPSAEQI, NIH Tetramer Core Facility) for 30 min at room temperature, followed by magnetic enrichment over LS columns using anti-APC microbeads (Miltenyi biotec). Enriched samples and unbound fractions were stained with surface antibodies, and Polybead polystyrene microspheres (Polyscience) were used for calculating cell number.

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Rolot M., Dougall A.M., Chetty A., Javaux J., Chen T., Xiao X., Machiels B., Selkirk M.E., Maizels R.M., Hokke C., Denis O., Brombacher F., Vanderplasschen A., Gillet L., Horsnell W.G, & Dewals B.G. (2018). Helminth-induced IL-4 expands bystander memory CD8+ T cells for early control of viral infection. Nature Communications, 9, 4516.

Publication 2018

anti-APC microbeads Polybead polystyrene microspheres

Cd8 t cells Cells Collagenase Lymph nodes Microspheres Polystyrene Spleen Surface antibodies Tetramer

Corresponding Organization :

Other organizations : University of Liège, University of Cape Town, Imperial College London, Wellcome Centre for Molecular Parasitology, University of Glasgow, Leiden University Medical Center, Institut Scientifique de Santé Publique

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Variable analysis

independent variables

Enrichment of Ag-specific CD8+ T cells using an established protocol

dependent variables

Proportion of Ag-specific CD8+ T cells in the enriched samples

control variables

Spleen and lymph node cells
Collagenase D digestion
APC-conjugated H-2Kd-CSP280–288 tetramer staining
Magnetic enrichment using anti-APC microbeads
Surface antibody staining
Polybead polystyrene microspheres for cell counting

controls

Positive control: Not mentioned
Negative control: Not mentioned

Annotations

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