LIVIVO - Search results -

Search results

Result 1 - 10 of total 21

Search options

Article ; Online: Evaluation of Cell Proliferation and Apoptosis in Immunotoxicity Testing.

Nagarkatti, Mitzi / Rieder, Sadiye Amcaoglu / Nagarkatti, Prakash S

Methods in molecular biology (Clifton, N.J.)

2018 Volume 1803, Page(s) 209–230

Abstract: Immunotoxicity testing is important in determining the toxic effects of various chemicals on the immune system. The immune system is a direct target of numerous toxicants, and the adverse effects include serious health complications such as ... ...

Abstract	Immunotoxicity testing is important in determining the toxic effects of various chemicals on the immune system. The immune system is a direct target of numerous toxicants, and the adverse effects include serious health complications such as susceptibility to infections, cancer, allergic reactions, and autoimmune diseases. One way to investigate the harmful effects of different chemicals is to study apoptosis and/or proliferation in immune cells. Apoptosis is defined as programmed cell death, and in general, this process helps in development and maintenance of tolerance and homeostasis. However, in the case of an insult by a toxicant, enhanced apoptosis of immune cells may cause immunosuppression resulting in the development of cancer and the inability to fight infections. Apoptosis is characterized by cell shrinkage, nuclear condensation, changes in cell membrane and mitochondria, DNA fragmentation, and protein degradation by caspases. Various methods are employed to investigate apoptosis, including direct measurement of apoptotic cells with flow cytometry and in situ labeling, as well as RNA, DNA, and protein assays that are indicative of apoptotic molecules. In addition to apoptosis, quantification of cell proliferation can provide important additional information about the effect of a toxicant upon various immune cell populations. In some cases, a toxicant may act as a mitogen pushing the immune cell into the different stages of the cell cycle. There are four stages of the active cell cycle: G1, S, G2, and M, with cell division occurring in M stage. Proliferation can be quantified by numerous methods, including staining with ki-67 or CFSE, BrdU labeling, MTT assay, and/or ATP quantification.
MeSH term(s)	Adenosine Triphosphate/metabolism ; Animals ; Apoptosis ; Caspases/metabolism ; Cell Proliferation ; DNA Fragmentation ; Gene Expression Regulation ; Humans ; Immune System/pathology ; Ki-67 Antigen/metabolism ; Membrane Potential, Mitochondrial ; Toxicity Tests/methods
Chemical Substances	Ki-67 Antigen ; Adenosine Triphosphate (8L70Q75FXE) ; Caspases (EC 3.4.22.-)
Language	English
Publishing date	2018-06-07
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-8549-4_14
Database	MEDical Literature Analysis and Retrieval System OnLINE

Full text online

Accessible to users with ZB MED library card

Order via subito

This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

Inter-library loan at ZB MED

Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

Details ▾
- Full text online
- Order with fees

Article ; Online: Eos, goddess of treg cell reprogramming.

Rieder, Sadiye Amcaoglu / Shevach, Ethan M

Immunity

2013 Volume 38, Issue 5, Page(s) 849–850

MeSH term(s)	Animals ; Carrier Proteins/metabolism ; Ikaros Transcription Factor/metabolism ; Interleukin-6/metabolism ; Nerve Tissue Proteins/metabolism ; T-Lymphocytes, Helper-Inducer/immunology
Chemical Substances	Carrier Proteins ; Interleukin-6 ; Nerve Tissue Proteins ; Zfpn1a4 protein, mouse ; Ikaros Transcription Factor (148971-36-2)
Language	English
Publishing date	2013-05-23
Publishing country	United States
Document type	Comment ; Journal Article
ZDB-ID	1217235-2
ISSN	1097-4180 ; 1074-7613
ISSN (online)	1097-4180
ISSN	1074-7613
DOI	10.1016/j.immuni.2013.05.001
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Zs.A 4227: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 69: Show issues

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article ; Online: Targeting the CD40-CD40L pathway in autoimmune diseases: Humoral immunity and beyond.

Karnell, Jodi L / Rieder, Sadiye Amcaoglu / Ettinger, Rachel / Kolbeck, Roland

Advanced drug delivery reviews

2018 Volume 141, Page(s) 92–103

Abstract: CD40 is a TNF receptor superfamily member expressed on both immune and non-immune cells. Interactions between B cell-expressed CD40 and its binding partner, CD40L, predominantly expressed on activated CD4+ T cells, play a critical role in promoting ... ...

Abstract	CD40 is a TNF receptor superfamily member expressed on both immune and non-immune cells. Interactions between B cell-expressed CD40 and its binding partner, CD40L, predominantly expressed on activated CD4+ T cells, play a critical role in promoting germinal center formation and the production of class-switched antibodies. Non-hematopoietic cells expressing CD40 can also engage CD40L and trigger a pro-inflammatory response. This article will highlight what is known about the biology of the CD40-CD40L axis in humans and describe the potential contribution of CD40 signaling on both hematopoietic and non-hematopoietic cells to autoimmune disease pathogenesis. Additionally, novel therapeutic approaches to target this pathway, currently being evaluated in clinical trials, are discussed.
MeSH term(s)	Animals ; Autoimmune Diseases/drug therapy ; Autoimmune Diseases/immunology ; CD40 Antigens/immunology ; CD40 Ligand/immunology ; Humans ; Immunity, Cellular ; Immunity, Humoral ; Signal Transduction
Chemical Substances	CD40 Antigens ; CD40 Ligand (147205-72-9)
Language	English
Publishing date	2018-12-13
Publishing country	Netherlands
Document type	Journal Article ; Review
ZDB-ID	639113-8
ISSN	1872-8294 ; 0169-409X
ISSN (online)	1872-8294
ISSN	0169-409X
DOI	10.1016/j.addr.2018.12.005
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Zs.A 2241: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (1.OG)
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article: CD28 family of receptors inter-connect in the regulation of T-cells.

Krueger, Janna / Jules, Felix / Rieder, Sadiye Amcaoglu / Rudd, Christopher E

Receptors & clinical investigation

2017 Volume 4

Abstract: T-cell activation is mediated by a combination of signals from the antigen receptor (TCR) and co-receptors such as CD28, cytotoxic T-lymphocyte antigen-4 (CTLA-4), programmed cell death antigen 1 (PD-1), CD28H and others. Each is a member of the CD28 ... ...

Abstract	T-cell activation is mediated by a combination of signals from the antigen receptor (TCR) and co-receptors such as CD28, cytotoxic T-lymphocyte antigen-4 (CTLA-4), programmed cell death antigen 1 (PD-1), CD28H and others. Each is a member of the CD28 receptor gene family. CD28 sends positive signals that promote T-cell responses, while CTLA-4 and PD-1 limit responses. It is the balance between these positive and negative signals that determines the amplitude and level of T-cell responses. The regulatory role of other family members is also becoming the focus of increasing interest. The function of certain CD28 family members such as CTLA-4 and PD-1 is dependent the expression of CD28. Together, these findings have important implications in generation of immune responses and the application of anti-receptor blocking reagents in immunotherapy.
Language	English
Publishing date	2017-09-25
Publishing country	United States
Document type	Journal Article
ISSN	2330-0558
ISSN	2330-0558
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

Inter-library loan at ZB MED

Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

Article ; Online: B7-H7 (HHLA2) inhibits T-cell activation and proliferation in the presence of TCR and CD28 signaling.

Rieder, Sadiye Amcaoglu / Wang, Jingya / White, Natalie / Qadri, Ariful / Menard, Catherine / Stephens, Geoffrey / Karnell, Jodi L / Rudd, Christopher E / Kolbeck, Roland

Cellular & molecular immunology

2020 Volume 18, Issue 6, Page(s) 1503–1511

Abstract: Modulation of T-cell responses has played a key role in treating cancers and autoimmune diseases. Therefore, understanding how different receptors on T cells impact functional outcomes is crucial. The influence of B7-H7 (HHLA2) and CD28H (TMIGD2) on T- ... ...

Abstract	Modulation of T-cell responses has played a key role in treating cancers and autoimmune diseases. Therefore, understanding how different receptors on T cells impact functional outcomes is crucial. The influence of B7-H7 (HHLA2) and CD28H (TMIGD2) on T-cell activation remains controversial. Here we examined global transcriptomic changes in human T cells induced by B7-H7. Stimulation through TCR with OKT3 and B7-H7 resulted in modest fold changes in the expression of select genes; however, these fold changes were significantly lower than those induced by OKT3 and B7-1 stimulation. The transcriptional changes induced by OKT3 and B7-H7 were insufficient to provide functional stimulation as measured by evaluating T-cell proliferation and cytokine production. Interestingly, B7-H7 was coinhibitory when simultaneously combined with TCR and CD28 stimulation. This inhibitory activity was comparable to that observed with PD-L1. Finally, in physiological assays using T cells and APCs, blockade of B7-H7 enhanced T-cell activation and proliferation, demonstrating that this ligand acts as a break signal. Our work defines that the transcriptomic changes induced by B7-H7 are insufficient to support full costimulation with TCR signaling and, instead, B7-H7 inhibits T-cell activation and proliferation in the presence of TCR and CD28 signaling.
MeSH term(s)	CD28 Antigens/metabolism ; Cell Proliferation ; Gene Expression Regulation ; Humans ; Immunoglobulins/metabolism ; Lymphocyte Activation/immunology ; Lymphocyte Culture Test, Mixed ; Models, Biological ; Protein Binding ; Receptors, Antigen, T-Cell/metabolism ; Signal Transduction ; T-Lymphocytes/cytology ; T-Lymphocytes/immunology
Chemical Substances	CD28 Antigens ; HHLA2 protein, human ; Immunoglobulins ; Receptors, Antigen, T-Cell
Language	English
Publishing date	2020-01-31
Publishing country	China
Document type	Journal Article
ZDB-ID	2435097-7
ISSN	2042-0226 ; 1672-7681
ISSN (online)	2042-0226
ISSN	1672-7681
DOI	10.1038/s41423-020-0361-7
Database	MEDical Literature Analysis and Retrieval System OnLINE

Full text online

Accessible to users with ZB MED library card

In stock of ZB MED Cologne/Königswinter

Zs.A 6681: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾

Article ; Online: Endotoxin reduction in protein solutions using octyl β-D-1-thioglucopyranoside wash on chromatography media.

Gadre, Dhanesh / Carlson, Marcia / Mullan, Ashley / Kabundi, Ivy / Pedowitz, Nichole / Rieder, Sadiye Amcaoglu / White, Natalie / Yu, Kathy / O'Connor, Ellen

Journal of chromatography. A

2018 Volume 1575, Page(s) 49–58

Abstract: Endotoxins are complex molecules and one of the most challenging impurities requiring separation in biopharmaceutical protein purification processes. Usually these contaminants are cleared during the downstream process, but if endotoxin interacts with ... ...

Abstract	Endotoxins are complex molecules and one of the most challenging impurities requiring separation in biopharmaceutical protein purification processes. Usually these contaminants are cleared during the downstream process, but if endotoxin interacts with the target protein it becomes difficult to remove. In the present study we identified a detergent, octyl-β-D-1-thioglucopyranoside (OTG), that disrupted endotoxin-protein interactions. The integration of this detergent into washes on several chromatography media was demonstrated to provide a separation tool for decreasing endotoxin from target proteins. This study also examined the mechanism of OTG endotoxin-protein disruption through phase modification incubation and chromatographic studies. The non-ionic OTG wash was shown to break both hydrophobic and electrostatic interactions between the endotoxin and protein. This mechanism contrasts with the breaking of hydrophobic interactions by washing with known endotoxin decreasing Triton X-100 detergent. The difference in mechanisms likely results in the ability of OTG to decrease endotoxin to levels less than those resulting from a detergent wash such as Triton X-100. Finally, we show the impact of the OTG endotoxin removal tool on the biopharmaceutical industry. While maintaining monomer purity and activity levels, endotoxin removal from a fusion protein allowed for decreased background levels in a T cell functional assay. The lowered baseline of T cell responses allowed for more effective detection of molecular interaction with the cells. The detergent wash can be used to both decrease the overall level of endotoxin in a purified protein solution and to enable more effective screening of lead candidate molecules.
MeSH term(s)	Chemistry, Pharmaceutical/methods ; Chromatography, Affinity ; Endotoxins/chemistry ; Endotoxins/isolation & purification ; Octoxynol/chemistry ; Thioglucosides/chemistry
Chemical Substances	Endotoxins ; Thioglucosides ; Octoxynol (9002-93-1)
Language	English
Publishing date	2018-09-18
Publishing country	Netherlands
Document type	Journal Article
ZDB-ID	1171488-8
ISSN	1873-3778 ; 0021-9673
ISSN (online)	1873-3778
ISSN	0021-9673
DOI	10.1016/j.chroma.2018.09.021
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Zs.A 813: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (1.OG)
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article ; Online: Staphylococcal enterotoxin B-induced microRNA-155 targets SOCS1 to promote acute inflammatory lung injury.

Rao, Roshni / Rieder, Sadiye Amcaoglu / Nagarkatti, Prakash / Nagarkatti, Mitzi

Infection and immunity

2014 Volume 82, Issue 7, Page(s) 2971–2979

Abstract: Staphylococcal enterotoxin B (SEB) causes food poisoning in humans. It is considered a biological weapon, and inhalation can trigger lung injury and sometimes respiratory failure. Being a superantigen, SEB initiates an exaggerated inflammatory response. ... ...

Abstract	Staphylococcal enterotoxin B (SEB) causes food poisoning in humans. It is considered a biological weapon, and inhalation can trigger lung injury and sometimes respiratory failure. Being a superantigen, SEB initiates an exaggerated inflammatory response. While the role of microRNAs (miRNAs) in immune cell activation is getting increasing recognition, their role in the regulation of inflammatory disease induced by SEB has not been studied. In this investigation, we demonstrate that exposure to SEB by inhalation results in acute inflammatory lung injury accompanied by an altered miRNA expression profile in lung-infiltrating cells. Among the miRNAs that were significantly elevated, miR-155 was the most overexpressed. Interestingly, miR-155(-/-) mice were protected from SEB-mediated inflammation and lung injury. Further studies revealed a functional link between SEB-induced miR-155 and proinflammatory cytokine gamma interferon (IFN-γ). Through the use of bioinformatics tools, suppressor of cytokine signaling 1 (SOCS1), a negative regulator of IFN-γ, was identified as a potential target of miR-155. While miR-155(-/-) mice displayed increased expression of Socs1, the overexpression of miR-155 led to its suppression, thereby enhancing IFN-γ levels. Additionally, the inhibition of miR-155 resulted in restored Socs1expression. Together, our data demonstrate an important role for miR-155 in promoting SEB-mediated inflammation in the lungs through Socs1 suppression and suggest that miR-155 may be an important target in preventing SEB-mediated inflammation and tissue injury.
MeSH term(s)	Acute Lung Injury/chemically induced ; Acute Lung Injury/metabolism ; Animals ; Enterotoxins/pharmacology ; Female ; Gene Expression Regulation/drug effects ; Gene Expression Regulation/immunology ; Interferon-gamma/genetics ; Interferon-gamma/metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; MicroRNAs/genetics ; MicroRNAs/metabolism ; Signal Transduction ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling Proteins/genetics ; Suppressor of Cytokine Signaling Proteins/metabolism
Chemical Substances	Enterotoxins ; MicroRNAs ; Mirn155 microRNA, mouse ; Socs1 protein, mouse ; Suppressor of Cytokine Signaling 1 Protein ; Suppressor of Cytokine Signaling Proteins ; enterotoxin B, staphylococcal (39424-53-8) ; Interferon-gamma (82115-62-6)
Language	English
Publishing date	2014-04-28
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	218698-6
ISSN	1098-5522 ; 0019-9567
ISSN (online)	1098-5522
ISSN	0019-9567
DOI	10.1128/IAI.01666-14
Database	MEDical Literature Analysis and Retrieval System OnLINE

In stock of ZB MED Cologne/Königswinter

Zs.A 684: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
Jg. 1995 - 2021: Lesesall (1.OG)
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾
- See ZB MED holdings
- Order with fees

Article ; Online: Multiple anti-inflammatory pathways triggered by resveratrol lead to amelioration of staphylococcal enterotoxin B-induced lung injury.

Rieder, Sadiye Amcaoglu / Nagarkatti, Prakash / Nagarkatti, Mitzi

British journal of pharmacology

2012 Volume 167, Issue 6, Page(s) 1244–1258

Abstract: Background and purpose: Inhalation of the superantigen,staphylococcal enterotoxin B (SEB), leads to the activation of the host T and invariant natural killer (iNK) T cells, thereby resulting in acute lung inflammation and respiratory failure but the ... ...

Abstract	Background and purpose: Inhalation of the superantigen,staphylococcal enterotoxin B (SEB), leads to the activation of the host T and invariant natural killer (iNK) T cells, thereby resulting in acute lung inflammation and respiratory failure but the underlying mechanism(s) of disease remain elusive, with limited treatment options. In this study, we investigated the therapeutic effectiveness of resveratrol, a plant polyphenol, during SEB-induced lung inflammation. Experimental approach: C57BL/6 mice were exposed to SEB (50 µg·per mouse), administered intranasally, and were treated with resveratrol (100 mg·kg(-1)) before or after SEB exposure. Lung injury was studied by measuring vascular permeability, histopathological examination, nature of infiltrating cells, inflammatory cytokine induction in the bronchoalveolar fluid (BALF), apoptosis in SEB-activated T cells and regulation of SIRT1 and NF-κB signalling pathways. Key results: Pretreatment and post-treatment with resveratrol significantly reduced SEB-induced pulmonary vascular permeability, and inflammation. Resveratrol significantly reduced lung infiltrating cells and attenuated the cytokine storm in SEB-exposed mice, which correlated with increased caspase-8-dependent apoptosis in SEB-activated T cells. Resveratrol treatment also markedly up-regulated Cd11b+ and Gr1+ myeloid-derived suppressor cells (MDSCs) that inhibited SEB-mediated T cell activation in vitro. In addition, resveratrol treatment was accompanied by up-regulation of SIRT1 and down-regulation of NF-κB in the inflammatory cells of the lungs. Conclusions and implications: The current study demonstrates that resveratrol may constitute a novel therapeutic modality to prevent and treat SEB-induced lung inflammation inasmuch because it acts through several pathways to reduce pulmonary inflammation.
MeSH term(s)	Administration, Intranasal ; Animals ; Anti-Inflammatory Agents/therapeutic use ; Apoptosis ; Cytokines/immunology ; Endothelial Cells/drug effects ; Enterotoxins/administration & dosage ; Female ; Lung/cytology ; Lung/immunology ; Lung Injury/drug therapy ; Lung Injury/immunology ; Lung Injury/pathology ; Mice ; Mice, Inbred C57BL ; Pneumonia/drug therapy ; Pneumonia/immunology ; Pneumonia/pathology ; Resveratrol ; Sirtuin 1/immunology ; Spleen/cytology ; Stilbenes/therapeutic use ; T-Lymphocytes/drug effects ; T-Lymphocytes/immunology ; Transcription Factor RelA/immunology
Chemical Substances	Anti-Inflammatory Agents ; Cytokines ; Enterotoxins ; Stilbenes ; Transcription Factor RelA ; enterotoxin B, staphylococcal (39424-53-8) ; Sirt1 protein, mouse (EC 3.5.1.-) ; Sirtuin 1 (EC 3.5.1.-) ; Resveratrol (Q369O8926L)
Language	English
Publishing date	2012-05-30
Publishing country	England
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	80081-8
ISSN	1476-5381 ; 0007-1188
ISSN (online)	1476-5381
ISSN	0007-1188
DOI	10.1111/j.1476-5381.2012.02063.x
Database	MEDical Literature Analysis and Retrieval System OnLINE

Full text online

Accessible to users with ZB MED library card

In stock of ZB MED Cologne/Königswinter

Uf I Zs.146: Show issues

Location:
Je nach Verfügbarkeit (siehe Angabe bei Bestand)
bis Jg. 2021: Bestellungen von Artikeln über das Online-Bestellformular
ab Jg. 2022: Lesesaal (EG)

Order via subito

Details ▾

Article ; Online: Evaluation of ultra-low input RNA sequencing for the study of human T cell transcriptome

Jingya Wang / Sadiye Amcaoglu Rieder / Jincheng Wu / Susana Hayes / Rebecca A. Halpin / Melissa de los Reyes / Yashaswi Shrestha / Roland Kolbeck / Rajiv Raja

Scientific Reports, Vol 9, Iss 1, Pp 1-

2019 Volume 13

Abstract: Abstract Deeper understanding of T cell biology is crucial for the development of new therapeutics. Human naïve T cells have low RNA content and their numbers can be limiting; therefore we set out to determine the parameters for robust ultra-low input ... ...

Abstract	Abstract Deeper understanding of T cell biology is crucial for the development of new therapeutics. Human naïve T cells have low RNA content and their numbers can be limiting; therefore we set out to determine the parameters for robust ultra-low input RNA sequencing. We performed transcriptome profiling at different cell inputs and compared three protocols: Switching Mechanism at 5′ End of RNA Template technology (SMART) with two different library preparation methods (Nextera and Clontech), and AmpliSeq technology. As the cell input decreased the number of detected coding genes decreased with SMART, while stayed constant with AmpliSeq. However, SMART enables detection of non-coding genes, which is not feasible for AmpliSeq. The detection is dependent on gene abundance, but not transcript length. The consistency between technical replicates and cell inputs was comparable across methods above 1 K but highly variable at 100 cell input. Sensitivity of detection for differentially expressed genes decreased dramatically with decreased cell inputs in all protocols, support that additional approaches, such as pathway enrichment, are important for data interpretation at ultra-low input. Finally, T cell activation signature was detected at 1 K cell input and above in all protocols, with AmpliSeq showing better detection at 100 cells.
Keywords	Medicine ; R ; Science ; Q
Subject code	612
Language	English
Publishing date	2019-06-01T00:00:00Z
Publisher	Nature Publishing Group
Document type	Article ; Online
Database	BASE - Bielefeld Academic Search Engine (life sciences selection)

Full text online

Full text

Inter-library loan at ZB MED

Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

Article ; Online: Evaluation of ultra-low input RNA sequencing for the study of human T cell transcriptome.

Wang, Jingya / Rieder, Sadiye Amcaoglu / Wu, Jincheng / Hayes, Susana / Halpin, Rebecca A / de Los Reyes, Melissa / Shrestha, Yashaswi / Kolbeck, Roland / Raja, Rajiv

Scientific reports

2019 Volume 9, Issue 1, Page(s) 8445

Abstract: Deeper understanding of T cell biology is crucial for the development of new therapeutics. Human naïve T cells have low RNA content and their numbers can be limiting; therefore we set out to determine the parameters for robust ultra-low input RNA ... ...

Abstract	Deeper understanding of T cell biology is crucial for the development of new therapeutics. Human naïve T cells have low RNA content and their numbers can be limiting; therefore we set out to determine the parameters for robust ultra-low input RNA sequencing. We performed transcriptome profiling at different cell inputs and compared three protocols: Switching Mechanism at 5' End of RNA Template technology (SMART) with two different library preparation methods (Nextera and Clontech), and AmpliSeq technology. As the cell input decreased the number of detected coding genes decreased with SMART, while stayed constant with AmpliSeq. However, SMART enables detection of non-coding genes, which is not feasible for AmpliSeq. The detection is dependent on gene abundance, but not transcript length. The consistency between technical replicates and cell inputs was comparable across methods above 1 K but highly variable at 100 cell input. Sensitivity of detection for differentially expressed genes decreased dramatically with decreased cell inputs in all protocols, support that additional approaches, such as pathway enrichment, are important for data interpretation at ultra-low input. Finally, T cell activation signature was detected at 1 K cell input and above in all protocols, with AmpliSeq showing better detection at 100 cells.
MeSH term(s)	Gene Expression Profiling ; High-Throughput Nucleotide Sequencing ; Humans ; RNA, Messenger/genetics ; Sequence Analysis, RNA/methods ; T-Lymphocytes/metabolism ; Transcriptome/genetics ; Whole Exome Sequencing
Chemical Substances	RNA, Messenger
Language	English
Publishing date	2019-06-11
Publishing country	England
Document type	Journal Article
ZDB-ID	2615211-3
ISSN	2045-2322 ; 2045-2322
ISSN (online)	2045-2322
ISSN	2045-2322
DOI	10.1038/s41598-019-44902-z
Database	MEDical Literature Analysis and Retrieval System OnLINE

Order via subito

To top

Full text online

More links

Kategorien

Order via subito

Inter-library loan at ZB MED

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

Order via subito

Inter-library loan at ZB MED

Full text online

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

Full text online

More links

Kategorien

In stock of ZB MED Cologne/Königswinter

Order via subito

Full text online

More links

Kategorien

Inter-library loan at ZB MED

More links

Kategorien

Order via subito