期刊
JOURNAL OF IMMUNOLOGICAL METHODS
卷 385, 期 1-2, 页码 90-95出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2012.07.017
关键词
Artificial antigen-presenting cells; Protein coating protocol; Quality control by flow cytometry; T-cell activation
资金
- Ludwig Institute for Cancer Research
- Swiss Cancer League [02836-08-2011]
- Swiss National Science Foundation [310030_135553]
- Swiss National Center of Competence in Research (NCCR) Molecular Oncology
- Swiss National Science Foundation (SNF) [310030_135553] Funding Source: Swiss National Science Foundation (SNF)
Artificial antigen-presenting cells (aAPC) are widely used for both clinical and basic research applications, as cell-based or bead-based scaffolds, combining immune synapse components of interest. Adequate and controlled preparation of aAPCs is crucial for subsequent immunoassays. We reveal that certain proteins such as activatory anti-CD3 antibody can be out-competed by other proteins (e.g. inhibitory receptor ligands such as PDL1:Fc) during the coating of aAPC beads, under the usually performed coating procedures. This may be misleading, as we found that decreased CD8 T cell activity was not due to inhibitory receptor triggering but rather because of unexpectedly low anti-CD3 antibody density on the beads upon co-incubation with inhibitory receptor ligands. We propose an optimized protocol, and emphasize the need to quality-control the coating of proteins on aAPC beads prior to their use in immunoassays. (C) 2012 Elsevier B.V. All rights reserved.
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