The Influence of Cryopreservation on Cytokine Production by Human T Lymphocytes

Wang, Shengyuan; Hsu, Ming-Ling; Tzeng, Cheng‐Hwai; Hsu, Hui‐Chi; Ho, Chi-Kuan

doi:10.1006/cryo.1998.2094

Cited by 33 publications

(23 citation statements)

References 21 publications

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“…When cells are exposed to stressful environments, e.g., cooling, heating, radiation, or desiccation, cellular functions may be impaired. It has been observed that for some cells, preincubation in broth or liquid medium for some time after exposure to the stress may help repair the injuries (1,8,15,23); however, others doubt its effectiveness (7). In the present work, preincubation did not improve the recovery of cryopreserved BCG cells from cryoinjury.…”

Section: Discussioncontrasting

confidence: 60%

“…Common cryopreservation media, the effect of a cryoprotective agent (glycerol), and the cooling rate were studied. It has been suggested that transient incubation of cells in broth medium after exposure to stress environments, such as heating, freezing/thawing, or UV light, helps to repair the injury for some cell types (1,8,15,23). Therefore, the effect of transient incubation (preincubation) on cryopreserved BCG cells was also explored.…”

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confidence: 99%

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Cryopreservation of Mycobacterium tuberculosis Complex Cells

et al. 2012

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Successful long-term preservation of Mycobacterium tuberculosis cells is important for sample transport, research, biobanking, and the development of new drugs, vaccines, biomarkers, and diagnostics. In this report, Mycobacterium bovis bacillus CalmetteGuérin and M. tuberculosis H37Ra were used as models of M. tuberculosis complex strains to study cryopreservation of M. tuberculosis complex cells in diverse sample matrices at different cooling rates. Cells were cryopreserved in diverse sample matrices, namely, phosphate-buffered saline (PBS), Middlebrook 7H9 medium with or without added glycerol, and human sputum. The efficacy of cryopreservation was quantified by microbiological culture and microscopy with BacLight LIVE/DEAD staining. In all sample matrices examined, the microbiological culture results showed that the cooling rate was the most critical factor influencing cell viability. Slow cooling (a few degrees Celsius per minute) resulted in much higher M. tuberculosis complex recovery rates than rapid cooling (direct immersion in liquid nitrogen) (P < 0.05). Among the three defined cryopreservation media (PBS, 7H9, and 7H9 plus glycerol), there was no significant differential effect on viability (P ‫؍‬ 0.06 to 0.87). Preincubation of thawed M. tuberculosis complex cells in 7H9 broth for 20 h before culture on solid Middlebrook 7H10 plates did not help the recovery of the cells from cryoinjury (P ‫؍‬ 0.14 to 0.71). The BacLight LIVE/DEAD staining kit, based on Syto 9 and propidium iodide (PI), was also applied to assess cell envelope integrity after cryopreservation. Using the kit, similar percentages of "live" cells with intact envelopes were observed for samples cryopreserved under different conditions, which was inconsistent with the microbiological culture results. This implies that suboptimal cryopreservation might not cause severe damage to the cell wall and/or membrane but instead cause intracellular injury, which leads to the loss of cell viability.

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Section: Discussioncontrasting

confidence: 60%

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confidence: 99%

Cryopreservation of Mycobacterium tuberculosis Complex Cells

et al. 2012

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“…Cell aliquots below this viability were discarded. Although no experiments were performed to test the effect of freezing conditions on both cytokine/chemokine production and cell surface CCR expression, previous studies showed that cryopreservation of cells from blood and bronchoalveolar lavages does not affect the pattern of expression of these molecules and that such cells possessed an ability in cytokine production and CCR expression as good as that of the fresh cells (34,35).…”

Section: Freezing Procedures and Cell Storagementioning

confidence: 99%

Mycobacterium tuberculosisAntigens Specifically Modulate CCR2 and MCP-1/CCL2 on Lymphoid Cells from Human Pulmonary Hilar Lymph Nodes

Arias

Jaramillo

López

et al. 2007

The Journal of Immunology

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Macrophages and dendritic cells are involved in the immune response to Mycobacterium tuberculosis (Mtb). Such a response, although extensively studied using animal models and cells from human blood, has not been characterized in cells from pulmonary hilar lymph nodes (PHLN). We characterized populations of myeloid APC from PHLN and determined their expression of CCR2, CCR5, CCR7, CD40, CD54, CD80, and CD86 as well as the cytokine/chemokine microenvironment before and after purified protein derivative (PPD) and mannosilated lipoarabinomannan (ManLAM) stimulation. Results show that there are at least three APC populations in PHLN, defined as CD14highHLA-DRlow/−, CD14dimHLA-DRdim, and CD14−HLA-DRhigh/dendritic cells (DC), with the largest number represented by CD14dimHLA-DRdim cells (where dim indicates intermediate levels). CD14−HLA-DRhigh/DC expressed higher levels of costimulatory molecules and lower levels of CCR2 and CCR5, but all cell populations showed similar CCR7 levels. PPD and ManLAM specifically down-regulated CCR2 expression but not that of CCR5 and CCR7, and such down-regulation was observed on all APC populations. Mtb Ag did not affect the expression of costimulatory molecules. PPD but not ManLAM specifically induced MCP-1/CCL2 production, which was likely associated with the induction of IFN-γ because this cytokine was highly induced by PPD. We characterized, for the first time, different APC from human PHLN and show that Mtb Ag exert fine and specific regulation of molecules closely associated with the immune response to Mtb infection. Because knowledge of this response in secondary lymphoid tissues is still poorly understood in humans, such studies are necessary and important for a better understanding of lymphoid cell microenvironment and migrating capacities and their role in the immunopathogenesis of tuberculosis.

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“…The expression of certain functionally related molecules of particular relevance in HIV-1 infection, such as CD38, is also relatively stable following cryopreservation (23). Likewise, the potential for PBMC to be induced to proliferate, secrete cytokines, or exhibit antigenspecific or nonspecific lytic activity can be preserved in stored, frozen PBMC (7,11,12,18,27,30). However, these studies were always performed in individual laboratories, often using specimens from normal or non-HIV-infected donors.…”

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Preservation of Lymphocyte Immunophenotype and Proliferative Responses in Cryopreserved Peripheral Blood Mononuclear Cells from Human Immunodeficiency Virus Type 1-Infected Donors: Implications for Multicenter Clinical Trials

Reimann

Chernoff

Wilkening

et al. 2000

Clin Diagn Lab Immunol

121

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Human immunodeficiency virus type 1 (HIV-1) infection results in impaired immune function that can be measured by changes in immunophenotypically defined lymphocyte subsets and other in vitro functional assays. These in vitro assays may also serve as early indicators of efficacy when new therapeutic strategies for HIV-1 infection are being evaluated. However, the use of in vitro assays of immune function in multicenter clinical trials has been hindered by their need to be performed on fresh specimens. We assessed the feasibility of using cryopreserved peripheral blood mononuclear cells (PBMC) for lymphocyte immunophenotyping and for lymphocyte proliferation at nine laboratories. In HIV-1-infected patients with moderate CD4؉ lymphocyte loss, the procedures of density gradient isolation, cryopreservation, and thawing of PBMC resulted in significant loss of CD19 ؉ B cells but no measurable loss of total T cells or CD4 ؉ or CD8 ؉ T cells. No significant changes were seen in CD28 ؊ CD95 ؉ lymphocytes after cell isolation and cryopreservation. However, small decreases in HLA-DR ؉ CD38 ؉ lymphocytes and of CD45RA ؉ CD62L ؉ were observed within both the CD4 ؉ and CD8 ؉ subsets. Fewer than 10% of those specimens that showed positive PBMC proliferative responses to mitogens or microbial antigens lost their responsiveness after cryopreservation. These results support the feasibility of cryopreserving PBMC for immunophenotyping and functional testing in multicenter AIDS clinical trials. However, small changes in selected lymphocyte subsets that may occur after PBMC isolation and cryopreservation will need to be assessed and considered in the design of each clinical trial.Therapeutic approaches that reduce the rate of human immunodeficiency virus type 1 (HIV-1) replication result in fewer clinical events and prolong the survival of infected individuals (10,22). In addition to an increase in CD4 T cells, immune function as measured by in vitro assays also improves significantly in response to antiretroviral treatments. Following initiation of highly active antiretroviral therapy, functionally relevant lymphocyte subsets in blood begin to return to normal levels. In addition, in vitro proliferation and cytokine secretion by peripheral blood mononuclear cells (PBMC) also show improvement (4, 14, 16, 24). As a result, in vitro measurements of immune function are being explored as a substitute for clinical endpoints in therapeutic trials (21). Furthermore, quantitating immune function through the use of these assays will provide a better definition of the immune reconstitution that occurs during antiretroviral therapy and may advance our understanding of AIDS pathogenesis.The in vitro assays currently used to measure immune function are technically complex, prone to variability, and usually performed in real time on fresh specimens. The problems of performing these assays efficiently and reproducibly are compounded in multicenter clinical trials. In these clinical trials, specimens are collected over many months and at multiple l...

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The Influence of Cryopreservation on Cytokine Production by Human T Lymphocytes

Cited by 33 publications

References 21 publications

Cryopreservation of Mycobacterium tuberculosis Complex Cells

Cryopreservation of Mycobacterium tuberculosis Complex Cells

Mycobacterium tuberculosisAntigens Specifically Modulate CCR2 and MCP-1/CCL2 on Lymphoid Cells from Human Pulmonary Hilar Lymph Nodes

Preservation of Lymphocyte Immunophenotype and Proliferative Responses in Cryopreserved Peripheral Blood Mononuclear Cells from Human Immunodeficiency Virus Type 1-Infected Donors: Implications for Multicenter Clinical Trials

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