Cryopreservation of monocytes or differentiated immature DCs leads to an altered cytokine response to TLR agonists and microbial stimulation

Meijerink, Marjolein; Ulluwishewa, Dulantha; Anderson, Rachel C.; Wells, Jerry M.

doi:10.1016/j.jim.2011.08.010

Cited by 23 publications

(16 citation statements)

References 16 publications

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“…However, the effects of cryopreservation on CD14 + cells remain unclear. The freeze/thaw cycle procedure may modify the differentiation of DCs and the subsequent response of these cells to inflammatory stimuli or pathogens [12].…”

Section: Discussionmentioning

confidence: 99%

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Immature Dendritic Cells Generated from Cryopreserved Human Monocytes Show Impaired Ability to Respond to LPS and to Induce Allogeneic Lymphocyte Proliferation

Silveira

Wowk

Machado³

et al. 2013

PLoS ONE

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Dendritic cells play a key role in the immune system, in the sensing of foreign antigens and triggering of an adaptive immune response. Cryopreservation of human monocytes was investigated to understand its effect on differentiation into immature monocyte-derived dendritic cells (imdDCs), the response to inflammatory stimuli and the ability to induce allogeneic lymphocyte proliferation. Cryopreserved (crp)-monocytes were able to differentiate into imdDCs, albeit to a lesser extent than freshly (frh)-obtained monocytes. Furthermore, crp-imdDCs had lower rates of maturation and cytokine/chemokine secretion in response to LPS than frh-imdDCs. Lower expression of Toll-like receptor 4 (at 24 and 48 h) and higher susceptibility to apoptosis in crp-imdDCs than in fresh cells would account for the impaired maturation and cytokine/chemokine secretion observed. A mixed leukocyte reaction showed that lymphocyte proliferation was lower with crp-imdDCs than with frh-imdDCs. These findings suggested that the source of monocytes used to generate human imdDCs could influence the accuracy of results observed in studies of the immune response to pathogens, lymphocyte activation, vaccination and antigen sensing. It is not always possible to work with freshly isolated monocytes but the possible effects of freezing/thawing on the biology and responsiveness of imdDCs should be taken into account.

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Section: Discussionmentioning

confidence: 99%

“…CD14 + cells were cryopreserved (crp-CD14 + ) in inactivated fetal bovine serum (iFBS; Gibco-BRL, Grand Island, NY) containing 10% DMSO (Sigma-Aldrich, St. Louis, MO) as previously described [12], [16]. CD14 + cells were thawed by incubation for 1 to 2 min at 37°C in a water bath, and washed with iFBS by centrifugation at 300× g for 10 min.…”

Section: Methodsmentioning

confidence: 99%

Immature Dendritic Cells Generated from Cryopreserved Human Monocytes Show Impaired Ability to Respond to LPS and to Induce Allogeneic Lymphocyte Proliferation

Silveira

Wowk

Machado³

et al. 2013

PLoS ONE

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“…Hence, the properties of cells that have experienced freezing-thawing cycle need to be fully addressed. Several studies in the early 2000s and recent years have described the effect of cryopreservation on the biology and function of DCs in vitro or in vivo , however, some conclusions are controversial10111213 and most importantly, up to now, there is still a lack of confirming evidence on whether cryopreservation would have an effect on DCs’ homing capacity.…”

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confidence: 99%

Mature dendritic cell derived from cryopreserved immature dendritic cell shows impaired homing ability and reduced anti-viral therapeutic effects

Zhou

Zhao

Wang

et al. 2016

Sci Rep

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Cryopreservation is critical in reducing redundant operations and also in quality control in dendritic cell (DC) therapy. Full maturation and efficient homing of DCs to T cell-region constitute a crucial aspect of DC immunotherapy; however, the in vivo migration and distribution pattern, as well as the anti-viral effect of DCs that matured from cryopreserved immature DCs (cryoim-mDCs) remain to be revealed. In the present study, we compared cryoim-mDCs with DCs matured from fresh immature DCs (fmDCs) in the aspects of phenotypes, in vivo homing capacities as well as the anti-viral therapeutic effects to further clarify the effect of cryopreservation on DC-based cytotherapy. The results showed that cryopreservation impaired the homing ability of DCs which was associated with the reduced expression of CCR7 and disturbed cytoskeleton arrangement. Moreover, the antigen-specific CD8+ T cell response induced by cryoim-mDCs was much weaker than that induced by fmDCs in both the spleen and liver draining lymph nodes, which provided reduced protection from viral invasions. In conclusion, cryopreservation is a good method to keep the viability of immature DCs, however, the in vivo homing capacity and anti-viral therapeutic effect of DCs matured from frozen immature DCs were hindered to some extent.

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“…Generally, mature DCs are cryopreserved for this purpose . Some investigators attempted to freeze precursors of DCs, but these precursors after revival and maturation gave impaired DC phenotype as compared to their fresh counterparts …”

Section: Discussionmentioning

confidence: 99%

“…Large‐scale generation of DCs and their efficient cryopreservation, to be used as a ready‐to‐use frozen aliquot, would drastically increase the feasibility of DC vaccine. Though several studies have addressed the effect of cryopreservation on DC precursors and immature and tumor antigen–pulsed mature DCs, these studies have yielded contradictory results and still have lacunae . Dendritic cells, which play the role of professional antigen‐presenting cells, interact with T cells through tight cell membrane contact, to form an immunological synapse.…”

mentioning

confidence: 99%

Freezing of dendritic cells with trehalose as an additive in the conventional freezing medium results in improved recovery after cryopreservation

et al. 2018

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BACKGROUND: Dendritic cell (DC) vaccination involves administration of multiple doses. Cryopreservation of tumor antigen-pulsed DCs can provide a ready to use vaccine source and eliminate the need of frequent withdrawal of the patient's blood for vaccine preparation. The aim of this study was to assess the effect of addition of trehalose in the freezing medium on the recovery of DCs after cryopreservation.STUDY DESIGN AND METHODS: DCs were generated from mononuclear cells from apheresis samples of healthy donors. For long-term storage of 6 months, cells were frozen with a rate-controlled programmable freezer and stored in liquid nitrogen. For short-term storage of 1 month, cells were frozen and stored at −80 C. DCs frozen with Iscove's Modified Dulbecco's Medium + 10% dimethyl sulfoxide + 20% fetal bovine serum served as the control group, while the test group was additionally supplemented with 50 μg/mL of trehalose. After revival of control and test DCs, they were assessed for viability, morphology, phenotype, and functions. RESULTS:The addition of trehalose to the conventional freezing medium helped to preserve the viability and functionality of DCs better than dimethyl sulfoxide alone in both long-and short-term cryopreservation. Trehalose also protected the mitochondrial membrane potential and cytoskeleton integrity of DCs, which are necessary for their functionality. Mediators of the intrinsic apoptotic pathway like Caspase-9 and Bim-1 were found to be low in the test. CONCLUSION: Supplementation of conventional freezing medium with trehalose results in better quality of DCs revived after cryopreservation. This finding could help improve DC vaccine preparation for cancer immunotherapy. ABBREVIATIONS: CFSE = carboxyfluorescein succinimidyl ester; CPAs = cryoprotective agents; CTL = cytotoxic T-lympocyte; DC = dendritic cell; DMSO = dimethyl sulfoxide; FBS = fetal bovine serum; HLA = human leukocyte antigen; IMDM = Iscove's Modified Dulbecco's Medium; NCCS = National Centre for Cell Science; PBS = phosphate-buffered saline; PCR = polymerase chain reaction; TMRE = tetramethylrhodamine, ethyl ester. From the

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Cryopreservation of monocytes or differentiated immature DCs leads to an altered cytokine response to TLR agonists and microbial stimulation

Cited by 23 publications

References 16 publications

Immature Dendritic Cells Generated from Cryopreserved Human Monocytes Show Impaired Ability to Respond to LPS and to Induce Allogeneic Lymphocyte Proliferation

Immature Dendritic Cells Generated from Cryopreserved Human Monocytes Show Impaired Ability to Respond to LPS and to Induce Allogeneic Lymphocyte Proliferation

Mature dendritic cell derived from cryopreserved immature dendritic cell shows impaired homing ability and reduced anti-viral therapeutic effects

Freezing of dendritic cells with trehalose as an additive in the conventional freezing medium results in improved recovery after cryopreservation

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