scholarly journals Beta-2 Microglobulin And Ubiquitin C Identified as Two Robust Housekeeping Genes for RNA Expression Normalization in Real Time PCR on Human Leukocytes and Regulatory T Cells

2020 ◽  
Vol 31 (4) ◽  
Author(s):  
Nadira Delhem
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Real Time ◽  
Real Time Pcr ◽  
Housekeeping Genes ◽  
Rna Expression ◽  
Human Leukocytes ◽  
Beta 2 ◽  
Beta 2 Microglobulin

scholarly journals Beta-2 microglobulin a robust reference housekeeping genes for RNA expression normalization in real time PCR on human leukocytes

2019 ◽  
Author(s):  
Clara MILHEM ◽  
Céline INGELAERE ◽  
Olivier MORALES ◽  
Nadira DELHEM
Keyword(s):  
Gene Expression ◽  
Real Time ◽  
Real Time Pcr ◽  
Housekeeping Genes ◽  
Internal Controls ◽  
Rna Expression ◽  
Rt Pcr ◽  
Genomic Study ◽  
Beta 2 ◽  
Beta 2 Microglobulin

Abstract Background: Changes in gene expression are increasingly used to evaluate the effects of exposure to environmental agents. Housekeeping genes (Hk) are essential in these analyzes as internal controls to normalize expression levels assessed in real-time PCR (RT-PCR). Ideal Hk genes (i) are constitutively expressed; (ii) do not respond to external stimuli and (iii) show small or no variation between samples or from one assay to another. Previous studies indicate that some commonly used Hk genes, including glyceraldehyde 3-phosphate dehydrogenase (GAPDH) and beta-actin, have differential expression in various cell lines. The objective of this study is to identify and validate the most appropriate housekeeping genes for RNA expression analysis of human primary peripheral blood mononuclear cells (PBMCs) in response to antigen like stimulation. Methods: The expression of the five most commonly used genes for a genomic study of lymphocytes was determined [HPRT, 18S, GAPDH, Ubiquitin C (UBC), beta-2 microglobulin (B2M)] on PBMCs, isolated from fresh blood, following activation with anti-CD3 and anti-CD28. This leads to the activation and proliferation of T lymphocytes in order to modulate the expression of specific genes. Results : Using reverse transcriptase RT-PCR protocol, we show that following activation only B2M remain unchanged in PBMCs. This result has been confirmes In contrast, 18S, HPRT and GAPDH show significant variation in PBMC gene expression. Conclusion: Although our results suggest that the relevance of Hk genes should be determined for each experimental condition, B2M appear to be excellent candidate as internal controls. Keywords : Housekeeping genes, RT-PCR, Human primary PBMC.

Analytical evaluation of a real-time PCR-based DNA demethylation assay to assess the frequency of naturally occurring regulatory T cells in peripheral blood

2016 ◽  
Vol 49 (15) ◽  
pp. 1173-1180 ◽  
Author(s):  
Maria Metzker ◽  
Maria Shipkova ◽  
Nicolas von Ahsen ◽  
Rainer Andag ◽  
Manabu Abe ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Real Time ◽  
Real Time Pcr ◽  
Peripheral Blood ◽  
Dna Demethylation ◽  
Analytical Evaluation ◽  
Naturally Occurring

scholarly journals Validation of housekeeping genes for normalizing RNA expression in real-time PCR

BioTechniques ◽  
2004 ◽  
Vol 37 (1) ◽  
pp. 112-119 ◽  
Author(s):  
Keertan Dheda ◽  
Jim F. Huggett ◽  
Stephen A. Bustin ◽  
Margaret A. Johnson ◽  
Graham Rook ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Housekeeping Genes ◽  
Rna Expression

scholarly journals Quantification of Regulatory T Cells in Septic Patients by Real-Time PCR–Based Methylation Assay and Flow Cytometry

PLoS ONE ◽  
2012 ◽  
Vol 7 (11) ◽  
pp. e49962 ◽  
Author(s):  
Roman Tatura ◽  
Michael Zeschnigk ◽  
Michael Adamzik ◽  
Michael Probst-Kepper ◽  
Jan Buer ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
T Cells ◽  
Regulatory T Cells ◽  
Real Time ◽  
Real Time Pcr ◽  
Methylation Assay

scholarly journals Identification and Validation of Suitable Housekeeping Genes for Normalizing Quantitative Real-Time PCR Assays in Injured Peripheral Nerves

PLoS ONE ◽  
2014 ◽  
Vol 9 (8) ◽  
pp. e105601 ◽  
Author(s):  
Giovanna Gambarotta ◽  
Giulia Ronchi ◽  
Olivier Friard ◽  
Pantaleo Galletta ◽  
Isabelle Perroteau ◽  
...  
Keyword(s):  
Real Time ◽  
Real Time Pcr ◽  
Peripheral Nerves ◽  
Housekeeping Genes ◽  
Quantitative Real Time Pcr ◽  
Pcr Assays

LCMV-specific, class II-restricted cytotoxic T cells in beta 2-microglobulin-deficient mice

Science ◽  
1992 ◽  
Vol 255 (5051) ◽  
pp. 1576-1578 ◽  
Author(s):  
D Muller ◽  
B. Koller ◽  
J. Whitton ◽  
K. LaPan ◽  
K. Brigman ◽  
...  
Keyword(s):  
T Cells ◽  
Cytotoxic T Cells ◽  
Class Ii ◽  
Specific Class ◽  
Beta 2 ◽  
Beta 2 Microglobulin ◽  
Deficient Mice
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