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Cells 2017, 6(4), 47; doi:10.3390/cells6040047

A Positive Control for Detection of Functional CD4 T Cells in PBMC: The CPI Pool

1
Research&Development Department, Cellular Technology Limited, Shaker Heights, OH 44122, USA
2
Department of Cancer Immunology, Chair of Medical Biotechnology, Poznan University of Medical Sciences, 61-701 Poznan, Poland
3
Institute of Anatomy and Cell Biology, Friedrich-Alexander University Erlangen-Nürnberg, 1054 Erlangen, Germany
*
Author to whom correspondence should be addressed.
Received: 12 November 2017 / Revised: 1 December 2017 / Accepted: 3 December 2017 / Published: 7 December 2017
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Abstract

Testing of peripheral blood mononuclear cells (PBMC) for immune monitoring purposes requires verification of their functionality. This is of particular concern when the PBMC have been shipped or stored for prolonged periods of time. While the CEF (Cytomegalo-, Epstein-Barr and Flu-virus) peptide pool has become the gold standard for testing CD8 cell functionality, a positive control for CD4 cells is so far lacking. The latter ideally consists of proteins so as to control for the functionality of the antigen processing and presentation compartments, as well. Aiming to generate a positive control for CD4 cells, we first selected 12 protein antigens from infectious/environmental organisms that are ubiquitous: Varicella, Influenza, Parainfluenza, Mumps, Cytomegalovirus, Streptococcus, Mycoplasma, Lactobacillus, Neisseria, Candida, Rubella, and Measles. Of these antigens, three were found to elicited interferon (IFN)-γ-producing CD4 cells in the majority of human test subjects: inactivated cytomegalo-, parainfluenza-, and influenza virions (CPI). While individually none of these three antigens triggered a recall response in all donors, the pool of the three (the ‘CPI pool’), did. One hundred percent of 245 human donors tested were found to be CPI positive, including Caucasians, Asians, and African-Americans. Therefore, the CPI pool appears to be suitable to serve as universal positive control for verifying the functionality of CD4 and of antigen presenting cells. View Full-Text
Keywords: PBMC quality assessment; PBMC cryopreservation; CD4 cell function; ELISPOT; ImmunoSpot; immune monitoring PBMC quality assessment; PBMC cryopreservation; CD4 cell function; ELISPOT; ImmunoSpot; immune monitoring
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Schiller, A.; Zhang, T.; Li, R.; Duechting, A.; Sundararaman, S.; Przybyla, A.; Kuerten, S.; Lehmann, P.V. A Positive Control for Detection of Functional CD4 T Cells in PBMC: The CPI Pool. Cells 2017, 6, 47.

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