Author: Banu, Nasirah; Chia, Adeline; Ho, Zi Zong; Garcia, Alfonso Tan; Paravasivam, Komathi; Grotenbreg, Gijsbert M.; Bertoletti, Antonio; Gehring, Adam J.
Title: Building and Optimizing a Virus-specific T Cell Receptor Library for Targeted Immunotherapy in Viral Infections Document date: 2014_2_25
ID: 44w6omdp_27
Snippet: Postive responses were restimulated to increase frequency and numbers of peptidespecific T cells. Peptide-specific T cell cultures (5 3 10 5 ) were co-cultured with irradiated (2500 rads) healthy donor PBMC (1.7 3 10 6 ) and HLA-matched EBV transformed B cell lines (3 3 10 5 ) loaded with peptide for 1 h at room temperature. Restimulated T cells were cultured in AIM-V media supplemented with 2% AB serum, 20 U/ml IL-2, 10 ng/ml IL-7 and 10 ng/ml I.....
Document: Postive responses were restimulated to increase frequency and numbers of peptidespecific T cells. Peptide-specific T cell cultures (5 3 10 5 ) were co-cultured with irradiated (2500 rads) healthy donor PBMC (1.7 3 10 6 ) and HLA-matched EBV transformed B cell lines (3 3 10 5 ) loaded with peptide for 1 h at room temperature. Restimulated T cells were cultured in AIM-V media supplemented with 2% AB serum, 20 U/ml IL-2, 10 ng/ml IL-7 and 10 ng/ml IL-15 (R&D Systems). Functionality was tested again 10 d after restimulation using the CD107a degranulation assay to maintain cell viability. The TCR variable beta chain staining panel IO Test Beta Mark TCR V kit (Beckman Coulter) was used to determine the immunodominant Vb on CD107a1 cells following peptide stimulation.
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