scholarly journals Profil Peripheral Blood Mononuclear Cells (PBMC) Pasien dengan Berbagai Usia Menggunakan Flow Cytometry di Klinik Hayandra

2018 ◽  
Vol 4 (4) ◽  
pp. 208
Author(s):  
Masayu Nadhira ◽  
Riris Lindiawati Puspitasari ◽  
Karina F Moegni ◽  
Imam Rosadi ◽  
Iis Rosliana
Keyword(s):  
Flow Cytometry ◽  
Natural Killer ◽  
Peripheral Blood Mononuclear Cells ◽  
Immune Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Group 2 ◽  
Cluster Of Differentiation

<p><em>Abstrak</em> - <strong>Profil Peripheral Blood Mononuclear Cells (PBMC) setiap orang dapat berbeda-beda. Salah satu faktor yang diduga mempengaruhi adanya perbedaan tersebut adalah usia. Manusia berusia lanjut lebih mudah terserang penyakit karena terjadinya penurunan fungsi dan jumlah sel imun. Oleh karena itu, perlu dilakukan analisis komposisi PBMC untuk mengetahui profil PBMC seseorang berdasarkan usia, sehingga dapat diketahui jumlah sel imun dalam tubuh manusia. Tujuan dari penelitian ini adalah untuk menganalisis profil PBMC pasien dengan berbagai usia di Klinik Hayandra menggunakan flow cytometry. Manfaat dilakukannya kegiatan penelitian ini adalah diperolehnya informasi mengenai profil PBMC pasien di Klinik Hayandra dan kaitannya dengan usia. Metode yang dilakukan diantaranya adalah isolasi PBMC, proses pewarnaan PBMC, dan penentuan populasi sel menggunakan flow cytometry. Cluster of Differentiation (CD) yang digunakan pada kegiatan penelitian ini diantaranya adalah CD3, CD19, CD8, CD56, dan CD314 atau disebut juga NKG2D (Natural Killer Group 2 member D). Hasil yang diperoleh pada kegiatan penelitian ini adalah ditemukannya korelasi negatif yang bermakna antara persentase jumlah sel limfosit T sitotoksik (CD3+/CD8+) dengan usia dan ditemukannya korelasi positif yang bermakna antara persentase jumlah sel NKT (CD3+/CD56+) dengan usia. Dengan demikian usia pasien dapat mempengaruhi jumlah sel imun pasien.</strong></p><p><em>Abstract</em> - <strong>Peripheral Blood Mononuclear Cells (PBMC) profiles of each person can different. One of the factors that allegedly affects the difference is age. Elderly human are more susceptible attacked disease due to decreased function and number of immune cells. Therefore, it is necessary to do analyze PBMC composition to know the PBMC profile of person, so that can be known the number of immune cells in the human body. The purpose of this research is to analyze the PBMC profile of patients with various ages in the Hayandra Clinic using flow cytometry. The benefit of doing this practical work is obtained information about the PBMC profile of patients in the Hayandra Clinic and its relation to age. Methods which are conducted among others are PBMC isolation, PBMC staining process, running flow cytometry, and determination of cell population. Cluster of Differentiation (CD) used in this practical work among others are CD3, CD19, CD8, CD56, and CD314 or also called NKG2D (Natural Killer Group 2 member D). The results obtained in this research were the found of significant negative correlation between percentage of cytotoxic T cell lymphocytes numbers (CD3+/CD8+) with age and found a significant positive correlation between percentage of NKT cell numbers (CD3+/CD56+) with age. So it can be concluded that the patient's age can affect the number of the patient immune cells.</strong></p><p><em><strong>Keywords</strong></em> - <em>Peripheral Blood Mononuclear Cells (PBMC), flow cytometry, age, Cluster of Differentiation (CD)</em></p>

scholarly journals Function of ex vivo stimulated Natural killer T cells from patients with chronic HIV-1 infection

2019 ◽  
Author(s):  
Prakash Thapa ◽  
Pramod Nehete ◽  
Hong He ◽  
Bharti Nehete ◽  
Stephanie J. Buchl ◽  
...  
Keyword(s):  
Natural Killer ◽  
Peripheral Blood Mononuclear Cells ◽  
Immune Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Nkt Cells ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Hiv 1

AbstractNatural killer T (NKT) cells are innate immune cells that are responsible for the first line of antiviral defense, through crosstalk with downstream antigen-presenting cells, natural killer cells, and adaptive immune cells. Previous studies have indicated that NKT cell function is severely impaired in patients with chronic HIV-1 infection. It was reported that alpha-galactosylceramide, a potent agonist antigen for NKT cells, failed to trigger the expansion of NKT cells, or the production of anti-viral cytokines by NKT cells from HIV-1 infected patients in an in vitro assay, in which peripheral blood mononuclear cells (PBMCs) were cultured in the presence of alpha-galactosylceramide. In this study, we stimulated banked peripheral blood mononuclear cells from HIV-1-infected patients with dendritic cells (DC) generated ex vivo and loaded with alpha-galactosylceramide. The results showed that NKT cells were expanded in HIV infected subjects except in patients with advanced AIDS. Expanded NKT cells were capable of producing antiviral cytokines. Our results indicate that NKT cells in HIV infected individuals are potential targets for therapeutic intervention.

Efficacy of intrauterine administration of autologous peripheral blood mononuclear cells prior to embryo transfer in patients with recurrent implantation failures in assisted reproductive technology programmes

GYNECOLOGY ◽  
2018 ◽  
Vol 20 (2) ◽  
pp. 28-33
Author(s):  
T S Amyan ◽  
S G Perminova ◽  
L V Krechetova ◽  
V V Vtorushina
Keyword(s):  
Embryo Transfer ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Study Objective ◽  
Study Results ◽  
Blood Mononuclear Cells ◽  
History Of ◽  
Group 2

Study objective. To evaluate the efficacy of intrauterine administration of autologous peripheral blood mononuclear cells (PBMC) prior to embryo transfer in patients with recurrent implantation failures in IVF program. Materials and methods. The study enrolled 129 patients with recurrent implantation failures in an IVF programme. Group 1 - 42 patients who had intrauterine administration of autologous PBMC activated with hCG (Pregnyl 500 IU). Group 2 - 42 patients who had intrauterine administration of autologous PBMC without hCG activation. Group 3 (placebo) - 45 patients who had intrauterine administration of saline. Study results. In the hCG-activated PBMC group, the rates of positive blood hCG tests, implantation, and clinical pregnancy were significantly higher than the respective rates in the non-activated PBMC group and in the placebo group, both in a stimulated cycle and in an FET cycle (р≤0.05). Conclusion. Intrauterine administration of autologous PBMC prior to embryo transfer in an IVF/ICSI programme increases the efficacy of IVF program in patients with a history of recurrent implantation failures.

scholarly journals Comprehensive Evaluation of the Effects of Long-term Cryopreservation on Peripheral Blood Mononuclear Cells using Flow Cytometry

2021 ◽  
Author(s):  
Bo Li ◽  
Chunmei Yang ◽  
Gui Ja ◽  
Yansheng Liu ◽  
Na Wang ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Peripheral Blood Mononuclear ◽  
Effective Utilization ◽  
Blood Mononuclear Cells ◽  
Important Evidence

Abstract Human peripheral blood mononuclear cells (PBMCs) originate from hematopoietic stem cells (HSCs) in the bone marrow, which mainly includes lymphocytes (T cells, B cells, and natural killer [NK] cells) and monocytes. Cryopreserved PBMCs providing biobank resources are crucial for clinical application or scientific research. Here, we used flow cytometry to explore the influence of long-term cryopreservation on the quality of PBMCs with the aim of providing important evidence for the effective utilization of biobank resources. The PBMCs were isolated from the peripheral blood, which was collected from volunteers in the hospital. After long-term cryopreservation in liquid nitrogen, we analyzed the changes in cell numbers, viability, and multiple subtypes of PBMCs and studied the apoptosis, proliferation, activation, function, and status of T cells in comparison with freshly isolated PBMCs by flow cytometry, and then further tracked the effects of long-term cryopreservation on the same sample. Although the different cell types in the PBMCs dynamically changed compared with those in the freshly isolated samples, PBMC recovery and viability remained stable after long-term cryopreservation, and the number of most innate immune cells (e.g., monocytes and B cells) was significantly reduced compared to that of the freshly isolated PBMCs or long-term cryopreserved PBMCs; more importantly, the proportion of T cell subtypes, apoptosis, proliferation, and functional T cells, except for Tregs, were not affected by long-term cryopreservation. However, the proportions of activated T, naïve T, central memory T, effector T, and effector memory T cells dynamically changed after long-term cryopreservation. This article provides important evidence for the effective utilization of biobank resources. Long-term cryopreserved PBMCs can be partly used as biological resources for clinical research or basic studies, but the effect of cryopreservation on PBMCs should be considered when selecting cell samples, especially in research relating to activating or inhibiting function.

scholarly journals Statin-induced microRNAome alterations modulating inflammation pathways of peripheral blood mononuclear cells in patients with hypercholesterolemia

2020 ◽  
Vol 40 (9) ◽  
Author(s):  
Hung-Ju Lin ◽  
Sung-Liang Yu ◽  
Ta-Chen Su ◽  
Hsiu-Ching Hsu ◽  
Ming-Fong Chen ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Immune Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Transforming Growth Factor ◽  
Quantitative Polymerase Chain Reaction ◽  
Density Lipoprotein ◽  
Cardiovascular Risks ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Abstract Statins inhibit cholesterol biogenesis and modulate atheroma inflammation to reduce cardiovascular risks. Promoted by immune and non-immune cells, serum C-reactive protein (CRP) might be a biomarker suboptimal to assess inflammation status. Although it has been reported that statins modulated inflammation via microRNAs (miRNAs), evidence remains lacking on comprehensive profiling of statin-induced miRNAome alterations in immune cells. We recruited 19 hypercholesterolemic patients receiving 2 mg/day pitavastatin and 15 ones receiving 10 mg/day atorvastatin treatment for 12 weeks, and performed microarray-based profiling of 1733 human mature miRNAs in peripheral blood mononuclear cells (PBMCs) before and after statin treatment. Differentially expressed miRNAs were determined if their fold changes were &gt;1.50 or &lt;0.67, after validated using quantitative polymerase chain reaction (qPCR). The miRSystem and miTALOS platforms were utilized for pathway analysis. Of the 34 patients aged 63.7 ± 6.2 years, 27 were male and 19 were with coronary artery disease. We discovered that statins induced differential expressions of miR-483-5p, miR-4667-5p, miR-1244, and miR-3609, with qPCR-validated fold changes of 1.74 (95% confidence interval, 1.33–2.15), 1.61 (1.25–1.98), 1.61 (1.01–2.21), and 1.68 (1.19–2.17), respectively. The fold changes of the four miRNAs were not correlated with changes of low-density-lipoprotein cholesterol or CRP, after sex, age, and statin type were adjusted. We also revealed that RhoA and transforming growth factor-β signaling pathways might be regulated by the four miRNAs. Given our findings, miRNAs might be involved in statin-induced inflammation modulation in PBMCs, providing likelihood to assess and reduce inflammation in patients with atherosclerotic cardiovascular diseases.

Norepinephrine Inhibits Energy Metabolism of Human Peripheral Blood Mononuclear Cells via Adrenergic Receptors

2001 ◽  
Vol 21 (5) ◽  
pp. 627-635 ◽  
Author(s):  
Jan D. Lünemann ◽  
Frank Buttgereit ◽  
Robert Tripmacher ◽  
Christoph G. O. Baerwald ◽  
Gerd-Rüdiger Burmester ◽  
...  
Keyword(s):  
Energy Metabolism ◽  
Peripheral Blood Mononuclear Cells ◽  
Immune Cells ◽  
Peripheral Blood ◽  
Adrenergic Receptors ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Peripheral Blood Mononuclear ◽  
Immune Effector ◽  
Blood Mononuclear Cells

Previous studies demonstrated that the adaptive response to stressors and inflammatory signals involves the activation of the automotic nervous system. Catecholamines have been shown to modulate the activity of various immune effector cells directly via membrane adrenergic receptors. Here, we investigated immediate effects of norepinephrine on energy metabolism of immune cells. Norepinephrine inhibits oxygen consumption of human peripheral blood mononuclear cells at concentrations that are relevant to its physiological range. The ?-adrenoreceptor antagonist propranolol, but not the ?-adrenoreceptor antagonist phentolamine reversed the norepinephrine induced inhibition in quiescent cells. Conversely, phentolamine but not propranolol is capable of blocking norepinephrine mediated effects in mitogen activated human peripheral blood mononuclear cells. Our data indicate that the sensitization of ?- and ?-adrenoreceptors on immune cells is differentially regulated, and that these processes depend on the activation state of these cells. These findings have important implications for the understanding of stress-induced suppression of immune function and may contribute to the elucidation of the pathogenesis of immunologically mediated diseases.

scholarly journals Optimizing recovery of frozen human peripheral blood mononuclear cells for flow cytometry

PLoS ONE ◽  
2017 ◽  
Vol 12 (11) ◽  
pp. e0187440 ◽  
Author(s):  
Bo Langhoff Hønge ◽  
Mikkel Steen Petersen ◽  
Rikke Olesen ◽  
Bjarne Kuno Møller ◽  
Christian Erikstrup
Keyword(s):  
Flow Cytometry ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

Impaired natural killer cytotoxicity in peripheral blood mononuclear cells in Graves' disease

1995 ◽  
Vol 132 (2) ◽  
pp. 175-180 ◽  
Author(s):  
Mónica Marazuela ◽  
Juan A Vargas ◽  
Melchor Alvarez-Mon ◽  
Fernando Albarrán ◽  
Tomás Lucas ◽  
...  
Keyword(s):  
Natural Killer ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Natural Killer Activity ◽  
Graves Disease ◽  
Nk Activity ◽  
Peripheral Blood Mononuclear ◽  
Killer Activity ◽  
Blood Mononuclear Cells

Marazuela M, Vargas JA, Alvarez-Mon M, Albarrán F, Lucas T, Durántez A. Impaired natural killer cytotoxicity in peripheral blood mononuclear cells in Graves' disease. Eur J Endocrinol 1995;132:175–80. ISSN 0804–4643 We studied the natural killer (NK) activity of peripheral blood mononuclear cells (PBMC) in patients with Graves' disease (GD). Peripheral blood mononuclear cells from 20 untreated hyperthyroid patients with GD showed a significantly reduced NK activity against 51 Cr-labeled K562 cells (33.9 ± 15.9%), while in 32 euthyroid patients under antithyroid drug therapy, NK activity was similar to that of controls (46.9 ± 17.3 and 49.9 ± 20.2%, respectively). Furthermore, normalization of thyroid function with antithyroid drugs was associated with a significant increase and normalization of NK activity during the follow-up of nine GD patients (from 29.2 ± 17.9 to 48.1 ± 16.5%). This phenomenon could not be ascribed to a defective number of NK cells because the amounts of CD56 + and CD16 + cells in PBMC from both hyperthyroid and euthyroid GD patients were within normal ranges. Natural killer activity of PBMC from patients with toxic multinodular goiter was similar to that of normal controls (45 ± 12.8 to 49.9 ± 20%). No correlation was found between natural killer activity and serum levels of free thyroxine, TSH-inhibitory immunoglobulins, thyroidal antibodies to thryoglobulin and thyroidal microsomal antigen, dose or duration of antithyroid drug therapy. Natural killer activity from both controls and GD patients was enhanced in vitro by addition of recombinant interleukin 2 (IL-2), reaching control levels in hyperthyroid patients. These abnormalities were not associated with a defective IL-2 production by T cells, nor with a decreased IL-2R expression. We conclude that in untreated Graves' disease there is a decrease in NK cell activity in PBMC, probably dependent on the autoimmune process. Possible biological and clinical implications are discussed. Monica Marazuela, Hospital de la Princesa, c/Diego de Léon 62, Madrid 28006, Spain

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