Effects of cryopreservation on lymphocyte immunophenotype and function

Costantini, Andrea; Mancini, Stefania; Giuliodoro, Simona; Butini, Luca; Regnery, C.M.; Silvestri, Guido; Montroni, M

doi:10.1016/s0022-1759(03)00202-3

Cited by 164 publications

(155 citation statements)

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“…The disadvantage of combining two different interventions in one treatment arm is that we are not able to distinguish separate effects for each intervention (partially hydrolysed formula and prebiotic supplementation) – it is therefore possible, albeit unlikely, that the interventions interacted to lead to no clinical effect, whereas separate use of one of the interventions might have resulted in a different outcome. Our analyses of PBMC phenotype and function were limited by relatively small numbers, and findings must be interpreted cautiously as they were not corrected for multiple comparisons, and the study set‐up necessitated the use of cryopreserved cells which can influence PBMC phenotype and function 29. Our study took place in a variety of settings in different countries, so the findings are likely to be generalizable to a range of settings.…”

Section: Discussionmentioning

confidence: 99%

Prebiotic‐supplemented partially hydrolysed cow's milk formula for the prevention of eczema in high‐risk infants: a randomized controlled trial

Boyle

Tang

Chiang

et al. 2016

Allergy

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BackgroundPrevention guidelines for infants at high risk of allergic disease recommend hydrolysed formula if formula is introduced before 6 months, but evidence is mixed. Adding specific oligosaccharides may improve outcomes.ObjectiveTo evaluate whether partially hydrolysed whey formula containing oligosaccharides (0.8 g/100 ml) (pHF‐OS) can prevent eczema in high‐risk infants [ISRCTN65195597].MethodsWe conducted a parallel‐group, multicentre, randomized double‐blind controlled trial of pHF‐OS vs standard cow's milk formula. Infants with a family history of allergic disease were randomized (stratified by centre/maternal allergy) to active (n = 432) or control (n = 431) formula until 6 months of age if formula was introduced before 18 weeks. Primary outcome was cumulative incidence of eczema by 12 months in infants randomized at 0–4 weeks (375 pHF‐OS, 383 control). Secondary outcomes were cumulative incidence of eczema by 12 or 18 months in all infants randomized, immune markers at 6 months and adverse events.ResultsEczema occurred by 12 months in 84/293 (28.7%) infants allocated to pHF‐OS at 0‐4 weeks of age, vs 93/324 (28.7%) control (OR 0.98 95% CI 0.68, 1.40; P = 0.90), and 107/347 (30.8%) pHF‐OS vs 112/370 (30.3%) control in all infants randomized (OR 0.99 95% CI 0.71, 1.37; P = 0.94). pHF‐OS did not change most immune markers including total/specific IgE; however, pHF‐OS reduced cow's milk‐specific IgG1 (P < 0.0001) and increased regulatory T‐cell and plasmacytoid dendritic cell percentages. There was no group difference in adverse events.ConclusionpHF‐OS does not prevent eczema in the first year in high‐risk infants. The immunological changes found require confirmation in a separate cohort.

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Section: Discussionmentioning

confidence: 99%

Prebiotic‐supplemented partially hydrolysed cow's milk formula for the prevention of eczema in high‐risk infants: a randomized controlled trial

Boyle

Tang

Chiang

et al. 2016

Allergy

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“…Cryopreservation is known to alter the expression of surface markers used to define T cell phenotype, such as CD62L (Tollerud et al, 1991;Romeu et al, 1992;Rosillo et al, 1995;Koenigsmann et al, 1998;Hattori et al, 2001;Cavers et al, 2002;Costantini et al, 2003). Our study avoided such markers and used others (CD28 and CCR7) that are not lost on previously frozen PBMC (Hamann et al, 1997;Hislop et al, 2001;Palmer et al, 2004;Yue et al, 2004).…”

Section: Discussionmentioning

confidence: 99%

“…Our study avoided such markers and used others (CD28 and CCR7) that are not lost on previously frozen PBMC (Hamann et al, 1997;Hislop et al, 2001;Palmer et al, 2004;Yue et al, 2004). Costantini et al (2003) reported an increase in CD45RA expression on cryopreserved PBMC. This may account for the increase in effector memory T cells measured, defined as CD45RA +/− CD28 − .…”

Section: Discussionmentioning

confidence: 99%

“…DMSO at such concentrations is toxic to cells. The risk of damage to cells from the freezing and thawing process is high and may result in alterations to the phenotype and function of cells (Tollerud et al, 1991;Romeu et al, 1992;Rosillo et al, 1995;Koenigsmann et al, 1998;Hattori et al, 2001;Cavers et al, 2002;Costantini et al, 2003). The lethality of intermediate temperatures during cooling and thawing are particularly problematic (Gao and Critser, 2000), and research groups have optimized freezing and storage conditions to offer preservation of cell viability (Kreher et al, 2003;Disis et al, 2006;Smith et al, 2007).…”

Section: Introductionmentioning

confidence: 99%

“…Other investigators have also measured an improvement of responses in frozen samples versus freshly isolated PBMC (Weinberg et al, 1998;Maecker et al, 2005). However, there are also reports of reduced function assessed by lymphoproliferative assays to select recall antigens such as HIV p24 and CMV, and to mitogens in frozen PBMC compared to freshly isolated PBMC (Costantini et al, 2003;Miniscalco et al, 2003). The reason for these inconsistent observations has not been well defined.…”

Section: Introductionmentioning

confidence: 99%

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Loss of T cell responses following long-term cryopreservation

Owen

Sinclair

Emu

et al. 2007

Journal of Immunological Methods

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Although cryopreservation of peripheral blood mononuclear cells (PBMC) is a commonly used technique, the degree to which it affects subsequent functional studies has not been well defined.Here we demonstrate that long-term cryopreservation has detrimental effects on T cell IFN-γ responses in human immunodeficiency virus (HIV) infected individuals. Long-term cryopreservation caused marked decreases in CD4 + T cell responses to whole proteins (HIV p55 and cytomegalovirus (CMV) lysate) and HIV peptides, and more limited decreases in CD8 + T cell responses to whole proteins. These losses were more apparent in cells stored for greater than one year compared to less than six months. CD8 + T cell responses to peptides and peptide pools were well preserved. Loss of both CD4 + and CD8 + T cell responses to CMV peptide pools were minimal in HIV-negative individuals. Addition of exogenous antigen presenting cells (APC) did not restore CD4 + T cell responses to peptide stimulation and partially restored T cell IFN-γ responses to p55 protein.Overnight resting of thawed cells did not restore T cell IFN-γ responses to peptide or whole protein stimulation. A selective loss of phenotypically defined effector cells did not explain the decrement of responses, although cryopreservation did increase CD4 + T cell apoptosis, possibly contributing to the loss of responses. These data suggest that the impact of cryopreservation should be carefully considered in future vaccine and pathogenesis studies. In HIV-infected individuals short-term cryopreservation may be acceptable for measuring CD4 + and CD8 + T cell responses. Long-term cryopreservation, however, may lead to the loss of CD4 + T cell responses and mild skewing of T cell phenotypic marker expression.

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Long‐term peripheral immune cell profiling reveals further targets of oral cladribine in MS

Moser

Schwenker

Seiberl

et al. 2020

Ann Clin Transl Neurol

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Objectives: To expand the knowledge about the immunological consequences of cladribine (CLAD), a pulsed immune reconstitution therapy approved for active multiple sclerosis (MS), beyond the known short-term effects on peripheral immune cell subsets. Methods: In this study, we characterized depletion and restitution kinetics as well as cytokine profiles of peripheral immune cell subsets in 18 patients with MS following treatment with oral CLAD. The methods involved blood collection prior to CLAD and every three months over a period of 24 months, and extensive characterization of various immune cells subsets by multiparametric flow cytometry. Results: We found a selectivity of CLAD towards central memory T cells and memory B cells and detected a hyper-repopulation of maturing B cells. Counts of classical (À65%) and various nonclassical TH17 cells (À84% to À87%) were markedly reduced 24 months after treatment start, and were comparable with depletion rates of classswitched memory B-cell phenotypes (À87% to À95%). The nadir of TH cells was more pronounced in the second treatment year. We observed a proportional surge of CD20 T-cell subsets and an expansion of regulatory T, B and NK cells. Natural killer T cells (NKT) were only depleted in year two and did not recover. Interpretation: Peripheral immune cell profiling revealed more differentiated insights into the immunological effects of CLAD. While some immune cell subsets expanded, we also observed additive depleting effects after the second treatment course. Further studies are required to elucidate whether these changes are paramount for the consistent and prolonged disease-modifying effect of CLAD.

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Effects of cryopreservation on lymphocyte immunophenotype and function

Cited by 164 publications

References 18 publications

Prebiotic‐supplemented partially hydrolysed cow's milk formula for the prevention of eczema in high‐risk infants: a randomized controlled trial

Prebiotic‐supplemented partially hydrolysed cow's milk formula for the prevention of eczema in high‐risk infants: a randomized controlled trial

Loss of T cell responses following long-term cryopreservation

Long‐term peripheral immune cell profiling reveals further targets of oral cladribine in MS

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