Abstract 3354: Increase of Telomere Gap between Granulocytes and Lymphocytes in Patients with Previous Myocardial Infarction: A Novel Measure for Accelerated Telomere Erosion

Individuals with age-corrected reduction of total leukocyte telomere length have been shown to exhibit a remarkable increase in mortality from cardiovascular and infectious complications. Given the considerable interindividual variability of telomere length in healthy adults due to genetic differences already present at birth and the heterogeneity in composition of the total leukocyte compartment, extensive study populations are required to detect accelerated telomere shortening between age-matched groups. With this study, we intended to identify a parameter to improve prediction of individual telomere erosion in patients (pts) with coronary artery disease and previous myocardial infarction (CAD). Methods: Mean telomere length (mTL) was measured in 63 CAD pts (mean age 65±4 years, mean ejection fraction 31±9%) and 23 age-matched healthy controls (65±4 years). Using the flow-FISH method, we were able to distinguish between myeloid and lymphoid cell populations in peripheral blood cells as well as in bone marrow mononuclear cells (BMC). Results: The difference between mTL of the granulocyte and lymphocyte population (ΔTEL Gr-Ly ) was significantly increased from 742 ± 396 base pairs (bp) in the control group to 1055±570 bp in CAD pts (p=0.018). Age was the only independent predictor for telomere length of BMCs. Lymphocytes, but not granulocytes, demonstrated significant telomere shortening between BMCs and peripheral blood in CAD patients (−444 ± 624 bp) as opposed to an increase in a young and healthy control group (+307 ± 372 bp) (p<0.001). Using immunomagnetic cell sorting, telomerase activity (TA) was determined in freshly isolated white blood cell subpopulations. Surprisingly, TA could not be detected in peripheral blood granulocytes (CD15 + granulocytes) from CAD pts. In contrast, CD4 + T-lymphocytes contained the highest amount of TA, compared to CD8 + cells and B-lymphocytes. Conclusions: Our results suggest that primarily peripheral blood lymphocytes - and not granulocytes - can develop stress-induced telomere erosion in CAD. Therefore the base pair length difference between the two populations provides a valuable parameter to detect telomere erosion in patients with CAD, largely independent of the hereditary influence on mTL.

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Screening of Telomere Length in Peripheral Blood Lymphocytes From Patients with Aplastic Anemia by Flow-Fluorescence in Situ Hybridization.

Blood ◽

10.1182/blood.v114.22.3205.3205 ◽

2009 ◽

Vol 114 (22) ◽

pp. 3205-3205

Author(s):

Nobuhiro Nishio ◽

Yoshiyuki Takahashi ◽

Hideki Muramatsu ◽

Asahito Hama ◽

Toshihito Nagata ◽

...

Keyword(s):

In Situ Hybridization ◽

Telomere Length ◽

Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Telomere Shortening ◽

Blood Lymphocytes ◽

Normal Individuals ◽

Telomere Signal ◽

Normal Controls

Abstract Abstract 3205 Poster Board III-142 Telomeres in peripheral blood cells from patients with aplastic anemia (AA) are shorter than those from normal individuals. One possible explanation is that telomere shortening reflects the extent and duration of damage to hematopoietic stem cells and the number of compensatory stem cell divisions. Patients with such stressed hematopoiesis probably do not respond to immunosuppressive therapy (IST). Cryptic forms of dyskeratosis congenita (DC) have been recognized in patients with AA who do not have physical abnormalities, but who harbor the gene mutation associated with telomere maintenance. To identify such patients is important because those who have these mutations might respond poorly to IST. The present study evaluates the usefulness of screening telomere length in peripheral blood lymphocytes from patients at the time of diagnosis with AA by flow-fluorescence in situ hybridization (flow-FISH) using Telomere PNA kits (Dako Cytomation, Glostrup, Denmark). After gating diploid cells based on propidium iodine staining, lymphocytes were isolated according to size and granularity. Relative telomere length (RTL) was calculated as the ratio between the telomere signal of each sample and the telomere signal of the control cell line. Because telomeres shorten with age, we obtained age-adjusted measurements for comparison. We used flow-FISH to measure the length of telomeres in peripheral blood lymphocytes from five and two patients with classical and cryptic DC who had the TERT mutation, respectively, to determine whether these two forms of DC can be screened. Significant telomere shortening was detected in all seven patients compared with age-matched normal individuals. To determine the association between telomere length and response to IST, we compared telomere lengths in peripheral blood lymphocytes from 61 normal controls and 27 patients with AA (very severe, severe and non-severe, n = 9 each) at diagnosis who received IST with antithymocyte globulin (ATG) and cyclosporine. The median age of the patients was 9.0 years (range, 1.5 to 14.8). The causes of AA were unknown etiology in 25 patients and hepatitis in 2. In total, 17 of the 27 patients (63%) responded to IST at 6 months after administration of ATG. Age-adjusted delta RTLs did not significantly differ between normal controls and responders (p = 0.181), but those in non-responders were significantly lower than those in normal individuals (p = 0.000). Our findings suggest that screening telomere length in patients with AA at diagnosis is useful for detecting cryptic DC and for predicting responses to IST. Disclosures No relevant conflicts of interest to declare.

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Autologous Peripheral Blood Mononuclear Cells for Limb Salvage in Diabetic Foot Patients with No-Option Critical Limb Ischemia

Journal of Clinical Medicine ◽

10.3390/jcm10102213 ◽

2021 ◽

Vol 10 (10) ◽

pp. 2213

Author(s):

Alessia Scatena ◽

Pasquale Petruzzi ◽

Filippo Maioli ◽

Francesca Lucaroni ◽

Cristina Ambrosone ◽

...

Keyword(s):

Peripheral Blood Mononuclear Cells ◽

Critical Limb Ischemia ◽

Diabetic Foot ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Limb Ischemia ◽

Control Group ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Peripheral blood mononuclear cells (PBMNCs) are reported to prevent major amputation and healing in no-option critical limb ischemia (NO-CLI). The aim of this study is to evaluate PBMNC treatment in comparison to standard treatment in NO-CLI patients with diabetic foot ulcers (DFUs). The study included 76 NO-CLI patients admitted to our centers because of CLI with DFUs. All patients were treated with the same standard care (control group), but 38 patients were also treated with autologous PBMNC implants. Major amputations, overall mortality, and number of healed patients were evaluated as the primary endpoint. Only 4 out 38 amputations (10.5%) were observed in the PBMNC group, while 15 out of 38 amputations (39.5%) were recorded in the control group (p = 0.0037). The Kaplan–Meier curves and the log-rank test results showed a significantly lower amputation rate in the PBMNCs group vs. the control group (p = 0.000). At two years follow-up, nearly 80% of the PBMNCs group was still alive vs. only 20% of the control group (p = 0.000). In the PBMNC group, 33 patients healed (86.6%) while only one patient healed in the control group (p = 0.000). PBMNCs showed a positive clinical outcome at two years follow-up in patients with DFUs and NO-CLI, significantly reducing the amputation rate and improving survival and wound healing. According to our study results, intramuscular and peri-lesional injection of autologous PBMNCs could prevent amputations in NO-CLI diabetic patients.

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Acute Myocardial Infarction Enhances Production of MMP-9 by Peripheral Blood Mononuclear Cells

Heart Lung and Circulation ◽

10.1016/j.hlc.2009.05.464 ◽

2009 ◽

Vol 18 ◽

pp. S306 ◽

Cited By ~ 1

Author(s):

Lu Fang ◽

Xiao-Jun Du ◽

Xiao-Ming Gao ◽

Anthony Dart

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

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Abstract 239: Stem Cell Therapy Improves Critical Limb Ischemia

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.32.suppl_1.a239 ◽

2012 ◽

Vol 32 (suppl_1) ◽

Author(s):

Alaa Marzouk

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Stem Cell ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Embryonic Stem ◽

Limb Ischemia ◽

Control Group ◽

Chronic Limb Ischemia

Introduction: The journey from single cell to complex being is attributable to stem cells role. Adult stem cells originate during ontogeny & persist in specialized niches within organs. Asymmetric division of each stem cell during differentiation produces : one daughter stem cell & one daughter transit amplifying/intermediate cell having migratory properties. Forced migration of hematopoietic stem/progenitor cells (HSPC) from bone marrow into peripheral blood is called mobilization. Accumulating evidence suggests that attenuation of the chemokine stromal derived factor-1(SDF-1)-CXCR4 axis that plays a pivotal role in retention of HSPC in bone marrow (BM) results in the release of these cells from the BM into peripheral blood. Recently, adult cells have been genetically reprogrammed to an embryonic stem cell like state. Induced pluripotent stem cells (IPSCs) were similar to human embryonic stem cells in morphology, proliferative capacity, expression of cell surface antigens, & gene expression. Treatment of ischemic vascular disease of lower limbs remains a significant challenge. Unfortunately, if medical & surgical salvage procedures fail, amputation is an unavoidable result for those patients. Aim of Work: (Hypothesis) To assess the application of implantation of autologous stem/progenitor cell in the treatment of chronic limb ischemia & to evaluate the safety, efficacy & feasibility of this novel therapeutic approach. Methods: A total of 24 patients with chronic limb ischemia not eligible for arterial reconstruction or endovascular procedures were enrolled & randomized (1:1) to either the implanted group or the control group. Control group: Conventional medical therapy in the form of anti platelet therapy & vasodilators. Implanted group: Subcutaneous injection of 300μ g/day of recombinant human granulocyte colony stimulating factor (G-CSF) for 5 days to mobilize stem/progenitor cells from BM. Total leucocytic count is measured daily to follow up successful mobilization of bone marrow mononuclear cells (BMMNCs). Stem cell Harvesting After 5 days peripheral blood mononuclear cells (PBMNCs) were harvested using a cell separator. Samples from apheresis products are subjected to TLC measurement & immunophenotypic characterization of CD34+ cells by flow cytometry. The collected PBMNCs were implanted by multiple intramuscular injections into ischemic limbs. Results: There was significant increase in pain free walking distance & ankle/brachial index (ABI) & significant decreased rest pain. Effectiveness was documented by : reduced number of amputation, increase ABI & improvement of the quality of life in therapeutic group compared to control group. Conclusion: The novel therapeutic approach of PBMNCs implantation in patients with chronic limb ischemia is safe, feasible & effective in decreasing co-morbidity & rate of amputation. Safety was manifested by absence of complications during G-CSF therapy or during harvesting & injection of the stem cells. Recommendations: 1- Future studies on larger number of patients & longer follow up. 2- Controlled studies using different methods & different cell population (PBMNCs, BMMNCs or MSCs) to compare the outcome of each. 3-Studing the role of endothelial progenitor cell dysfunction in different ischemic diseases to develop successful gene therapy.

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Toll like receptor 9 (TLR 9) expression in peripheral blood mononuclear cell (PBMCs) from treated and untreated Grave’s disease patients

QJM ◽

10.1093/qjmed/hcab100.083 ◽

2021 ◽

Vol 114 (Supplement_1) ◽

Author(s):

Ashraf M Okba ◽

Rasha Y Shaheen ◽

Gehan M. H Mostafa ◽

Hanan M Ali ◽

Sylvia W Abo El Fadle ◽

...

Keyword(s):

Peripheral Blood Mononuclear Cell ◽

Mononuclear Cell ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Peripheral Blood Lymphocytes ◽

Graves Disease ◽

Toll Like Receptor ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Abstract Background It is well known that Autoimmune thyroid disease is multifactorial with multiple genetic and environmental factors, immune malfunction also incriminated in the development of this disease, The exact pathogenesis of this disease remains unclear despite the fact that the production of autoantibodies destroys self-tolerance and agitate the adaptive immune system. Our study will answer the question is there a difference in Toll like receptor 9 (TLR 9) expression in peripheral blood mononuclear cell (PBMCs) from Grave’s disease patients. Objective to measure TLR9 percentage expression on peripheral blood mononuclear cells in patients with Graves’ disease. Methods 60 subjects were included in this study; 30 with Graves’ disease and 30 healthy individuals as control group. All the patients were subjected to the following: Full history, clinical examination, thyroid functions, Thyroid ultrasound, Radioisotope thyroid scan: to assess uptake of thyroid gland and Toll like receptor 9 (TLR 9) percentage expression on peripheral blood mononuclear cells will be analyzed using flow cytometry technique. Results The present study proved that patients with Graves’ disease had higher levels of percentage expression of TLR 9 on peripheral blood lymphocytes. Conclusion percentage expression of TLR9 on peripheral blood lymphocytes is higher in Graves’ patients.

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Characteristics of a Pathogenic Molecular Clone of an End-Stage Serum-Derived Variant of Simian Immunodeficiency Virus (SIVF359)

Journal of Virology ◽

10.1128/jvi.75.19.9328-9338.2001 ◽

2001 ◽

Vol 75 (19) ◽

pp. 9328-9338 ◽

Cited By ~ 3

Author(s):

Lennart Holterman ◽

Rob Dubbes ◽

James Mullins ◽

Gerald Learn ◽

Henk Niphuis ◽

...

Keyword(s):

Simian Immunodeficiency Virus ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Human Peripheral Blood ◽

Peripheral Blood Lymphocytes ◽

Peripheral Blood Mononuclear ◽

Molecular Clone ◽

Immunodeficiency Virus ◽

End Stage

ABSTRACT End-stage simian immunodeficiency virus (SIV) isolates are suggested to be the most fit of the evolved virulent variants that precipitate the progression to AIDS. To determine if there were common characteristics of end-stage variants which emerge from accelerated cases of AIDS, a molecular clone was derived directly from serum following in vivo selection of a highly virulent SIV isolate obtained by serial end-stage passage in rhesus monkeys (Macaca mulatta). This dominant variant caused a marked cytopathic effect and replicated to very high levels in activated but not resting peripheral blood lymphocytes. Furthermore, although this clone infected but did not replicate to detectable levels in rhesus monocyte-derived macrophages, these cells were able to transmit infection to autologous T cells upon contact. Interestingly, although at low doses this end-stage variant did not use any of the known coreceptors except CCR5, it was able to infect and replicate in human peripheral blood mononuclear cells homozygous for the Δ32 deletion of CCR5, suggesting the use of a novel coreceptor. It represents the first pathogenic molecular clone of SIV derived from viral RNA in serum and provides evidence that not only the genetic but also the biological characteristics acquired by highly fit late-stage disease variants may be distinct in different hosts.

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Abstract 697: Mobilization of Oct-4 + Bone Marrow-Derived Very Small Embryonic-Like Stem Cells in Patients with Acute Myocardial Infarction

Circulation ◽

10.1161/circ.116.suppl_16.ii_130-a ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Wojciech Wojakowski ◽

Magda Kucia ◽

Boguslaw Machalinski ◽

Edyta Paczkowska ◽

Joanna Ciosek ◽

...

Keyword(s):

Myocardial Infarction ◽

Acute Myocardial Infarction ◽

Stem Cells ◽

Bone Marrow ◽

Pluripotent Stem Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Small Cells ◽

Acute Mi

Bone marrow-derived CD34 + CXCR4 + progenitor cells are mobilized into peripheral blood early in acute myocardial infarction (MI). Adult murine bone marrow contains population of small CD34 + lin − CD45 − CXCR4 + cells expressing markers of pluripotent stem cells (PSC) SSEA, Oct-4 and Nanog. This population of very small embryonic-like cells (VSEL) has unique morphology (small size 2– 4 μm, large nucleus, euchromatin) and capability to form embrioid bodies (EB). Murine EB-derived cells can in vitro differentiate into cells from all three germ layers including cardiomyocytes. We hypothesized that in patients with acute MI small cells expressing the VSEL immunophenotype and PSC markers are present in bone marrow and mobilized into peripheral blood. Blood samples (20 mL) from 18 patients with acute MI were obtained after 12 hours, 2 and 5 days after symptoms onset. Bone marrow samples (20 mL) were obtained from 2 patients with acute MI and 3 healthy volunteers. Mononuclear cells were isolated using hypotonic lysis and samples were analyzed by FACS. Mobilization of following cell populations was confirmed: hematopoietic lin − CD45 + CXCR4 + , lin − CD45 + CD133 + , lin − CD45 + CD34 + and non-hematopoietic (VSEL) lin − CD45 − CXCR4 + , lin − CD45 − CD133 + , lin − CD45 − CD34 + . Analysis of the cell number using lymphocyte gate showed more significant increase of CD45 + (hematopoietic) populations of lin − CD34 + , lin − CD133 + and lin − CXCR4 + cells. After gating for small events (VSEL size range) we found more significant mobilization of small, non-hematopoietic populations of lin − CD34 + , lin − CD133 + and lin − CXCR4 + cells (Table ). The expression of PSC markers (Oct-4, Nanog, SSEA-1) in VSEL was confirmed using real-time RT-PCR. Conclusion: We report for the first time that acute MI is associated with mobilization of non-hematopoietic VSELs expressing pluripotent stem cells markers.

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