Presence of Bcr-Abl Positive Cells in Peripheral Blood Leukocytes of Immunosuppressed Solid Organ Transplant (SOT) Recipients

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 3191-3191
Author(s):  
Philipp D. le Coutre ◽  
Petra Reinke ◽  
Ruth Neuhaus ◽  
Ralf Trappe ◽  
Frauke Ringel ◽  
...  
Keyword(s):  
Peripheral Blood ◽  
Case Reports ◽  
Control Group ◽  
Study Group ◽  
Solid Organ ◽  
Healthy Individuals ◽  
Peripheral Blood Leukocytes ◽  
Rt Pcr ◽  
Blood Leukocytes ◽  
Female Ratio

Abstract In chronic myeloid leukemia (CML) the p210 BCR-ABL protein, generated by a t(9;22)(q34;q11) translocation, is the underlying mechanism of leukemogenesis. Although the presence of p210 BCR-ABL is normally restricted to CML patients (pts), previous reports demonstrated low levels of bcr-abl transcripts in healthy individuals that may be controlled by an intact immune system (Bose et al. Blood92:3362, 1998). Interestingly, several articles reported cases of CML occurring in pts after SOT (Pelloso et al. Leukemia Res29:353, 2005). Therefore, immunosuppressed SOT recipients should represent an optimal population to investigate the frequency of bcr-abl transcripts in non-leukemic individuals in order to address a potential impact of immunosuppression on the presence of bcr-abl transcripts. This possibility was investigated by studying peripheral blood leukocytes for the presence of bcr-abl transcripts in a total of 201 individuals of whom 100 were SOT recipients (n=50 kidney, n=46 liver, n=3 heart, n=1 heart-lung) and 101 were control individuals (n=87 patients with renal failure, n=14 healthy individuals). The male to female ratio in the study group was 54:46 (median age: 55.38 years, range: 22–83) and matched the control group (median age: 59.8 years, range: 32–96). Included in the study group were 13 pts with post transplant malignancies who were previously treated with standard chemotherapeutic regimens. Immune suppressive drugs given to all SOT recipients included steroids (90%), cyclosporine A (69%), azathioprine (22%), tacrolimus (69%), sirolimus (44%), mycophenolic acid (63%) and others (37%). For the detection of the bcr-abl transcript we used a nested reverse transcriptase-polymerase chain reaction (RT-PCR) assay that is routinely used in our institution. All samples were tested at least twice. In 5/100 immunosuppressed pts at least 1 of 2 RT-PCR products was bcr-abl positive in the second round amplification. This rate significantly exceeded the control group that was completely bcr-abl negative (0/101 p = 0.0242). Of the 5 bcr-abl positive pts, 3 were liver transplant and 2 were kidney transplant recipients. The latency in this group (interval between transplantation and bcr-abl PCR) was at 58.4 months (range: 24-135 months) and shorter when compared to the latency in the remaining study group (104.8 months; range 1 – 369). Additionally, all samples were tested for both the pml-rara and aml1-eto transcripts that are detectable in subsets of pts with acute myeloid leukaemia but none of the samples tested positive for these transcripts. Our findings are extended by three case reports of SOT recipients (2 kidney, 1 liver) who developed CML in a total of 2088 transplantations in our centre in 9 years. In these three pts (2 males, 1 female), CML occurred at 84.3 months (range: 32–183) following SOT. In summary our data show: 1. The presence of bcr-abl transcripts in immunosuppressed non-leukemic SOT pts and the absence of bcr-abl transcripts in healthy normals when tested with routine PCR protocols. 2. The absence of pml-rara and aml1-eto transcripts in both the study and control group. And 3. the occurrence of CML in three patients following SOT suggesting a higher frequency of CML in this population.

scholarly journals Hyperexpression of TLR2 and TLR4 in patients with ischemic stroke in acute period of the disease

2020 ◽  
Vol 22 (4) ◽  
pp. 665-674
Author(s):  
L. V. Gankovskaya ◽  
L. V. Stakhovskaya ◽  
V. V. Grechenko ◽  
E. A. Koltsova ◽  
O. S. Uvarova ◽  
...  
Keyword(s):  
Gene Expression ◽  
Innate Immunity ◽  
Ischemic Stroke ◽  
Peripheral Blood ◽  
Surface Expression ◽  
Control Group ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Healthy Donors ◽  
Tlr4 Gene

Pathogenesis of ischemic stroke  is actively  involved  in the  system  of innate immunity. Under conditions of cerebral  ischemia, a number of biologically  active  substances are  released  that  interact with innate immunity receptors, in particular TLR2  and  TLR4, which  exacerbate inflammation in brain  tissue. Identification of predictor markers  at the level of the innate immunity system may foresee the clinical course of ischemic stroke and ensure timely treatment. Our objective was to study expression of TLR2 and TLR4 receptors in peripheral blood leukocytes  in patients with ischemic stroke in the dynamics of the disease. 27 people  were included in the study. The main  group consisted of patients with ischemic stroke of varying severity (n = 19). Patients of the main  group were divided into two subgroups:  with an NIHSS index value of < 10 (n = 10) and > 10 (n = 9). The control group included healthy  donors  with no history  of acute  and chronic inflammatory diseases (n = 8). Peripheral blood  leukocytes  were used as the  test material. To determine expression  of the TLR2  and TLR4  genes, RT-PCR in real time was used. Surface  expression  of TLRs was determined by flow cytometry. A study of the TLR2 and TLR4 gene expression showed that on the 1st, 3rd  and 7th  day post-stroke, the TLR4 gene expression  in patients was significantly  increased, when compared to the control group (p < 0.01), whereas TLR2 gene expression on the 3rd  day of the disease was not statistically different from the control group. A study of surface expression  of receptors showed that the average TLR2 fluorescence intensity on the patients’ peripheral blood monocytes was significantly  increased on the 1st  and 3rd  day of disease when compared to the control group.  The  surface  expression  of TLR4  on monocytes has a statistically significant  increase  only on day 7. Assessment  of surface expression  of TLRs in subgroups  with different  severity values by NIHSS showed that  patients with a NIHSS index > 10 had a significantly  higher  level of surface of TLR2  expression  over the observation period, while the largest difference in TLR4  expression  in the subgroups  was observed  on the 1st day of the disease (p < 0.05). Patients with ischemic stroke showed an increase  in TLR2 and TLR4 expression at the gene and protein level, compared to healthy  donors. These indices can be considered possible predictors for clinical  prognosis  of ischemic stroke.

scholarly journals Thymic Microchimerism Correlates with the Outcome of Tolerance-Inducing Protocols for Solid Organ Transplantation

2001 ◽  
Vol 12 (12) ◽  
pp. 2815-2826
Author(s):  
Marina Noris ◽  
Daniela Cugini ◽  
Federica Casiraghi ◽  
Nadia Azzollini ◽  
Luciana De Deus Viera Moraes ◽  
...  
Keyword(s):  
Graft Survival ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Solid Organ Transplantation ◽  
Pcr Analysis ◽  
Solid Organ ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Peripheral Blood Mononuclear

ABSTRACT. This study found that pretransplant infusion of donor peripheral blood leukocytes, either total leukocytes (peripheral blood leukocytes) or peripheral blood mononuclear cells (PBMC), under appropriate immunomodulating conditions was more effective than donor bone marrow (BM) in prolonging the survival of rats that received kidney grafts. A higher percentage of MHCII+ cells was found in donor PBMC than in BM cells, and depletion of MHCII+ cells from donor PBMC abolished their tolerogenic potential. By the analysis of microchimerism in rats infused with donor cells and killed at different time points thereafter, the better tolerogenic potential of leukocyte infusion related to a higher capability of these cells to engraft the recipient thymus. PCR analysis on OX6-immunopurified cells revealed the presence of donor MHCII+ cells in the thymus of these animals. The role of intrathymic microchimerism was reinforced by findings that thymectomy at the time of transplant prevented tolerance induction by donor leukocytes. Donor DNA was found in the thymus of most long-term graft animals that survived, but in none of those that rejected their grafts. The presence of intrathymic microchimerism correlated with graft survival, and microchimerism in other tissues was irrelevant. PCR analysis of DNA from thymic cell subpopulations revealed the presence of donor MHCII+ cells in the thymus of long-term surviving animals. Thus, in rats, donor leukocyte infusion is better than donor BM for inducing graft tolerance, defined by long-term graft survival, donor-specific T cell hyporesponsiveness, and reduced interferon gamma production. This effect appears to occur through migration of donor MHCII+ cells in the host thymus.

scholarly journals Association of FOXP3 gene -3279 C>A polymorphism with the risk of pulmonary sarcoidosis

2017 ◽  
Vol 89 (12) ◽  
pp. 64-67 ◽  
Author(s):  
I E Malysheva ◽  
L V Topchieva ◽  
E L Tikhonovich ◽  
I V Kurbatova ◽  
O V Balan
Keyword(s):  
Peripheral Blood ◽  
Polymorphic Marker ◽  
Pulmonary Sarcoidosis ◽  
Control Group ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Foxp3 Gene ◽  
Healthy Donors ◽  
Gene Transcripts ◽  
The Republic

Aim. To investigate the association of the polymorphic marker -3279 C>A of the FOXP3 gene with the risk of pulmonary sarcoidosis (PS) and to estimate the transcription level of this gene in the carriers of different genotypes of this polymorphic marker. Subjects and methods. The investigation included 99 patients of Russian ethnicity (mean age, 45.41±1.31 years) living in the Republic of Karelia, who were diagnosed with persistent PS, and 116 healthy donors (mean age, 42.06±1.30 years) in the control group. The alleles and genotypes of the polymorphic marker -3279 C>A of the FOXP3 gene were identified using polymerase chain reaction (PCR)-restriction fragment length polymorphism. The number of transcripts of the studied gene in the peripheral blood leukocytes of healthy donors and PS patients was determined with real-time PCR. Results. The control group and the PS patient one had no statistically significant differences in the distribution of the frequencies of alleles and genotypes by the polymorphic marker –308G>A of the FOXP3 gene (p > 0.05). The number of FOXP3 gene transcripts was not statistically significantly different in the peripheral blood leukocytes of patients with PS and control individuals. No statistically significant differences were observed in the mRNA expression levels in the above-mentioned gene in the carriers of different genotypes by the polymorphic marker -3279 C>A of the FOXP3 gene in all examined groups. Conclusion. The polymorphic marker -3279 C>A of the FOXP3 gene is unassociated with the risk of PS.

scholarly journals Genetic Polymorphisms of Metabolic Enzyme Genes Associated with Leukocyte Mitochondrial DNA Copy Number in PAHs Exposure Workers

2020 ◽  
Author(s):  
Xinling Li ◽  
Xiaoran Duan ◽  
Hui Zhang ◽  
Mingcui Ding ◽  
Yanbin Wang ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Genetic Polymorphisms ◽  
Peripheral Blood ◽  
Quantitative Polymerase Chain Reaction ◽  
Metabolic Enzymes ◽  
Control Group ◽  
Peripheral Blood Leukocytes ◽  
Chain Reaction ◽  
Blood Leukocytes ◽  
Polymerase Chain

Abstract Background: PAHs exposure had been reported to be a risk factor of mtDNAcn in our early study. However, the effect of metabolic enzymes’ genetic polymorphisms on mtDNAcn in PAHs-Exposure workers has not been fully evaluated.Methods: We investigated the effects of metabolic enzymes’ genetic polymorphisms on mtDNAcn among 544 coke oven workers and 238 office staffs. The mtDNAcn of peripheral blood leukocytes was measured using Real-time quantitative polymerase chain reaction method. Polymerase chain reaction and restriction fragment length was used to detect five polymorphisms in GSTT1, GSTM1, GSTP1 rs1695, CYP2E1 rs6413432, and CYP2E1 rs3813867.Results: The mtDNAcn in peripheral blood leukocytes was significantly lower in the exposure group than that in the control group (P < 0.001). The 1-OHPYR had an increasing trend with the genotypes AA→AG→GG of GSTP1 rs1695 in the control group. Generalized linear model indicated that the influencing factors of mtDNAcn were PAHs-exposure [b(95% CI) = -0.420 (-0.469, -0.372), P < 0.001], male [β(95% CI) = -0.058 (-0.103, -0.012), P = 0.013] ,and AA genotype for GSTP1 rs1695 [β(95% CI) = -0.051 (-0.095, -0.008), P = 0.020].Conclusions: The male was susceptibility to PAHs exposure. The AA genotype of GSTP1 rs1695 may influence the toxicity of PAHs and associated with the decreased of mtDNAcn.

scholarly journals Deoxyribonucleic acid methylation in the enhancer region of the CDKN2A/2B and CDKN2B-AS1 genes in blood vessels and cells in patients with carotid atherosclerosis

2020 ◽  
Vol 25 (10) ◽  
pp. 4060
Author(s):  
Yu. A. Koroleva ◽  
A. V. Markov ◽  
I. A. Goncharova ◽  
A. A. Sleptsov ◽  
N. P. Babushkina ◽  
...  
Keyword(s):  
Dna Methylation ◽  
Peripheral Blood ◽  
Methylation Level ◽  
Deoxyribonucleic Acid ◽  
Healthy Individuals ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Blood Samples ◽  
Cpg Sites ◽  
Atherosclerotic Lesions

Aim. Comparative analysis of the deoxyribonucleic acid (DNA) methylation level in the enhancer region of the CDKN2A/2B and CDKN2B-AS1 genes (9p21.3 locus) in vessels with/without atherosclerotic lesions, as well as in leukocytes of patients with clinically relevant carotid artery (CA) atherosclerosis and healthy individuals.Material and methods. The group of patients with clinically relevant atherosclerosis included 22 individuals with severe stenosis (>80%) of CA. Samples of atherosclerotic plaques, presenting CA regions, and great saphenous veins, as well as peripheral blood samples (leukocytes) were obtained from patients. The control group consisted of 14 individuals with the mild CA stenosis (£24%) and without hemodynamically relevant changes; peripheral blood samples were obtained from each of them. DNA methylation level was assessed by targeted bisulfite sequencing of amplicons.Results. The tissue-specific methylation of 31 CpG-site in the CDKN2A/2B and CDKN2B-AS1 gene enhancer was established: the vascular tissues significantly differed from the peripheral blood leukocytes. At the same time, there was an increase in the methylation level of both certain CpG sites and whole analyzed CA region affected by atherosclerosis (48,6 [34,8; 62,0]%), compared with intact vessels, both arteries (25,2 [23,1; 41,60]%, p=0,0001) and veins (35,0 [31,6; 40,0]%, p=0,0039). Patients had lower methylation levels in all CpG sites in blood leukocytes compared to blood vessel samples (8,7 [6,1; 9,7]%; p<0,05). At the same time, the level of DNA methylation in the blood leukocytes of atherosclerotic patients does not differ from that in healthy individuals (9,3 [8,3; 13,6]%; p>0,8).Conclusion. In the present study, the relationship between an increase in the DNA methylation in the enhancer of the CDKN2A/2B and CDKN2B-AS1 genes in CA and their atherosclerotic lesions was revealed, as well as the tissue-specific DNA methylation between vessels and peripheral blood leukocytes.

scholarly journals LPL methylation of peripheral blood leukocytes - Prophet of atherothrombotic stroke

2020 ◽  
Author(s):  
Minjia Xiao ◽  
Zhijie Xiao
Keyword(s):  
Ischemic Stroke ◽  
Peripheral Blood ◽  
Methylation Level ◽  
Epigenetic Modification ◽  
Hunan Province ◽  
Control Group ◽  
P Value ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
Cpg Dinucleotides

Abstract Background: Lipoprotein lipase (LPL) is a glycoprotein enzyme playing a pivotal role in energy and lipoprotein metabolism. It hydrolyzes triglyceride-rich lipoprotein and produces fatty acids as well as monoacylglycerol. Multiple risk factors including age, sex, hypertension, smoking and hyperlipidemia contribute to ischemic stroke (IS). Hyperlipidemia is an extremely pivotal risk factor for ischemic stroke especially atherothrombotic stroke (ATS) in that it can lead to intracranial or extracranial vessels’ atherosclerosis analogous to atherosclerosis of coronary artery. However, potential epigenetic mechanisms contributing to IS are not been explored thoroughly. The aim of this study was to illuminate relationship among individuals’ peripheral blood leukocytes methylation level of LPL-promoter-CpG dinucleotides, serum lipid and ATS of Chinese Han population in Hunan Province. Methods: Peripheral blood samples were collected from acute atherothrombotic stroke patients and healthy people, and methylation level of cytosine–phosphate–guanine (CpG) island in LPL promoter region was measured by qPCR and pyrosequencing. Biochemical and anthropometric variables have also been taken into consideration.Results: Integral LPL-promoter-CpG dinucleotides methylation level of case group is evidently higher than control group (38.8±8.4% percentile vs 25.7±6.6% percentile, P value=0.000). DNA methylation at the CpG9~16 locus and CpG20~21 locus was related to ATS after adjusting for gender, previous history of diabetes and hypertension, smoking, drinking, body mass index, and blood lipid levels(CpG9 49.3±24.9, 31.3±13.6, P value =0.02; CpG10 61.3±24.5, 34.0±18.4, P value =0.002; CpG11 71.3±17.3, 32.0±21.1, P value =0.000; CpG12 75.3±17.7, 44.7±19.6, P value =0.000; CpG13 72.0±20.4, 50.0±25.4, P value =0.014; CpG14 63.3±18.0, 45.3±22.0, P value =0.021; CpG15 56.7±13.5, 34.7±14.6, P value =0.000; CpG16 50.0±12.5, 31.3±20.0, P value =0.005; CpG20 52.0±13.2, 27.3±20.9, P value =0.001; CpG21 55.3±11.9, 34.0±22.3, P value =0.004). There is no statistically significant connection between either carotid atherosclerosis or gender and methylation level. Besides, we found a negative association between LPL methylation status and HDL-C levels, whereas LPL gene methylation was linked with LDL-C levels and age positively.Conclusion: We illustrate that epigenetic modification of LPL methylation may have an influence on lipids metabolism and occurrence of ATS, which may be a promising indicator for occurrence risk of ischemic stroke. Whether these findings are valid need further warrant and future prospective.

scholarly journals Analysis of an association of TNF –308G>A polymorphism with a risk for pulmonary sarcoidosis in the Russian population of the Republic of Karelia

2017 ◽  
Vol 89 (3) ◽  
pp. 61-64
Author(s):  
I E Malysheva ◽  
L V Topchieva ◽  
E L Tikhonovich ◽  
O Yu Barysheva
Keyword(s):  
Peripheral Blood ◽  
Inflammatory Responses ◽  
Clinical Manifestations ◽  
Pulmonary Sarcoidosis ◽  
Russian Population ◽  
Control Group ◽  
Mrna Levels ◽  
Peripheral Blood Leukocytes ◽  
Blood Leukocytes ◽  
The Republic

Aim. To analyze an association of TNF –308G>A polymorphism with a risk for pulmonary sarcoidosis (PS) in the Russian population of the Republic of Karelia Subjects and methods. 84 patients with persistent PS and 96 donors without clinical manifestations of this disease (a control group) were examined. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis was used to identify alleles and genotypes by the marker of TNF –308G>A polymorphism. The level of transcripts of the above gene in the peripheral blood leukocytes of healthy and sick people was determined by real-time PCR. Results. There were no significant differences in the distribution of allelic and genotypic frequencies by the marker of TNF –308G>A polymorphism between the control and PS patient groups. There was a significant increase in the number of TNF gene transcripts in the peripheral blood leukocytes of patients with PS compared to the controls. At the same time, there were no marked differences in mRNA expression levels in the above gene in the carriers of different genotypes by the marker of TNF –308G>A polymorphism in all the examined groups. Conclusion. The marker of TNF –308G>A polymorphism is unassociated with the risk of PS in the Russian population of the Republic of Karelia. No differences in TNF mRNA levels in the carriers of different genotypes by the above marker may suggest that the found elevated level of transcripts in the above gene in patients with diagnosed with PS is due to the development of the body’s inflammatory responses in this disease.

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