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Related Experiment Videos

Effects of cryopreservation on lymphocyte immunophenotype and function.

A Costantini1, S Mancini, S Giuliodoro

  • 1Service of Clinical Immunology, Department of Internal Medicine, Ancona University Medical School, Ancona, Italy.

Journal of Immunological Methods
|September 6, 2003
PubMed
Summary

Cryopreservation significantly alters peripheral blood mononuclear cells (PBMCs), impacting T cell subsets and chemokine receptor expression. For HIV-infected patients, studies on immune reconstitution or infectivity require fresh samples to avoid altered results.

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Area of Science:

  • Immunology
  • Cell Biology
  • Virology

Background:

  • Cryopreservation of peripheral blood mononuclear cells (PBMCs) is standard for minimizing variability in phenotypic and functional analyses.
  • Limited data exist on cryopreservation's effects on PBMCs, particularly in HIV-infected individuals, where lymphocyte assessment is critical.

Purpose of the Study:

  • To compare the effects of cryopreservation on PBMCs from healthy individuals and HIV-infected patients.
  • To evaluate cryopreservation-induced changes in T cell subsets, chemokine receptor expression, and antigen-specific responses.

Main Methods:

  • Comparison of fresh versus frozen/thawed (F/T) PBMCs from 19 healthy and 21 HIV-infected individuals.
  • Flow cytometry analysis of cell surface markers (CD62L, CD45RA, CD45RO, CCR5).

Related Experiment Videos

  • Assessment of T cell proliferation in response to HIV and recall antigens.

    • Main Results:

    • Cryopreservation decreased CD62L expression, reducing naive and central memory T cells.
    • Effector CD8+ T cells increased in healthy subjects but not in HIV-infected patients.
    • Significant decline in CC chemokine receptor 5 (CCR5) expression observed.
    • HIV-infected patients showed loss of proliferative responses to p24, CMV, and Influenza antigens post-cryopreservation.

    Conclusions:

    • Cryopreservation induces consistent changes in PBMCs from both healthy and HIV-infected individuals.
    • Immunological studies in HIV-infected patients, such as immune reconstitution or in vitro infectivity assays, should utilize fresh samples.