scholarly journals The Equine CD4+ Lymphocyte Proteome

2014 ◽  
Vol 2014 ◽  
pp. 1-4 ◽  
Author(s):  
Roxane L. Degroote ◽  
Sandra Helm ◽  
Ute Klein ◽  
Ramona Schmitt ◽  
Marius Ueffing ◽  
...  
Keyword(s):  
Reference Dataset ◽  
Effector Cells ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Key Players ◽  
Cd4 Lymphocyte ◽  
Equine Recurrent Uveitis ◽  
Cell Proteome ◽  
Spontaneous Animal Model ◽  
Proteome Expression

CD4+ T cells are key players in immunology and disease pathology, including relapsing autoimmune uveitis. Equine recurrent uveitis is the only spontaneous animal model for this disease in man. Knowledge about the CD4+ cell proteome is crucial for studies on possible changes in proteome expression of CD4+ effector cells in disease. For this purpose, we generated a reference dataset of the equine CD4+ cell proteome by sorting equine CD4+ lymphocytes followed by analysis of whole cell lysate as well as membrane protein fraction using mass spectrometry.

scholarly journals Nano-LC FTICR Tandem Mass Spectrometry for Top-Down Proteomics: Routine Baseline Unit Mass Resolution of Whole Cell Lysate Proteins up to 72 kDa

2012 ◽  
Vol 84 (5) ◽  
pp. 2111-2117 ◽  
Author(s):  
Jeremiah D. Tipton ◽  
John C. Tran ◽  
Adam D. Catherman ◽  
Dorothy R. Ahlf ◽  
Kenneth R. Durbin ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Tandem Mass Spectrometry ◽  
Mass Resolution ◽  
Unit Mass ◽  
Tandem Mass ◽  
Top Down ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Unit Mass Resolution

scholarly journals Effect of Different Adjuvants on Protection and Side-Effects Induced by Helicobacter suis Whole-Cell Lysate Vaccination

PLoS ONE ◽  
2015 ◽  
Vol 10 (6) ◽  
pp. e0131364 ◽  
Author(s):  
Iris Bosschem ◽  
Jagadeesh Bayry ◽  
Ellen De Bruyne ◽  
Kim Van Deun ◽  
Annemieke Smet ◽  
...  
Keyword(s):  
Side Effects ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate

Proteomics workflow for whole cell lysate, endosome, and lysosome fractions v2

2021 ◽  
Author(s):  
Hankum Park ◽  
Frances V Hundley ◽  
J. Wade Harper
Keyword(s):  
Sample Preparation ◽  
Whole Cell ◽  
Cell Lysates ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Ms Analysis

We present a protocol for sample preparation for LC-MS analysis of whole cell lysates and for lysosomal and endosomal fractions purified by Lyso-IP and Endo-IP. Protocols for purification of lysosomes and endosomes is provided in protocol dx.doi.org/10.17504/protocols.io.byi9puh6 using cells that express endogenously tagged TMEM192-HA and stably expressing FLAG-EEA1 as descrbed in dx.doi.org/10.17504/protocols.io.byi7puhn.

Initiation by RNA polymerase II and formation of runoff transcripts containing unblocked and unmethylated 5' termini

1983 ◽  
Vol 3 (12) ◽  
pp. 2172-2179
Author(s):  
H Ernst ◽  
W Filipowicz ◽  
A J Shatkin
Keyword(s):  
Rna Polymerase ◽  
Rna Polymerase Ii ◽  
Atp Hydrolysis ◽  
Beta Globin ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Promoter Dna ◽  
Gamma S ◽  
Made In

Transcription of cloned adenovirus, beta-globin, and retrovirus long terminal repeat DNAs in HeLa whole-cell lysate was inhibited by S-adenosylhomocysteine. However, full-length 1.7-kilobase transcripts made on adenovirus 2 late promoter DNA contained 5'-terminal GpppA, consistent with specific initiation and runoff synthesis in the absence of product methylation. Formation of runoff transcripts including retrovirus RNAs that normally contain 5'-m7GpppGmpC was not decreased by replacing GTP with non-hydrolyzable analogs, and Rous-associated virus-2 runoff products made in the presence of GTP-gamma-S contained 5'-terminal gamma-S-pppGpC. The results indicate that capping and specific transcript synthesis by RNA polymerase II are not obligatorily linked in HeLa whole-cell lysate. Accurate initiation is dependent on ATP hydrolysis, and in contrast to GTP, replacement of ATP by 5'-adenylyl-imidodiphosphate blocked specific initiation of transcripts that start with either GTP (Rous-associated virus-2, Rous-associated virus-0) or ATP (beta-globin, adenovirus).

Production and preliminary evaluation of Trypanosoma evansi HSP70 for antibody detection in Equids

2015 ◽  
Vol 60 (4) ◽  
Author(s):  
Jaideep Kumar ◽  
Ashok Chaudhury ◽  
Bidhan C. Bera ◽  
Ritesh Kumar ◽  
Rajender Kumar ◽  
...  
Keyword(s):  
Ethical Issues ◽  
Terminal Region ◽  
Antibody Detection ◽  
Preliminary Evaluation ◽  
Large Set ◽  
In Vitro Production ◽  
Serum Samples ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate

AbstractThe present immuno-diagnostic method using soluble antigens from whole cell lysate antigen for trypanosomosis have certain inherent problems like lack of standardized and reproducible antigens, as well as ethical issues due to in vivo production, that could be alleviated by in vitro production. In the present study we have identified heat shock protein 70 (HSP70) from T. evansi proteome. The nucleotide sequence of T. evansi HSP70 was 2116 bp, which encodes 690 amino acid residues. The phylogenetic analysis of T. evansi HSP70 showed that T. evansi occurred within Trypanosoma clade and is most closely related to T. brucei brucei and T. brucei gambiense, whereas T. congolense HSP70 laid in separate clade. The two partial HSP70 sequences (HSP-1 from N-terminal region and HSP-2 from C-terminal region) were expressed and evaluated as diagnostic antigens using experimentally infected equine serum samples. Both recombinant proteins detected antibody in immunoblot using serum samples from experimental infected donkeys with T. evansi. Recombinant HSP-2 showed comparable antibody response to Whole cell lysate (WCL) antigen in immunoblot and ELISA. The initial results indicated that HSP70 has potential to detect the T. evansi infection and needs further validation on large set of equine serum samples.

scholarly journals Modulation of Kv Channel Expression and Function by TCR and Costimulatory Signals during Peripheral CD4+ Lymphocyte Differentiation

2002 ◽  
Vol 196 (7) ◽  
pp. 897-909 ◽  
Author(s):  
Qing-Hua Liu ◽  
Bernd K. Fleischmann ◽  
Brian Hondowicz ◽  
Curtis C. Maier ◽  
Laurence A. Turka ◽  
...  
Keyword(s):  
T Cells ◽  
Signaling Pathways ◽  
T Cell Activation ◽  
Cell Activation ◽  
Effector Cells ◽  
Kv Channel ◽  
Kv Channels ◽  
Costimulatory Signals ◽  
Cd4 Lymphocyte ◽  
Cd4 Lymphocytes

Ionic signaling pathways, including voltage-dependent potassium (Kv) channels, are instrumental in antigen-mediated responses of peripheral T cells. However, how Kv channels cooperate with other signaling pathways involved in T cell activation and differentiation is unknown. We report that multiple Kv channels are expressed by naive CD4+ lymphocytes, and that the current amplitude and kinetics are modulated by antigen receptor–mediated stimulation and costimulatory signals. Currents expressed in naive CD4+ lymphocytes are consistent with Kv1.1, Kv1.2, Kv1.3, and Kv1.6. Effector CD4+ cells generated by optimal TCR and costimulation exhibit only Kv1.3 current, but at approximately sixfold higher levels than naive cells. CD4+ lymphocytes anergized through partial stimulation exhibit similar Kv1.1, Kv1.2, and/or Kv1.6 currents, but approximately threefold more Kv1.3 current than naive cells. To determine if Kv channels contribute to the distinct functions of naive, effector, and anergized T cells, we tested their role in immunoregulatory cytokine production. Each Kv channel is required for maximal IL-2 production by naive CD4+ lymphocytes, whereas none appears to play a role in IL-2, IL-4, or IFN-γ production by effector cells. Interestingly, Kv channels in anergized lymphocytes actively suppress IL-4 production, and these functions are consistent with a role in regulating the membrane potential and calcium signaling.

Two-dimensional blue native/SDS-PAGE analysis of whole cell lysate protein complexes of rice in response to salt stress

2016 ◽  
Vol 200 ◽  
pp. 90-101 ◽  
Author(s):  
Amenehsadat Hashemi ◽  
Javad Gharechahi ◽  
Ghorbanali Nematzadeh ◽  
Faezeh Shekari ◽  
Seyed Abdollah Hosseini ◽  
...  
Keyword(s):  
Salt Stress ◽  
Protein Complexes ◽  
Two Dimensional ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Sds Page ◽  
Blue Native

scholarly journals ESCORTing proteins directly from whole cell-lysate for single-molecule studies

2017 ◽  
Vol 535 ◽  
pp. 35-42 ◽  
Author(s):  
Shwetha Srinivasan ◽  
Jagadish P. Hazra ◽  
Gayathri S. Singaraju ◽  
Debadutta Deb ◽  
Sabyasachi Rakshit
Keyword(s):  
Single Molecule ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Single Molecule Studies

scholarly journals Altered Metabolic Phenotype of Immune Cells in a Spontaneous Autoimmune Uveitis Model

2021 ◽  
Vol 12 ◽  
Author(s):  
Claudia Barfüßer ◽  
Carmen Wiedemann ◽  
Anne L. C. Hoffmann ◽  
Sieglinde Hirmer ◽  
Cornelia A. Deeg
Keyword(s):  
T Cells ◽  
Immune Cells ◽  
Basic Research ◽  
Metabolic Phenotype ◽  
Autoimmune Uveitis ◽  
Metabolic Phenotypes ◽  
Equine Recurrent Uveitis ◽  
Spontaneous Animal Model ◽  
Glycolytic Rate ◽  
Important Disease

As one of the leading causes of blindness worldwide, uveitis is an important disease. The exact pathogenesis of autoimmune uveitis is not entirely elucidated to date. Equine recurrent uveitis (ERU) represents the only spontaneous animal model for autoimmune uveitis in humans. As the metabolism of immune cells is an emerging field in research and gains more and more significance to take part in the pathogenesis of various diseases, we conducted experiments to investigate the metabolism of immune cells of ERU cases and healthy controls. To our knowledge, the link between a deviant immunometabolism and the pathogenesis of autoimmune uveitis was not investigated so far. We showed that PBMC of ERU cases had a more active metabolic phenotype in basal state by upregulating both the oxidative phosphorylation and the glycolytic pathway. We further revealed an increased compensatory glycolytic rate of PBMC and CD4+ T cells of ERU cases under mitochondrial stress conditions. These findings are in line with metabolic alterations of immune cells in other autoimmune diseases and basic research, where it was shown that activated immune cells have an increased need of energy and molecule demand for their effector function. We demonstrated a clear difference in the metabolic phenotypes of PBMC and, more specifically, CD4+ T cells of ERU cases and controls. These findings are another important step in understanding the pathogenesis of ERU and figuratively, human autoimmune uveitis.

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