Application of quantum dots as fluorescent maker in cell labeling

Quantum dots (QDs) have the potential to be used as a marker in research and supporting for medical treatment because of their unique optical and electronic properties such as size-tuneable light emission, narrow emission, high photostability, etc. With the goal of applying for biomarkers, QDs are often attached with antibodies. In order to simplify the binding process, we experimented to attach adaptor protein, namely protein A/G to CdSe/ZnS QDs covered by 3-mercaptopropionic acid (MPA). 80.7 % and 51.2 % of protein A/G at the concentration of 60 μg/mL and 20 μg/mL, respectively, conjugated with QDs in phosphate buffer saline (PBS) without supporting of N-Ethyl-N’-(3dimethylaminopropyl) carbodiimide/N-hydroxysuccinimide (EDC/NHS). After attaching to protein A/G, anti-pan T antibody could recognize and visualize Jurkat T cells. In conclusion, protein A/G was conjugated successfully on QDs and initially support for application in cell labeling.

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LIME

Journal of Experimental Medicine ◽

10.1084/jem.20031484 ◽

2003 ◽

Vol 198 (10) ◽

pp. 1453-1462 ◽

Cited By ~ 74

Author(s):

Naděžda Brdičková ◽

Tomáš Brdička ◽

Pavla Angelisová ◽

Ondrej Horváth ◽

Jiří Špička ◽

...

Keyword(s):

T Cells ◽

Lipid Rafts ◽

T Cell Activation ◽

Cell Activation ◽

Ectopic Expression ◽

Adaptor Protein ◽

Negative Regulator ◽

Membrane Rafts ◽

Cross Linking ◽

Jurkat T Cells

Lymphocyte membrane rafts contain molecules critical for immunoreceptor signaling. Here, we report identification of a new raft-associated adaptor protein LIME (Lck-interacting molecule) expressed predominantly in T lymphocytes. LIME becomes tyrosine phosphorylated after cross-linking of the CD4 or CD8 coreceptors. Phospho-LIME associates with the Src family kinase Lck and its negative regulator, Csk. Ectopic expression of LIME in Jurkat T cells results in an increase of Csk in lipid rafts, increased phosphorylation of Lck and higher Ca2+ response to CD3 stimulation. Thus, LIME appears to be involved in regulation of T cell activation by coreceptors.

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Molecular Regulation of MHC Class I Chain-Related Protein A Expression after HDAC-Inhibitor Treatment of Jurkat T Cells

The Journal of Immunology ◽

10.4049/jimmunol.179.12.8235 ◽

2007 ◽

Vol 179 (12) ◽

pp. 8235-8242 ◽

Cited By ~ 40

Author(s):

Lars Andresen ◽

Helle Jensen ◽

Marianne T. Pedersen ◽

Karen A. Hansen ◽

Søren Skov

Keyword(s):

T Cells ◽

Mhc Class I ◽

Hdac Inhibitor ◽

Protein A ◽

Molecular Regulation ◽

Class I ◽

Related Protein ◽

Jurkat T Cells ◽

Inhibitor Treatment

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RNAi down-regulation of Nck1 adaptor protein in Jurkat T cells

ScienceAsia ◽

10.2306/scienceasia1513-1874.2014.40.340 ◽

2014 ◽

Vol 40 (5) ◽

pp. 340 ◽

Cited By ~ 1

Author(s):

Pussadee Paensuwan ◽

Jatuporn Ngoenkam ◽

Donruedee Sanguansermsri ◽

Boonruang Khamsri ◽

Ichaya Yiemwattana ◽

...

Keyword(s):

T Cells ◽

Adaptor Protein ◽

Jurkat T Cells ◽

Down Regulation

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The Transmembrane Adaptor Protein Trim Regulates T Cell Receptor (Tcr) Expression and Tcr-Mediated Signaling via an Association with the Tcr ζ Chain

Journal of Experimental Medicine ◽

10.1084/jem.193.11.1269 ◽

2001 ◽

Vol 193 (11) ◽

pp. 1269-1284 ◽

Cited By ~ 31

Author(s):

Henning Kirchgessner ◽

Jes Dietrich ◽

Jeanette Scherer ◽

Pia Isomäki ◽

Vladimir Korinek ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cell Surface ◽

T Cell Receptor ◽

T Cell Activation ◽

Cell Activation ◽

Adaptor Protein ◽

Cell Receptor ◽

Jurkat T Cells ◽

Integral Component

T cell receptor (TCR)-interacting molecule (TRIM) is a recently identified transmembrane adaptor protein, which is exclusively expressed in T cells. Here we demonstrate that in mature T cells, TRIM preferentially interacts with the TCR via the TCR-ζ chains and to a lesser extent via the CD3-ε/γ heterodimer. Transient or stable overexpression of TRIM in Jurkat T cells results in enhancement of TCR expression on the cell surface and elevated induction of Ca2+ mobilization after T cell activation. TRIM-mediated upregulation of TCR expression results from inhibition of spontaneous TCR internalization and stabilization of TCR complexes on the cell surface. Collectively, our data identify TRIM as a novel integral component of the TCR complex and suggest that one function of TRIM might be to modulate the strength of signals transduced through the TCR through regulation of TCR expression on the cell surface.

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Genaration and assessment of immunomagnetic nanoparticles capable of T-cell removal

Science and Technology Development Journal - Natural Sciences ◽

10.32508/stdjns.v3i2.802 ◽

2019 ◽

Vol 3 (2) ◽

pp. 74-81

Author(s):

Kien-Quang Huynh ◽

Thuan Van Tran ◽

Thao-Suong Tran-Nguyen ◽

Kieu-Hanh Thi Ta ◽

Hieu Tran-Van

Keyword(s):

T Cells ◽

Magnetic Nanoparticles ◽

Magnetic Beads ◽

Specific Binding ◽

Protein A ◽

Allogeneic Transplantation ◽

Jurkat T Cells ◽

Hematopoietic Stem ◽

Donor Tissue ◽

Covalently Linked

Hematopoietic stem cells (HSCs) transplantation has been the potential treatment for hematopoietic disorder patients. However, once they were prescribed HSCs transplantation as the therapy, especially allogeneic transplantation, they would face Graft versus Host disease (GvHD), which causes by the presence of T cells in donor tissue. To deal with the risk of GvHD, removal T cells in donor tissue prior to transplant to recipient is extremely indispensable. Nowadays, MACS technique using immuno-magnetic nanoparticles in order to deplete T cells shows potential solution in the transplantation. In this study, we prepared immuno-magnetic nanoparticles for separation of Jurkat T cells from cell culture. Anti-Jurkat T antibodies were conjugated onto magnetic nanoparticles via recombinant protein A/G, an antibody’s Fc specific binding protein. The bonds between protein A/G and immuno-magnetic nanoparticles were covalently linked by amine groups on the surface of magnetic nanoparticles and the protein through 3-(2 pyridyldithio) propionic acid N hydroxysuccinimide ester (SPDP). Approximately 85 μg of protein A/G and 21 μg of antibody were bound to one mg of magnetic beads. The immuno-magnetic nanoparticles were capable of isolating up to 53.3% of Jurkat T cells from culture medium.

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The bright future: Imaging dynamic cellular events with quantum dots

The Biochemist ◽

10.1042/bio03203012 ◽

2010 ◽

Vol 32 (3) ◽

pp. 12-17 ◽

Cited By ~ 16

Author(s):

Andrew M. Smith ◽

Mary M. Wen ◽

Shuming Nie

Keyword(s):

Quantum Dots ◽

Light Emission ◽

Emission Spectra ◽

Semiconductor Quantum Dots ◽

Broad Absorption ◽

Light Emitting ◽

Simultaneous Excitation ◽

New Class ◽

Cellular Events ◽

Optical And Electronic Properties

Semiconductor quantum dots (QDs) are tiny light-emitting particles that have emerged as a new class of fluorescent labels for biology and medicine. Compared with traditional fluorescent probes, QDs have unique optical and electronic properties such as size-tuneable light emission, narrow and symmetric emission spectra, and broad absorption spectra that enable the simultaneous excitation of multiple fluorescence colours.

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LIME, a Novel Transmembrane Adaptor Protein, Associates with p56lck and Mediates T Cell Activation

Journal of Experimental Medicine ◽

10.1084/jem.20030232 ◽

2003 ◽

Vol 198 (10) ◽

pp. 1463-1473 ◽

Cited By ~ 63

Author(s):

Eun Mi Hur ◽

Myoungsun Son ◽

Ok-Hee Lee ◽

Young Bong Choi ◽

Changwon Park ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

T Cell Activation ◽

Transcriptional Activation ◽

Cell Activation ◽

Transmembrane Domain ◽

Adaptor Protein ◽

Jurkat T Cells ◽

Downstream Signaling ◽

Src Homology

In this study, we identify and characterize a novel transmembrane adaptor protein, designated Lck-interacting membrane protein (LIME), as a binding partner of the Lck Src homology (SH)2 domain. LIME possesses a short extracellular domain, a transmembrane domain, and a cytoplasmic tail containing five tyrosine-based motifs. The protein is primarily expressed in hematopoietic cells and lung. Interestingly, LIME expression is up-regulated by TCR stimulation and sustained up to 24 h, suggesting that LIME acts throughout the early to late stages of T cell activation. LIME is localized to membrane rafts and distributed within the T cell–APC contact site. Upon TCR stimulation of Jurkat T cells, LIME associates with Lck as a tyrosine-phosphorylated protein. Experiments using Jurkat T cells expressing CD8–LIME chimera reveal that the protein associates with phosphatidylinositol 3-kinase, Grb2, Gads, and SHP2, and activates ERK1/2 and JNK but not p38. Moreover, overexpression of LIME in Jurkat T cells induces transcriptional activation of the IL-2 promoter. Our data collectively show that LIME is a raft-associated transmembrane adaptor protein linking TCR stimuli to downstream signaling pathways via associations with Lck.

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Quantum Dots: An Emerging Tool for Point-of-Care Testing

Micromachines ◽

10.3390/mi11121058 ◽

2020 ◽

Vol 11 (12) ◽

pp. 1058

Author(s):

Suchita Singh ◽

Aksha Dhawan ◽

Sonali Karhana ◽

Madhusudan Bhat ◽

Amit Kumar Dinda

Keyword(s):

Quantum Dots ◽

Biomedical Applications ◽

Bulk Material ◽

Point Of Care ◽

Diagnostic Systems ◽

Point Of Care Testing ◽

Size Dependent ◽

Semiconductor Crystals ◽

Optical And Electronic Properties ◽

Narrow Emission

Quantum dots (QDs) are semiconductor crystals in the nanodimension having unique optical and electronic properties that differ from bulk material due to quantum mechanics. The QDs have a narrow emission peak, size-dependent emission wavelength, and broad excitation range which can be utilized for diverse biomedical applications such as molecular imaging, biosensing, and diagnostic systems. This article reviews the current developments of biomedical applications of QDs with special reference to point-of-care testing.

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Functional Cooperation between c-Cbl and Src-Like Adaptor Protein 2 in the Negative Regulation of T-Cell Receptor Signaling

Molecular and Cellular Biology ◽

10.1128/mcb.22.12.4241-4255.2002 ◽

2002 ◽

Vol 22 (12) ◽

pp. 4241-4255 ◽

Cited By ~ 37

Author(s):

Michael P. Loreto ◽

Donna M. Berry ◽

C. Jane McGlade

Keyword(s):

T Cells ◽

T Cell ◽

Tyrosine Kinases ◽

Adaptor Protein ◽

Adaptor Proteins ◽

Sh2 Domain ◽

Jurkat T Cells ◽

Activated T Cells ◽

Amino Terminal ◽

Carboxy Terminal

ABSTRACT Adaptor proteins assemble multiprotein signaling complexes, enabling the transduction of intracellular signals. While many adaptor proteins positively regulate signaling in this manner, a subgroup of adaptors function as negative regulators. Here we report the identification of a hematopoiesis-specific adaptor protein that we have designated Src-like adaptor protein 2 (SLAP-2). SLAP-2 is most closely related to SLAP and contains a Src homology 3 (SH3) domain and an SH2 domain, as well as an amino-terminal myristoylation site that mediates SLAP-2 association with membranes. Following stimulation of primary thymocytes with anti-CD3 and anti-CD28, SLAP-2 coimmunoprecipitates with tyrosine-phosphorylated c-Cbl and an unidentified protein of approximately 72 kDa. In activated Jurkat T cells, SLAP-2 also binds an additional 70-kDa phosphoprotein, identified as ZAP-70. Binding of SLAP-2 to both p72 and ZAP-70 is dependent on its SH2 domain, while c-Cbl interacts with the carboxy-terminal region. Overexpression of wild-type SLAP-2 alone or in combination with c-Cbl in Jurkat T cells leads to inhibition of T-cell antigen receptor-induced activation of nuclear factor of activated T cells. The inhibitory effect of SLAP-2 requires the carboxy-terminal c-Cbl binding region. Expression of SLAP-2 with SYK or ZAP-70 in COS cells or Jurkat T cells causes the degradation of these kinases, and SLAP-2 overexpression in Jurkat T cells reduces the surface expression of CD3. These results suggest that the mechanism of action of SLAP-2 and the related protein SLAP is to promote c-Cbl-dependent degradation of the tyrosine kinases SYK and ZAP-70 and down-regulation of CD3 at the cell surface.

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