scholarly journals Sildenafil does not affect the proliferation of human lymphocytes in the in vitro transplant model

2019 ◽  
Author(s):  
Beata Kaleta
Keyword(s):  
Bone Marrow ◽  
Immune System ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Venous Blood ◽  
Human Immune System ◽  
Pde5 Inhibitors ◽  
Trypan Blue Exclusion ◽  
Peripheral Blood Mononuclear

Sildenafil is a selective inhibitor of type 5 phosphodiesterase (PDE5) used in the treatment of erectile dysfunction (ED) and pulmonary arterial hypertension (PAH). Numerous studies revealed beneficial effects of sildenafil use in chronic kidney disease, and also in renal, liver, heart and bone marrow transplant recipients. Some reports suggest that sildenafil modulates function of the immune system, and additionally proliferation of endothelial, bone marrow and cancer cells. Despite the fact that PDE5 inhibitors showed efficacy and safety, it is very important to know whether these drugs have immunomodulatory properties, because are used in patients after organ transplantation. The influence of sildenafil on antigen-induced proliferation of lymphocytes remains currently unknown, thus the aim of the study was to investigate the effects of the drug on human peripheral blood mononuclear cells (PBMCs) proliferation in a mixed lymphocyte reaction (MLR).PBMCs were isolated from venous blood from 30 donors. The proliferation was examined on the DNA synthesis level by measurements of 3H-thymidine incorporation. Cell viability was determined using trypan blue exclusion method.The study demonstated that sildenafil at concentrations of 0.06 µM, 0.6 µM and 6µM did not affect auto- and alloantigen-induced  proliferation of PBMCs and showed no cytotoxic effect. However, further analysis is required to fully understand the role of PDE5 inhibitors in the regulation of human immune system.

Immune Modulation by cross-linked Bovine Colostrum in vitro and in vivo

Pteridines ◽  
2010 ◽  
Vol 21 (1) ◽  
pp. 94-102
Author(s):  
Ninfa R. Pedersen ◽  
Budi J. Hidayat
Keyword(s):  
Atopic Dermatitis ◽  
Immune System ◽  
Immune Modulation ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Bovine Colostrum ◽  
Human Immune System ◽  
Mature Milk ◽  
Peripheral Blood Mononuclear ◽  
Tryptophan Degradation

Abstract Bovine colostrum (BC) is the early thick yellow fluid produced by cows during the first several days after birth of the calf. BC is different from "mature" milk as it contains nutrients and immune factors, which strengthen the immune system of the newborn calf in its first week of life. BC has a long history of use in traditional medicine throughout the world and is currently also used in a topical cream for the treatment of immuno-related skin problems such as atopic dermatitis or psoriasis. However, despite a large amount of literature concerning the properties of human or bovine colostrum, there are only few reports on the effects of colostrum on the human immune system. In this study, the effects of cross-linked BC containing hyaluronic acid on the T-cell/macrophage interplay were investigated in human peripheral blood mononuclear cells (PBMC) and the results were compared with the effects of euxyl 9010, which was used to preserve the BC ingredients. The cross-linked BC preparations showed significant immunomodulatory effects on unstimulated and phytohaemagglutinin (PHA)-stimulated human PBMC by modulating tryptophan degradation and formation of neopterin in a biphasic manner. These results could be relevant for some of the beneficial effects of BC observed in the treatment of skin lesions of patients with atopic dermatitis or psoriasis. However, additional studies are needed to confirm the first positive results obtained in patients.

scholarly journals The Comparison of the Immunologic Properties of Stem Cells Isolated from Human Exfoliated Deciduous Teeth, Dental Pulp, and Dental Follicles

2016 ◽  
Vol 2016 ◽  
pp. 1-15 ◽  
Author(s):  
Selin Yildirim ◽  
Noushin Zibandeh ◽  
Deniz Genc ◽  
Elif Merve Ozcan ◽  
Kamil Goker ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Dental Pulp ◽  
Fas Ligand ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Venous Blood ◽  
Deciduous Teeth ◽  
Peripheral Blood Mononuclear ◽  
Human Exfoliated Deciduous Teeth

Aim. To compare the effects of various mesenchymal stem cells, those isolated from human exfoliated deciduous teeth (SHEDs), dental pulp stem cells (DPSCs), and dental follicle stem cells (DFSCs), on human peripheral blood mononuclear cells (PBMCs).Method. Mesenchymal stem cells were isolated from three sources in the orofacial region. Characterization and PCR analyses were performed. Lymphocytes were isolated from healthy peripheral venous blood. Lymphocytes were cocultured with stem cells in the presence and absence of IFN-γand stimulated with anti-CD2, anti-CD3, and anti-CD28 for 3 days. Then, lymphocyte proliferation, the number of CD4+FoxP3+T regulatory cells, and the levels of Fas/Fas ligand, IL-4, IL-10, and IFN-γin the culture supernatant were measured.Results. The DFSCs exhibited an enhanced differentiation capacity and an increased number of CD4+FoxP3+T lymphocytes and suppressed the proliferation and apoptosis of PBMCs compared with SHEDs and DPSCs. The addition of IFN-γaugmented the proliferation of DFSCs. Furthermore, the DFSCs suppressed IL-4 and IFN-γcytokine levels and enhanced IL-10 levels compared with the other cell sources.Conclusion. These results suggest that IFN-γstimulates DFSCs by inducing an immunomodulatory effect on the PBMCs of healthy donors while suppressing apoptosis and proliferation and increasing the number of CD4+FoxP3+cells.

419. Rational design of bifunctional siRNAs that silence genes and recruit the innate immune system to treat ectopic pregnancies

2008 ◽  
Vol 20 (9) ◽  
pp. 99
Author(s):  
S. Tong ◽  
M. P. Gantier ◽  
M. Belhke ◽  
B. R. G. Williams
Keyword(s):  
Immune System ◽  
Gene Silencing ◽  
Rational Design ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Micro Rna ◽  
Peripheral Blood Mononuclear ◽  
Sirna Sequence ◽  
Sense Strand ◽  
Ectopic Pregnancies

RNA interference (RNAi) is a new therapeutic approach, silencing genes to disrupt diseases. However, short interfering siRNAs (molecule used in RNAi) can have off-target effects, activating the immune system through RNA sensing toll-like receptors (TLR) 3, 7 and 8. We have previously proposed that in some diseases (cancers, ectopic pregnancies) it may be useful to enhance the immune response. A novel class of immunostimulatory siRNAs could be developed, silencing genes important to disease and recruiting the immune system to further aid disease clearance. We set out to develop a rational design strategy that enhances immunostimulatory properties to any siRNA sequence but maintains effective gene silencing. We screened a set of siRNAs targeting lamin. All were of the same sequence, except for different immunostimulatory motifs on the 3′ end of the sense strand. We also investigated a different design where we added a small micro-RNA like poly-uridine bulge (potentially immunostimulatory) on the sense strand. We used human peripheral blood mononuclear cells (PBMCs) to test for immunostimulation, and HEK 293-T-cells to test for lamin gene knockdown.Of all strategies tested, the poly-uridine bulge was best. It silenced the lamin gene as effectively as control, but caused a 2–3 fold increase of IFN-α and TNF-α. We verified this approach by adding the poly-uridine bulge onto an siRNA of low immunostimulatory potential targeting GFP. The bulge markedly enhanced immunostimulation in a dose response manner, and did not compromise gene knockdown. The addition of a poly-uridine bulge to siRNAs can increase immunostimulation without affecting gene silencing efficacy. Immunostimulatory siRNAs might be particularly efficacious to treat ectopic pregnancies where there are abundant immune cells, and functional TLR 7/8 in the trophoblast (unpublished observations). We now plan to test this immunostimulatory siRNA approach in an in vivo ectopic pregnancy model using JEG-3 cells xenographted in NOD-SCID mice.

scholarly journals Control of hsp70 RNA levels in human lymphocytes.

1987 ◽  
Vol 104 (2) ◽  
pp. 183-187 ◽  
Author(s):  
L Kaczmarek ◽  
B Calabretta ◽  
H T Kao ◽  
N Heintz ◽  
J Nevins ◽  
...  
Keyword(s):  
Culture Medium ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Peripheral Blood Mononuclear ◽  
Growth State ◽  
Steady State Levels ◽  
Rna Blots

The expression of a hsp70 gene in human cells has previously been shown to be related to the growth state of the cells. As an alternative to in vitro synchronization procedures, we have measured steady-state levels of the RNA for a heat-shock protein 70 (hsp70) in human peripheral blood mononuclear cells (PBMC) that are naturally quiescent in a G0 state. The probe used recognized, on RNA blots, one single band. The levels of this hsp70 RNA are elevated in circulating PBMC and decrease when the cells are incubated with serum, or phytohemagglutinin, or simply when they are incubated in culture medium. The levels of hsp70 RNA decrease within 30 min after in vitro culture, and are accompanied by an increase in the levels of c-fos RNA. These findings, together with other recent reports in the literature, suggest a possible role of the hsp70 proteins in the regulation of cell growth.

Interaction of hypothalamic Na,K-ATPase inhibitor with isolated human peripheral blood mononuclear cells

1994 ◽  
Vol 14 (5) ◽  
pp. 231-242 ◽  
Author(s):  
Beatrice M. Anner ◽  
Danielle Lacotte ◽  
Rolf M. Anner ◽  
Marlis Moosmayer
Keyword(s):  
Cell Viability ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
Exchange Process ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells ◽  
Isolated Human Lymphocytes

A ligand for the digitalis receptor located on the membrane-embedded Na,K-ATPase (NKA; EC 3.6.1.37) has been isolated from bovine hypothalamus (hypothalamic inhibitory factor; HIF) and identified as isomeric ouabain (Tymiaket al, 1993,Proc. Natl. Acad. Sci.90: 8189–8193). In analogy to cardioactive steroids (CS) derived from plants or from toad, HIF inhibits the Na/K-exchange process and the ATPase activity of isolated Na,K-ATPase although by a different molecular action mechanism. In the present work we show that, as plant-derived ouabain, HIF inhibits86Rb-uptake by isolated human lymphocytes with an IC50 of about 20 nM; above this concentration HIF reduces cell viability in contrast to ouabain. The decrease in cell viability by excess HIF is accompanied by discrete morphological alterations (mitochondrial swelling) visible by transmission electron microscopy of ultra-thin sectioned peripheral blood mononuclear cells. Taken together the results show that the hypothalamic NKA inhibitor blocks NKA of isolated human lymphocytes with high potency at nanomolar concentrations without toxicity; concentrations exceeding the ones required to block86Rb-uptake reduce cell viability, probably due to leak formation across the NKA molecule. Thus, lymphocytes constitute a potential target for HIF action and by their altered NKA status a possible messenger between the nervous and the immune system.

scholarly journals Sexual-dimorphism in human immune system aging

2019 ◽  
Author(s):  
Eladio J. Márquez ◽  
Cheng-han Chung ◽  
Radu Marches ◽  
Robert J. Rossi ◽  
Djamel Nehar-Belaid ◽  
...  
Keyword(s):  
Immune System ◽  
Mononuclear Cells ◽  
Immune Cell ◽  
Rna Seq ◽  
Human Immune System ◽  
Peripheral Blood Mononuclear ◽  
Cell Functions ◽  
Age Related ◽  
Male Specific

AbstractDifferences in immune function and responses contribute to health- and life-span disparities between sexes. However, the role of sex in immune system aging is not well understood. Here, we characterize peripheral blood mononuclear cells from 172 healthy adults 22-93 years of age using ATAC-seq, RNA-seq, and flow-cytometry. These data reveal a shared epigenomic signature of aging including declining naïve T cell and increasing monocyte/cytotoxic cell functions. These changes were greater in magnitude in men and accompanied by a male-specific genomic decline in B-cell specific loci. Age-related epigenomic changes first spike around late-thirties with similar timing and magnitude between sexes, whereas the second spike is earlier and stronger in men. Unexpectedly, genomic differences between sexes increase after age 65, with men having higher innate and pro-inflammatory activity and lower adaptive activity. Impact of age and sex on immune cell genomes can be visualized at https://immune-aging.jax.org to provide insights into future studies.

scholarly journals A Novel cDNA-uPA/SCID/Rag2-/-/Jak3-/- Mouse Model for Hepatitis Virus Infection and Reconstruction of Human Immune System

2021 ◽  
Author(s):  
Takuro Uchida ◽  
Yuji Teraoka ◽  
Michio Imamura ◽  
Hatsue Fujino ◽  
Atsushi Ono ◽  
...  
Keyword(s):  
Immune System ◽  
Mouse Model ◽  
Mononuclear Cells ◽  
Hepatitis Virus ◽  
Human Leukocyte ◽  
Human Hepatocytes ◽  
Human Immune System ◽  
Peripheral Blood Mononuclear ◽  
Type Plasminogen Activator

Abstract The humanized mouse is a widely used in vivo model for the investigation of pathogenesis and drug development. In this study, we generated new immunodeficiency cDNA-urokinase-type plasminogen activator (uPA)/SCID/Rag2-/-/Jak3-/- mice and established the mouse model with a humanized liver and immune system. Transplantation of human hepatocytes with human leukocyte antigen (HLA)-A24 resulted in establishment of a highly replaced liver in this mouse model. These mice were successfully infected with hepatitis B virus and hepatis C virus (HCV) for a prolonged period and are available for the analysis of the effect of anti-HCV drugs. Administration of peripheral blood mononuclear cells (PBMCs) obtained from a donor with HLA-A24 resulted in the establishment of 22.6–81.3% of human CD45-positive mononuclear cell chimerism in liver infiltrating cells without causing graft versus host disease. When mice were transplanted with human hepatocytes and then administered PBMCs, an alloimmune response between transplanted human hepatocytes and PBMCs occurred, with production of transplanted hepatocyte-specific anti-HLA antibody. The alloimmunity was never inhibited by methylprednisolone nor cyclosporine A. In conclusion, we succeeded in establishing a humanized liver and immune system using a novel cDNA-uPA/SCID/Rag2-/-/Jak3-/- mouse. This model is not only useful to study hepatitis virus virology but also to study alloimmunity.

scholarly journals Assessment of genotoxic effects of a fungicide product and its active substances on human peripheral blood mononuclear cells

2019 ◽  
Vol 34 (1) ◽  
pp. 61-67
Author(s):  
Hayal Cobanoglu ◽  
Baver Coskun ◽  
Akin Cayir
Keyword(s):  
Dna Damage ◽  
Trypan Blue ◽  
Mononuclear Cells ◽  
Plant Protection ◽  
Human Peripheral Blood ◽  
Genotoxic Effects ◽  
Trypan Blue Exclusion ◽  
Peripheral Blood Mononuclear ◽  
Trypan Blue Exclusion Test ◽  
Active Substances

The genotoxic potential of the plant protection product Signum? and its two active substances (pyraclostrobin and boscalid) was investigated by the comet assay (pH>13). Leukocytes isolated from whole blood were treated with different concentrations (0.1-25 ?g/ml) of the fungicide for 2 h and 20 h. The Trypan Blue exclusion test showed higher cell viability (>84%) under all three concentrations and in the two treatment periods. The obtained results revealed that both Signum? and pyraclostrobin induced statistically significant DNA damage. In contrast, boscalid did not cause statistically significant DNA damage after 2 h exposure although it caused DNA damage at higher concentrations after a longer time exposure (20 h). It is deducible that pyraclostrobin and Signum? might be genotoxic. However, within the studied concentration ranges, none of the fungicides was found to be cytotoxic in the two treatment periods.

scholarly journals Identification of age and ethnicity specific gene expression biomarkers for immune aging

2021 ◽  
Author(s):  
Yang Hu ◽  
Yudai Xu ◽  
Lipeng Mao ◽  
Wen Lei ◽  
Jian Xiang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Immune System ◽  
Mononuclear Cells ◽  
Specific Gene ◽  
Human Immune System ◽  
Peripheral Blood Mononuclear ◽  
Network Analyses ◽  
Age Related ◽  
Human Immune ◽  
The Impact

ABSTRACTHuman immune system functions over an entire lifetime, yet how and why the immune system becomes less effective with age are not well understood. Here, we characterize peripheral blood mononuclear cells transcriptome from 172 healthy adults with 21~90 years of age using RNA-seq and the weighted gene correlation network analyses (WGCNA). These data reveal a set of insightful gene expression modules and representative gene biomarkers for human immune system aging from Asian and Caucasian ancestry, respectively. Among them, the aging-specific modules show an age-related gene expression variation spike around early-seventies. In addition, it is not known whether Asian and Caucasian immune systems go through similar gene expression changes throughout their lifespan, and to what extent these aging-associated changes are shared among ethnicities. We find the top hub genes including NUDT7, CLPB, OXNAD1 and MLLT3 are shared between Asian and Caucasian aging related modules and further validated in human PBMCs from different age groups. Overall, the impact of age and race on transcriptional variation elucidated from this study provide insights into the transcriptional driver of immune aging.

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