scholarly journals Transcriptome-Wide Profile of 25-Hydroxyvitamin D3 in Primary Immune Cells from Human Peripheral Blood

Nutrients ◽  
2021 ◽  
Vol 13 (11) ◽  
pp. 4100
Author(s):  
Andrea Hanel ◽  
Igor Bendik ◽  
Carsten Carlberg
Keyword(s):  
Vitamin D ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Physiological Concentration ◽  
Peripheral Blood Mononuclear ◽  
Dihydroxyvitamin D3 ◽  
Interacting Protein

Vitamin D3 is an essential micronutrient mediating pleiotropic effects in multiple tissues and cell types via its metabolite 1α,25-dihydroxyvitamin D3 (1,25(OH)2D3), which activates the transcription factor vitamin D receptor. In this study, we used peripheral blood mononuclear cells (PBMCs) obtained from five healthy adults and investigated transcriptome-wide, whether the precursor of 1,25(OH)2D3, 25-hydroxyvitamin D3 (25(OH)D3), has gene regulatory potential on its own. Applying thresholds of >2 in fold change of gene expression and <0.05 as a false discovery rate, in this ex vivo approach the maximal physiological concentration of 25(OH)D3 (250 nM (nmol/L)) none of the study participants had a significant effect on their PBMC transcriptome. In contrast, 1000 and 10,000 nM 25(OH)D3 regulated 398 and 477 genes, respectively, which is comparable to the 625 genes responding to 10 nM 1,25(OH)2D3. The majority of these genes displayed specificity to the tested individuals, but not to the vitamin D metabolite. Interestingly, the genes MYLIP (myosin regulatory light chain interacting protein) and ABCG1 (ATP binding cassette subfamily G member 1) showed to be specific targets of 10,000 nM 25(OH)D3. In conclusion, 100- and 1000-fold higher 25(OH)D3 concentrations than the reference 10 nM 1,25(OH)2D3 are able to affect the transcriptome of PBMCs with a profile comparable to that of 1,25(OH)2D3.

scholarly journals MicroRNA levels quantified in whole blood varies from PBMCs

F1000Research ◽  
2014 ◽  
Vol 3 ◽  
pp. 183 ◽  
Author(s):  
Sadaf Atarod ◽  
Hannah Smith ◽  
Anne Dickinson ◽  
Xiao-Nong Wang
Keyword(s):  
Whole Blood ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Microrna Expression ◽  
Peripheral Blood Mononuclear ◽  
Total Rna ◽  
Disease Stages ◽  
Different Cell Types

MicroRNAs are non-coding RNAs that negatively regulate mRNA expression and play significant roles in both health and disease. Differential microRNA expression has been used to aid diagnosis and discriminate disease stages. The accuracy and reliability of microRNA expression measurement is of utmost importance. Quantification of microRNA expression in human peripheral blood is commonly detected using total RNA extracted via different methods. To date, no convincing data are available showing whether microRNA quantification results can be influenced by the use of total RNA extracted from whole blood or peripheral blood mononuclear cells (PBMCs). This study examined miR-146a-5p and miR-155-5p expression using total RNA extracted in parallel from whole blood and PBMCs of 14 healthy volunteers. The data showed that the quantification of miRNA using total RNA extracted from whole blood varied from that of PBMCs, indicating that the miRNA expression was a result of all the different cell-types present in whole blood. Our results suggested that the source of total RNA and the statistical analyses performed are crucial considerations when designing miRNA research.

scholarly journals MicroRNA levels quantified in whole blood varies from PBMCs

F1000Research ◽  
2015 ◽  
Vol 3 ◽  
pp. 183 ◽  
Author(s):  
Sadaf Atarod ◽  
Hannah Smith ◽  
Anne Dickinson ◽  
Xiao-Nong Wang
Keyword(s):  
Whole Blood ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Cell Types ◽  
Microrna Expression ◽  
Peripheral Blood Mononuclear ◽  
Total Rna ◽  
Disease Stages ◽  
Different Cell Types

MicroRNAs are non-coding RNAs that negatively regulate mRNA expression and play significant roles in both health and disease. Differential microRNA expression has been used to aid diagnosis and discriminate disease stages. The accuracy and reliability of microRNA expression measurement is of utmost importance. Quantification of microRNA expression in human peripheral blood is commonly detected using total RNA extracted via different methods. To date, no convincing data are available showing whether microRNA quantification results can be influenced by the use of total RNA extracted from whole blood or peripheral blood mononuclear cells (PBMCs). This study examined miR-146a-5p and miR-155-5p expression using total RNA extracted in parallel from whole blood and PBMCs of 14 healthy volunteers. The data showed that the quantification of miRNA using total RNA extracted from whole blood varied from that of PBMCs, indicating that the miRNA expression was a result of all the different cell-types present in whole blood. Our results suggested that the source of total RNA and the statistical analyses performed are crucial considerations when designing miRNA research.

A pathway analysis of poly(I:C)-induced global gene expression change in human peripheral blood mononuclear cells

2006 ◽  
Vol 26 (2) ◽  
pp. 125-133 ◽  
Author(s):  
C. Chris Huang ◽  
Karen E. Duffy ◽  
Lani R. San Mateo ◽  
Bernard Y. Amegadzie ◽  
Robert T. Sarisky ◽  
...  
Keyword(s):  
Gene Expression ◽  
Viral Infection ◽  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Poly I:C ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells

To gain global pathway perspective of ex vivo viral infection models using human peripheral blood mononuclear cells (PBMCs), we conducted expression analysis on PBMCs of healthy donors. RNA samples were collected at 3 and 24 h after PBMCs were challenged with the Toll-like receptor-3 (TLR3) agonist polyinosinic acid-polycytidylic acid [poly(I:C)] and analyzed by internally developed cDNA microarrays and TaqMan PCR. Our results demonstrate that poly(I:C) challenge can elicit certain gene expression changes, similar to acute viral infection. Hierarchical clustering revealed distinct immediate early, early-to-late, and late gene regulation patterns. The early responses were innate immune responses that involve TLR3, the NF-κB-dependent pathway, and the IFN-stimulated pathway, whereas the late responses were mostly cell-mediated immune response that involve activation of cell adhesion, cell mobility, and phagocytosis. Overall, our results expanded the utilities of this ex vivo model, which could be used to screen molecules that can modulate viral stress-induced inflammation, in particular those mediated via TLRs.

In vitro and ex vivo effect of tiaprofenic acid on human peripheral blood mononuclear cells

1992 ◽  
Vol 14 (7) ◽  
pp. 1279-1284
Author(s):  
Wilma Barcellini ◽  
Maria Orietta Borghi ◽  
Cristina Fain ◽  
Nicoletta Del Papa ◽  
Patrizia Favini ◽  
...  
Keyword(s):  
Peripheral Blood Mononuclear Cells ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Ex Vivo ◽  
Human Peripheral Blood ◽  
Tiaprofenic Acid ◽  
Peripheral Blood Mononuclear ◽  
Blood Mononuclear Cells
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