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Cryopreservation modulates the detection of regulatory T cell markers

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dc.contributor.author Sattui, S.
dc.contributor.author De La Flor, C.
dc.contributor.author Sanchez, C.
dc.contributor.author Lewis, D.
dc.contributor.author Lopez, G.
dc.contributor.author Rizo-Patrõn, E.
dc.contributor.author White Jr., A.C.
dc.contributor.author Montes Delgado, Martin
dc.date.accessioned 2022-01-18T19:26:44Z
dc.date.available 2022-01-18T19:26:44Z
dc.date.issued 2012
dc.identifier.uri https://hdl.handle.net/20.500.12866/10815
dc.description.abstract Background: Regulatory T cells (Tregs) modulate the host response in infectious diseases and are key mediators of peripheral tolerance. Cryopreservation of peripheral blood mononuclear cells (PBMCs) is commonly used in immunological field studies where access to complex laboratory tests is not feasible. Our objective is to assess the effects of cryopreservation on the flow cytometric detection of surface and intracellular markers of Tregs. Methods: Heparinized venous blood was obtained from 36 healthy individuals and 15 HIV-1 infected subjects. PBMCs were isolated and stained for surface and intracellular markers of Tregs. PBMCs from each subject were cryopreserved in liquid nitrogen with DMSO; these cells were thawed and stained at a later date. All samples were analyzed by flow cytometry. The proportion of Tregs was compared using Wilcoxon signed-rank test. Results: Cryopreservation decreased the proportion of Tregs identified by surface and intracellular markers in healthy individuals and in HIV-1 patients. The proportion of CD4+CD25+FoxP3+ was decreased from 3.13 to 2.16% (P < 0.001) for non-HIV subjects and from 2.68 to 0.94% (P < 0.001) for HIV subjects, compared to fresh samples. Significant reduction was also observed for CD4+CD25+CD127lo-neg. However, the effect varied considerably between samples. The effect was similar among HIV and non-HIV patients (P = 0.38). Conclusions: Cryopreservation modulates the detection of surface and intracellular markers of Tregs. These results confirm that research on Tregs, including studies of HIV-1 infected patients, should be carried out prospectively on fresh samples in order to obtain unbiased conclusions. Results using cryopreserved cells should be regarded as only preliminary. en_US
dc.language.iso eng
dc.publisher Wiley
dc.relation.ispartofseries Cytometry Part B - Clinical Cytometry
dc.rights info:eu-repo/semantics/restrictedAccess
dc.rights.uri https://creativecommons.org/licenses/by-nc-nd/4.0/deed.es
dc.subject Humans en_US
dc.subject Controlled Study en_US
dc.subject HIV Infections en_US
dc.subject Clinical |Flow Cytometry en_US
dc.subject Peripheral Blood Mononuclear Cell en_US
dc.subject Biological Markers en_US
dc.subject Human Immunodeficiency Virus Infected Patient en_US
dc.subject Regulatory T Lymphocyte en_US
dc.subject Transcription Factor FOXP3 en_US
dc.subject Artifacts en_US
dc.subject CD4+ CD25+ T Lymphocyte en_US
dc.subject Cold Temperature en_US
dc.subject Cryopreservation en_US
dc.subject Flow Cytometry en_US
dc.subject Forkhead Transcription Factors en_US
dc.subject Foxp3 en_US
dc.subject Human Immunodeficiency Virus 1 Infection en_US
dc.subject Immunophenotyping en_US
dc.subject Interleukin-2 Receptor Alpha Subunit en_US
dc.subject Interleukin-7 Receptor Alpha Subunit en_US
dc.subject Leukocytes Mononuclear en_US
dc.subject Lymphocyte Surface Marker en_US
dc.subject T-Lymphocytes Regulatory en_US
dc.title Cryopreservation modulates the detection of regulatory T cell markers en_US
dc.type info:eu-repo/semantics/article
dc.identifier.doi https://doi.org/10.1002/cyto.b.20621
dc.subject.ocde https://purl.org/pe-repo/ocde/ford#3.01.03
dc.relation.issn 1552-4957


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