Isolation and Activation of CD8+ T Cells

Pure CD8⁺ T cells were isolated from healthy donor PBMC by using the EasySep naïve CD8⁺ T cell enrichment kit (#19158, StemCell Technologies, Vancouver, BC, Canada). Monocytes-grown dendritic cells were loaded with staphylococcal enterotoxin B (SEB, 1 ng/mL). The previously isolated CD8⁺ T cells were co-cultured with SEB-loaded dendritic cells for six days. On day 7, pure T cells were harvested and adjusted to 10⁶ cells/mL for running the chemotaxis assay [33 (link),47 (link)].

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Ibrahim O.M., Basse P.H., Jiang W., Guru K., Chatta G, & Kalinski P. (2021). NFκB-Activated COX2/PGE2/EP4 Axis Controls the Magnitude and Selectivity of BCG-Induced Inflammation in Human Bladder Cancer Tissues. Cancers, 13(6), 1323.

Publication 2021

EasySep naïve CD8+ T cell enrichment kit

Assay Cd8 t cells Cells Chemotaxis Dendritic cells Donor Monocytes Stemcell T cells

Corresponding Organization : Roswell Park Comprehensive Cancer Center

Other organizations : The Graduate Center, CUNY, University at Buffalo, State University of New York

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Variable analysis

independent variables

SEB (1 ng/mL) loading of dendritic cells

dependent variables

Chemotaxis of CD8+ T cells

control variables

Isolation of pure CD8+ T cells from healthy donor PBMC
Co-culture of CD8+ T cells with SEB-loaded dendritic cells for six days
Harvesting of pure T cells on day 7 and adjusting to 10^6 cells/mL for chemotaxis assay

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