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Effects of cryopreservation on lymphocyte immunophenotype and function
Abstract
Cryopreservation of isolated peripheral blood mononuclear cells (PBMCs) for phenotypic and functional analyses is considered a standard procedure in order to minimize operator-dependent inter-assay variability and to optimize the use of available resources. However, only few and somewhat conflicting data are presently available on the effects of cryopreservation on PBMCs, especially in samples from HIV-infected patients in which assessment of lymphocyte phenotype and function is of the outmost importance. In this study, we compared fresh versus frozen/thawed (F/T) samples isolated from 19 healthy individuals and 21 HIV-infected patients, showing that cryopreservation induces: (i) a profound decrease of CD62L expression, with a consequent significant decline of the calculated proportions of "naïve" (CD45RA+CD62L+) and "central memory" (CD45RO+CD62L+) T cells; (ii) an increase of the calculated proportions of "effector" CD8+ T cells (CD45RA+CD62L- and CD45RO+CD62L-) in the healthy subjects, while no changes were observed in the HIV-infected group; (iii) a significant decline of CC chemokine receptor 5 (CCR5) expression; (iv) a loss of proliferative responses to some HIV antigens (i.e. p24) and recall antigens [cytomegalovirus (CMV) and Influenza] in HIV-infected patients. We thus conclude that cryopreservation induces a consistent set of changes in PBMCs from both healthy and HIV-infected individuals, and that certain immunological studies of HIV-infected patients (i.e. studies of immune reconstitution following antiretroviral therapy in HIV-infected patients or studies of HIV-infectivity in vitro using CCR5-tropic strains) should be performed on fresh samples.
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“although CD62L expression has been reported to be profoundly affected by cryopreservation [138] thereby limiting its use to freshly isolated cells.”
From Guidelines for the use of flow cytometry and cell sorting in immunological studies (third edition) (2021) · cites this paper as [138]
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Cites
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Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)
Cossarizza, A., Chang, H., Radbruch, A., Acs, A., Adam, D., Adam‐Klages, S., Agace, W. W., Aghaeepour, N., Akdiş, M., Allez, M., et al. (2019) · European Journal of Immunology · doi:10.1002/eji.201970107 · Cited by 1,007
Newell, E. W., Sigal, N., Bendall, S. C., Nolan, G. P., & Davis, M. M. (2012) · Immunity · doi:10.1016/j.immuni.2012.01.002 · Cited by 555
Guidelines for the use of flow cytometry and cell sorting in immunological studies *
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Yin, K., Peluso, M. J., Luo, X., Thomas, R., Shin, M., Neidleman, J. A., Andrew, A. K., Young, K. C., Ma, T., Hoh, R., et al. (2024) · Nature Immunology · doi:10.1038/s41590-023-01724-6 · Cited by 410
Metadata
- Authors
- Costantini, A., Mancini, S., Giuliodoro, S., Butini, L., Regnery, C. M., Silvestri, G., & Montroni, M.
- Affiliations
- Emory University (US)
- Azienda Ospedaliero-Universitaria delle Marche (IT)
- Corresponding author
- Montroni, M.
- Venue
- Journal of Immunological Methods
- Publisher
- Elsevier BV
- Published
- 2003-07
- Volume
- 278
- Issue
- 1-2
- Pages
- 145-155
- ISSN
- 0022-1759 (ISSN-L)
- OpenAlex
- W2060902798
- Cited by
- 185
- References
- 18
- Citations by year
- 2026: 3 · 2025: 10 · 2024: 7 · 2023: 12 · 2022: 9 · 2021: 15 · 2020: 13 · 2019: 6 · 2018: 15 · 2017: 7 · 2016: 8 · 2015: 7 · 2014: 13 · 2013: 9 · 2012: 7
- FWCI
- 2.00
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Immunological Methods (publishedVersion)
- PubMed (publishedVersion)
- Università Politecnica delle Marche (Università Politecnica delle Marche) (submittedVersion)
- Topics
- Life Sciences › Immunology and Microbiology › Virology › HIV Research and Treatment
- Life Sciences › Immunology and Microbiology › Immunology › T-cell and B-cell Immunology
- Health Sciences › Health Professions › Medical Laboratory Technology › Clinical Laboratory Practices and Quality Control
- Keywords
- cryopreservation, peripheral blood mononuclear cells, lymphocyte immunophenotype, HIV infection, L-selectin, CCR5, naive T cells, central memory T cells, CD8+ effector T cells, T cell proliferation, p24 antigen, cytomegalovirus, influenza, sample storage
- MeSH terms
- Humans, Immunity, Cellular, T-Lymphocytes, HIV Infections, Cryopreservation, Immunophenotyping, T-Lymphocyte Subsets, L-Selectin, Receptors, CCR5
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