Peripheral Blood Mononuclear Cells

Top PDF Peripheral Blood Mononuclear Cells:

Bovine Herpesvirus 1-Induced Apoptosis: Phenotypic Characterization of Susceptible Peripheral Blood Mononuclear Cells

Bovine Herpesvirus 1-Induced Apoptosis: Phenotypic Characterization of Susceptible Peripheral Blood Mononuclear Cells

In a variety of acute viral infections, there is increasing evidence that immunodepression is directly associated with the induction of apoptosis in immune cells [16, 19]. Apoptotic cell death is a genetically encoded suicide program which allows for the elimination of cells that have been produced in excess, developed improperly, or sustained genetic damage [23]. We have previously shown that BHV-1, even when inactivated, is able to induce apoptosis in mitogen- stimulated peripheral blood mononuclear cells (PBMCs) [11]. Apoptotic cell death is characterized morphologically by cell shrinkage, apoptotic body formation, condensation of the chromatin [6, 15], and biochemically by fragmentation of DNA into oligonucleosomal DNA fragments [1, 25]. Furthermore, the early stages of apoptosis are associated with the translocation of phosphatidylserine (PS) from the inner side of the plasma membrane to the outer layer [7]. This exposure of PS on the external surface of the cell membrane can be detected by the use of annexin V which is a Ca 2+ -dependent phospholipid-binding protein [18, 24].
En savoir plus

13 En savoir plus

Intravenous immunoglobulin mediates anti-inflammatory effects in peripheral blood mononuclear cells by inducing autophagy

Intravenous immunoglobulin mediates anti-inflammatory effects in peripheral blood mononuclear cells by inducing autophagy

Results IVIG induces autophagy in peripheral blood mononuclear cells of healthy donors In order to investigate whether IVIG induces autophagy, we first resorted to peripheral blood mononuclear cells (PBMCs) of healthy donors. PBMCs were cultured with different doses of IVIG (10 and 25 mg/ml) for 24 h and analyzed by western blot for the level of LC3-II, one of the well-characterized markers of autophagy 25 . Untreated PBMCs showed minimal level of LC3-II and its expression was not altered by equimolar concentrations of human serum albumin (HSA), an irrelevant protein control for IVIG. However, treatment of PBMCs with IVIG led to signi ficantly enhanced level of LC3-II (Fig. 1 a). Further, the effect of IVIG on autophagy was dose-dependent. High level of LC3-II was observed at 25 mg/ml, a con- centration of IVIG reaches in the circulation of auto- immune patients immediately following therapy. As LC3- II was also enhanced in cells treated with 10 mg
En savoir plus

15 En savoir plus

IRES complexity before IFN-alpha treatment and evolution of the viral load at the early stage of treatment in peripheral blood mononuclear cells from chronic hepatitis C patients.

IRES complexity before IFN-alpha treatment and evolution of the viral load at the early stage of treatment in peripheral blood mononuclear cells from chronic hepatitis C patients.

Shimazaki T, Honda M, Kaneko S, Kobayashi K. 2002. Inhibition of internal ribosomal entry site-directed translation of HCV by recombinant IFN-alpha correlates with a reduced La protein. Hepatology 35: 199-208. Shimizu YK, Igarashi H, Kanematu T, Fujiwara K, Wong DC, Purcell RH, Yoshikura H. 1997. Sequence analysis of the hepatitis C virus genome recovered from serum, liver, and peripheral blood mononuclear cells of infected chimpanzees. J Virol. 71: 5769-73. Shimizu YK, Purcell RH, Yoshikura H. 1993. Correlation between the infectivity of hepatitis
En savoir plus

27 En savoir plus

Clinical and kinomic analysis identifies peripheral blood mononuclear cells as a potential pharmacodynamic biomarker in metastatic renal cell carcinoma patients treated with sunitinib

Clinical and kinomic analysis identifies peripheral blood mononuclear cells as a potential pharmacodynamic biomarker in metastatic renal cell carcinoma patients treated with sunitinib

The use of kinase array technology to evaluate global kinase activities implied in cell signaling (« kinome ») in tumors from patients and to seek biomarkers for kinase inhibitors [3-7] is currently spreading. Recently, Anderson et al. have showed that the kinase profiling of clear cell RCC tumors could provide a functional classification strategy before starting a kinase inhibitor therapy [8]. Although these results are very promising, this strategy to process biopsy tumor before treatment initiation may fail to reflect current tumor dynamics and drug sensitivity, which may change during therapy. Additionally, processing repeated tumor biopsy cannot routinely be performed over the treatment course because of the invasive characteristic of the technique. Circulating cells could be an ideal biological matrix for biomarker discovery. In this context, we hypothesized that peripheral blood mononuclear cells (PBMC) could be a perfect surrogate tissue for identifying and assaying pharmacodynamic biomarkers of kinase inhibitors, as this tissue is readily accessible for repeat sampling throughout therapy. To the best of our knowledge, no data is currently available about the use of PBMC as surrogate tissue for the evaluation of global kinase activity in cancer patients.
En savoir plus

15 En savoir plus

HIV-1 DNA in peripheral blood mononuclear cells is strongly associated with HIV-1 disease progression in recently infected West African adults.

HIV-1 DNA in peripheral blood mononuclear cells is strongly associated with HIV-1 disease progression in recently infected West African adults.

HIV-1 DNA in peripheral blood mononuclear cells is strongly associated with HIV-1 disease progression in recently infected West African adults. Albert Minga, Xavier Anglaret, Thomas-d’Aquin Toni, Marie-Laure Chaix, Lambert Dohoun, Yao Abo, Ali Coulibaly, Julien Duvignac, Delphine

18 En savoir plus

Generation of the induced pluripotent stem cell line UHOMi002-A from peripheral blood mononuclear cells of a healthy male donor

Generation of the induced pluripotent stem cell line UHOMi002-A from peripheral blood mononuclear cells of a healthy male donor

Generation of the induced pluripotent stem cell line UHOMi002-A from peripheral blood mononuclear cells of a healthy male donor Mathieu Fieldes, Engi Ahmed, Chloé Bourguignon, Joffrey Mianné, Martin Mélanie, Cécile Arnould, Isabelle Vachier, Said Assou, John de Vos, Arnaud

5 En savoir plus

Expression of miR-142-5p in Peripheral Blood Mononuclear Cells from Renal Transplant Patients with Chronic Antibody-Mediated Rejection

Expression of miR-142-5p in Peripheral Blood Mononuclear Cells from Renal Transplant Patients with Chronic Antibody-Mediated Rejection

Abstract In renal transplantation, the unresponsiveness of patients undergoing chronic antibody mediated rejection (CAMR) to classical treatment stress on the need for accurate biomarkers to improve its diagnosis. We aim to determine whether microRNA expression patterns may be associated with a diagnosis of CAMR. We performed expression profiling of miRNAs in peripheral blood mononuclear cells (PBMC) of kidney transplant recipients with CAMR or stable graft function. Among 257 expressed miRNAs, 10 miRNAs associated with CAMR were selected. Among them, miR-142-5p was increased in PBMC and biopsies of patients with CAMR as well as in a rodent model of CAMR. The lack of modulation of miR-142-5p in PBMC of patients with renal failure, suggests that its over-expression in CAMR was associated with immunological disorders rather than renal dysfunction. A ROC curve analysis performed on independent samples showed that miR-142-5p is a potential biomarker of CAMR allowing a very good discrimination of the patients with CAMR (AUC = 0.74; p = 0.0056). Moreover, its expression was decreased in PHA-activated blood cells and was not modulated in PBMC from patients with acute rejection, excluding a non-specific T cell activation expression. The absence of modulation of this miRNA in immunosuppressed patients suggests that its expression was not influenced by treatment. Finally, the analysis of miR-142-5p predicted targets under-expressed in CAMR PBMC in a published microarray dataset revealed an enrichment of immune-related genes. Altogether, these data suggest that miR-142-5p could be used as a biomarker in CAMR and these finding may improve our understanding of chronic rejection mechanisms.
En savoir plus

8 En savoir plus

Antigen-Specific Gene Expression Profiles of Peripheral Blood Mononuclear Cells Do Not Reflect Those of T-Lymphocyte Subsets

Antigen-Specific Gene Expression Profiles of Peripheral Blood Mononuclear Cells Do Not Reflect Those of T-Lymphocyte Subsets

and DNA Array Unit, National Institute on Aging, Bethesda, Maryland 4 Received 23 April 2004/Returned for modification 28 June 2004/Accepted 1 July 2004 Advances in microarray technology have allowed for the monitoring of thousands of genes simultaneously. This technology is of particular interest to immunologists studying infectious diseases, because it provides tremendous potential for investigating host-pathogen interactions at the level of immune gene expression. To date, many studies have focused either on cell lines, where the physiological relevance is questionable, or on mixed cell populations, where the contributions of individual subpopulations are unknown. In the present study, we perform an intrasubject comparison of antigen-stimulated immune gene expression profiles between a mixed population of peripheral blood mononuclear cells (PBMC) and the two predominant cell types found in PBMC, CD4 ⴙ and CD8 ⴙ T lymphocytes. We show that the microarray profiles of CD4 ⴙ and CD8 ⴙ T
En savoir plus

7 En savoir plus

Transcriptome Analysis Describing New Immunity and Defense Genes in Peripheral Blood Mononuclear Cells of Rheumatoid Arthritis Patients

Transcriptome Analysis Describing New Immunity and Defense Genes in Peripheral Blood Mononuclear Cells of Rheumatoid Arthritis Patients

Methodology/Principal Findings: The Illumina Human-6v2 Expression BeadChips were used for a complete genome-wide transcript profiling of peripheral blood mononuclear cells (PBMCs) from 18 RA patients and 15 controls. Differential analysis per gene was performed with one-way analysis of variance (ANOVA) and P values were adjusted to control the False Discovery Rate (FDR,5%). Genes differentially expressed at significant level between patients and controls were analyzed using Gene Ontology (GO) in the PANTHER database to identify biological processes. A differentially expression of 339 Reference Sequence genes (238 down-regulated and 101 up-regulated) between the two groups was observed. We identified a remarkably elevated expression of a spectrum of genes involved in Immunity and Defense in PBMCs of RA patients compared to controls. This result is confirmed by GO analysis, suggesting that these genes could be activated systemically in RA. No significant down-regulated ontology groups were found. Microarray data were validated by real time PCR in a set of nine genes showing a high degree of correlation.
En savoir plus

11 En savoir plus

Characterization of adenosine A 2 receptors of peripheral blood mononuclear cells of patients with fibromuscular dysplasia

Characterization of adenosine A 2 receptors of peripheral blood mononuclear cells of patients with fibromuscular dysplasia

Figure 1 legend: Adenosine plasma level (Panel A) and A 2A - or A 2B -receptor expression (Panel B) in patients. a: Whole FMD patient population; b : FMD without migraine-like headache; c: migraine patients; d: FMD with migraine-like headache; e: hypertensive patients (hypertension); f: controls. Examples of Western blot of A 2A - (Top) and A 2B - (Bottom) receptor production by peripheral blood mononuclear cells.* p<0.01

7 En savoir plus

Modifications of the endosomal compartment in peripheral blood mononuclear cells and fibroblasts from Alzheimer’s disease patients

Modifications of the endosomal compartment in peripheral blood mononuclear cells and fibroblasts from Alzheimer’s disease patients

F Corlier 1,2,3 , I Rivals 4 , J Lagarde 5 , L Hamelin 5 , H Corne 5 , L Dauphinot 1,2,3 , K Ando 1,2,3 , J-C Cossec 1,2,3 , G Fontaine 1,2,3 , G Dorothée 6,7 , C Malaplate-Armand 8,9 , J-L Olivier 8,9 , B Dubois 10 , M Bottlaender 11 , C Duyckaerts 1,2,3,12 , M Sarazin 5,13 , M-C Potier 1,2,3,13 and the Clinical ImaBio3 team 14 Identification of blood-based biomarkers of Alzheimer’s disease (AD) remains a challenge. Neuropathological studies have identi fied enlarged endosomes in post-mortem brains as the earliest cellular change associated to AD. Here the presence of enlarged endosomes was investigated in peripheral blood mononuclear cells from 48 biologically de fined AD patients (25 with mild cognitive impairment and 23 with dementia (AD-D)), and 23 age-matched healthy controls using immunocytochemistry and confocal microscopy. The volume and number of endosomes were not signi ficantly different between AD and controls. However, the percentage of cells containing enlarged endosomes was signi ficantly higher in the AD-D group as compared with controls. Furthermore, endosomal volumes signi ficantly correlated to [C 11 ]PiB cortical index measured by positron emission tomography in the AD group, independently of the APOE genotype, but not to the levels of amyloid-beta, tau and phosphorylated tau measured in the cerebrospinal fluid. Importantly, we confirmed the presence of enlarged endosomes in fibroblasts from six unrelated AD-D patients as compared with five cognitively normal controls. This study is the first, to our knowledge, to report morphological alterations of the endosomal compartment in peripheral cells from AD patients correlated to amyloid load that will now be evaluated as a possible biomarker.
En savoir plus

9 En savoir plus

Effects of chronic morphine and morphine withdrawal on gene expression in rat peripheral blood mononuclear cells.

Effects of chronic morphine and morphine withdrawal on gene expression in rat peripheral blood mononuclear cells.

Chronic morphine treatment alters gene expression in brain structures. There are increasing evidences showing a correlation, in gene expression modulation, between blood cells and brain in psychological troubles. To test whether gene expression regulation in blood cells could be found in drug addiction, we investigated gene expression profiles in peripheral blood mononuclear (PBMC) cells of saline and morphine-treated rats. In rats chronically treated with morphine, the behavioural signs of spontaneous withdrawal were observed and a withdrawal score was determined. This score enabled to select the time points at which the animals displayed the mildest and strongest withdrawal signs (12h and 36h after the last injection). Oligonucleotide arrays were used to assess differential gene expression in the PBMCs and quantitative real-time RT-PCR to validate the modulation of several candidate genes 12h and 36h after the last injection. Among the 812 differentially expressed candidates, several genes (Adcy5, Htr2a) and pathways (Map kinases, G-proteins, integrins) have already been described as modulated in the brain of morphine-treated rats. Sixteen out of the twenty four tested candidates were validated at 12h, some of them showed a sustained modulation at 36h while for most of them the modulation evolved as the withdrawal score increased. This study suggests similarities between the gene expression profile in PBMCs and brain of morphine treated rats. Thus, the searching of correlations between the severity of the withdrawal and the PBMCs gene expression pattern by transcriptional analysis of blood cells could be promising for the study of the mechanisms of addiction.
En savoir plus

13 En savoir plus

Generation of a human induced pluripotent stem cell line (iPSC) from peripheral blood mononuclear cells of a patient with a myasthenic syndrome due to mutation in COLQ

Generation of a human induced pluripotent stem cell line (iPSC) from peripheral blood mononuclear cells of a patient with a myasthenic syndrome due to mutation in COLQ

e Institute for Regenerative Medicine and Biotherapy, INSERM UMR1183, Univ Montpellier, Montpellier, France A B S T R A C T Congenital myasthenic syndromes (CMS) are a class of inherited disorders affecting the neuromuscular junction, a synapse whose activity is essential for movement. CMS with acetylcholinesterase (AChE) deficiency are caused by mutations in COLQ, a collagen that anchors AChE in the synapse. To study the pathophysiological mechanisms of the disease in human cells, we have generated iPSC from a patient’s Peripheral Blood Mononuclear cells (PBMC) by reprogramming these cells using a non-integrative method using Sendai viruses bearing the four Yamanaka factors Oct3/4, Sox2, Klf4, and L-Myc.
En savoir plus

5 En savoir plus

Whipple's Disease: Diagnostic Value of rpoB Gene PCR from Peripheral Blood Mononuclear Cells.

Whipple's Disease: Diagnostic Value of rpoB Gene PCR from Peripheral Blood Mononuclear Cells.

3 Results Among the 20 patients with WD, only six experienced typical clinical symptoms and revealed characteristic PAS- positive macrophages in the duodenal lamina propria and were thus defined as having classical WD (Table 1 , Fig. 1 ). One further patient revealed atypical PAS-positive cells in the duodenal submucosa (Table 1 , patient 14). Although the specificity of the PAS staining of duodenal biopsies as the classical diagnostic method for WD for our cohort was 100%, the sensitivity (38.9%) and negative predictive value (62.1%) was very low. T. whipplei-specific IHC of the duodenum and rpoB gene PCR from duodenal tissue revealed positive results for all included patients with WD and none of the control subjects (Table 1 ). Among 11 patients with atypical WD, nine presented with only a faint positive T. whipplei-specific IHC within the duodenal submucosa (Table 1 , Fig. 2 a, b). Two patients were ini- tially identified via histological analysis of lymph nodes excised to exclude malignant disease (Fig. 2 c, d), and two patients with isolated T. whipplei-induced endocarditis were diagnosed via analysis of cardiac valves (Table 1 , Fig. 2 e, f). In addition, PCR from cerebrospinal fluid was positive for 8 of 15 patients (Table 1 ).
En savoir plus

12 En savoir plus

Peripheral blood mononuclear cells extracts VEGF protein levels and VEGF mRNA: Associations with inflammatory molecules in a healthy population

Peripheral blood mononuclear cells extracts VEGF protein levels and VEGF mRNA: Associations with inflammatory molecules in a healthy population

anticancer therapy [ 76 – 79 ] and is showing positive predictions for future development of new medications. VEGF isoforms are expressed in different quantities, based on the cell type that is producing them. Most cells express VEGF 121 , VEGF 165 (the most abundant) and VEGF 189 . Quantification of VEGF isoforms in our study confirmed the reports from literature, as the most expressed isoform from the PBMCs population was VEGF 165 , followed by VEGF 121 . VEGF 145 was also quantified in our research, although it is more commonly found in cells of placental origin [ 80 , 81 ]. The notion that the identification of VEGF isoforms is important in fundamental biologi- cal and pharmacological studies has arisen the last few years. Specific VEGF isoforms were studied in different types of cancer [ 82 ], cardiovascular disease [ 83 ], kidney disease [ 84 ], auto- immune disease [ 85 ] and many others. Pro-angiogenic VEGFxxx and anti-angiogenic VEGFxxxb splicing variants are particularly interesting; it has been demonstrated that their ratio varies based on different pathology, therefore, determination of pro and anti-angiogenic isoforms could serve as diagnostic tools for such diseases [ 86 – 88 ]. Moreover, the quantifica- tion of pro and anti-angiogenic isoforms was shown to have a predictive value in response to a treatment with bevacizumab [ 89 ]. However, there is no available method that could separate pro and anti-angiogenic isoforms routinely, therefore, most of the studies quantify only pro- angiogenic isoforms. Likewise, in our research, only pro-angiogenic isoforms were detected and associated with specific protein levels. Still, we are aware of the importance of separation and quantification of all VEGF splice variants. We believe that in the future studies, anti-angio- genic isoforms should be routinely quantified and distinguished from pro-angiogenic isoforms for attempting diagnosis of different chronic diseases and prediction of response of anti-VEGF agents (pharmacogenomics).
En savoir plus

23 En savoir plus

Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

Marjorie Monleau 1,2,3,4 , Sophie Bonnel 1,2,3,4 , Thierry Gostan 1,2,3 , Dominique Blanchard 4 , Valérie Courgnaud 1,2,3* and Charles-Henri Lecellier 1,2,3* Abstract Background: microRNAs (miRNAs) play crucial roles in major biological processes and their deregulations are often associated with human malignancies. As such, they represent appealing candidates as targets of innovative therapies. Another interesting aspect of their biology is that they are present in various biological fluids where, advantageously, they appear to be very stable. A plethora of studies have now reported their potential as biomarkers that can be used in diagnosis, prognosis and/or theranostic issues. However, the application of circulating miRNAs in clinical practices still requires the identification of highly efficient, robust and reproducible methods for their isolation from biological samples. In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology.
En savoir plus

12 En savoir plus

Bovine Viral Diarrhoea Virus Induces Apoptosis in Blood Mononuclear Cells by a Mechanism Largely Dependent on Monocytes

Bovine Viral Diarrhoea Virus Induces Apoptosis in Blood Mononuclear Cells by a Mechanism Largely Dependent on Monocytes

In a variety of viral infections, there is increasing evidence that changes observed in lymphoid cell functions are directly associated with the induction of apoptosis or programmed cell death (Razvi & Welsh, 1995). For BVDV, the occurrence of apoptosis has recently been reported with a few cytopathic strains in long-term cultures, including kidney and testis cells (Zhang et al. , 1996 ; Hoff & Donis, 1997) and bone marrow-derived macrophages (Adler et al. , 1997). The purpose of the present study was to examine the induction of apoptotic cell death by BVDV in peripheral blood mononuclear cells (PBMC). We addressed the question whether the non-cytopathic/ cytopathic biotypes of the same BVDV antigenic pair were able to induce apoptosis in bovine PBMC and if so, to identify the cell population(s) involved. The PBMC were isolated from blood of normal healthy cattle by cen trifugation on Ficoll–Hypaque (Pharmacia). The cells (2x10 /ml) were cultured with the virus inoculum at an m.o.i. of 6·3 TCID 50 per cell in complete RPMI 1640
En savoir plus

9 En savoir plus

Peripheral Blood Mononuclear Cell Metabolism Acutely Adapted to Postprandial Transition and Mainly Reflected Metabolic Adipose Tissue Adaptations to a High-Fat Diet in Minipigs

Peripheral Blood Mononuclear Cell Metabolism Acutely Adapted to Postprandial Transition and Mainly Reflected Metabolic Adipose Tissue Adaptations to a High-Fat Diet in Minipigs

didier.remond@inra.fr (D.R.); dominique.dardevet@inra.fr (D.D.); isabelle.savary-auzeloux@inra.fr (I.S.-A.) * Correspondence: sergio.polakof@inra.fr; Tel.: +33-(0)-473-624-895; Fax: +33-(0)-473-624-638 Received: 29 September 2018; Accepted: 15 November 2018; Published: 21 November 2018   Abstract: Although peripheral blood mononuclear cells (PBMCs) are widely used as a valuable tool able to provide biomarkers of health and diseases, little is known about PBMC functional (biochemistry-based) metabolism, particularly following short-term nutritional challenges. In the present study, the metabolic capacity of minipig PBMCs to respond to nutritional challenges was explored at the biochemical and molecular levels. The changes observed in enzyme activities following a control test meal revealed that PBMC metabolism is highly reactive to the arrival of nutrients and hormones in the circulation. The consumption, for the first time, of a high fat–high sucrose (HFHS) meal delayed or sharply reduced most of the observed postprandial metabolic features. In a second experiment, minipigs were subjected to two-month HFHS feeding. The time-course follow-up of metabolic changes in PBMCs showed that most of the adaptations to the new diet took place during the first week. By comparing metabolic (biochemical and molecular) PMBC profiles to those of the liver, skeletal muscle, and adipose tissue, we concluded that although PBMCs conserved common features with all of them, their response to the HFHS diet was closely related to that of the adipose tissue. As a whole, our results show that PBMC metabolism, particularly during short-term (postprandial) challenges, could be used to evaluate the whole-body metabolic status of an individual. This could be particularly interesting for early diagnosis of metabolic disease installation, when fasting clinical analyses fail to diagnose the path towards the pathology.
En savoir plus

21 En savoir plus

Deformability of Tumor Cells versus Blood Cells

Deformability of Tumor Cells versus Blood Cells

cells that naturally live and survive in circulation versus ones that typically live within a tissue parenchyma, we first compared the passage time and buoyant mass properties of tumor cell lines versus blood cells. Figure 3 shows distinct size and deformability characteristics between these two types of cells (top two rows versus bottom two rows of Fig. 3A,B), where the small differences between mesenchymal and epithelial tumor cell phenotypes pale in comparison to the dramatic differences between the tumor cell types and blood cells. Indeed, blood cells have passage times on the order of a few milliseconds, whereas tumor cells can have passage times as long as a few sec- onds. Included in the various tumor cell lines measured are a lung cancer cell line (H1975), two breast cancer lines (MDA-MB231 and SKBR-3), and a prostate cancer line (PC3-9). Meanwhile, the types of blood cells measured include human erythrocytes, peripheral blood mononuclear cells (PBMC), polymorphonuclear (PMN) leuko- cytes, L1210 (mouse lymphoblast cell line), and primary mouse (BALB/c) leukocytes (Supplementary Methods). As is already known, the majority of blood cells are smaller than most of the tumor cells in culture. However, even for cells of comparable size, as indicated by their buoyant mass, blood cells have decidedly faster passage times than do tumor cells. This distinction holds true for passage time measurements at two different flow rates, as portrayed by the similarity between Fig. 3A,B. For each flow condition, agglomerative clustering was employed on the median of the log 10 values of the passage times for binned buoyant masses, given a set range of buoyant
En savoir plus

12 En savoir plus

Cryopreservation of mononuclear cells before extracorporeal photochemotherapy does not impair their anti-proliferative capabilities

Cryopreservation of mononuclear cells before extracorporeal photochemotherapy does not impair their anti-proliferative capabilities

Universit é d ’ Auvergne, Clermont-Ferrand, France, and 6 EFS Rh ô ne-Alpes, La Tronche, France Abstract Background aims. The clinical benefi ts of extracorporeal photochemotherapy (ECP) are well recognized, but its clinical use is limited by logistical diffi culties, especially because of the need to perform repeated aphereses. The cryopreservation of mononuclear cells could allow maintenance of the ECP schedule while reducing the number of aphereses. The aim of this work was to assess whether previous cryopreservation impairs the immunomodulatory function of ECP-treated peripheral blood mononuclear cells (PBMC). Methods. Fresh or previously cryopreserved PBMC were exposed to ECP and added on day 0 into a mixed leukocyte reaction. Proliferation of alloreactive lymphocytes was measured by carboxyfl uorescein succinimidyl ester (CFSE) dye dilution. Apoptosis was quantifi ed by annexin – 7AAD staining. Results . ECP-induced apop- tosis was slightly increased in cryopreserved cells but the kinetics of apoptosis were similar to fresh cells. Lymphocytes stimulated in the presence of ECP-treated PBMC displayed a signifi cant decrease in proliferation. The suppression was enforced when ECP-treated cells had been activated previously by allogeneic stimulation. Cryopreservation before ECP exposure did not impact apoptosis triggering or anti-proliferative properties of ECP-treated cells. Conclusions. Cryopreser- vation before ECP does not impair the immunomodulatory effects of treated cells. These data warrant investigation of the clinical use of cryopreserved PBMC for ECP.
En savoir plus

9 En savoir plus

Show all 3994 documents...