scholarly journals Identification of immunodominant Helicobacter pylori proteins with reactivity to H. pylori-specific egg-yolk immunoglobulin

2003 ◽  
Vol 52 (3) ◽  
pp. 217-222 ◽  
Author(s):  
Ji-Hyun Shin ◽  
Seung-Woo Nam ◽  
Jung-Taik Kim ◽  
Jong-Bok Yoon ◽  
Won-Gi Bang ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Egg Yolk ◽  
Cross Reactivity ◽  
Heat Shock Protein 60 ◽  
Α Subunit ◽  
Whole Cell ◽  
Whole Cell Lysate ◽  
Cell Lysate ◽  
Normal Human ◽  
H Pylori

The importance of hens eggs as a source of specific antibodies (IgY) is well recognized. The protective effect of IgY obtained from hens immunized with Helicobacter pylori whole-cell lysate has been reported for the control of H. pylori infection. However, IgY produced by whole-cell lysates presents the possibility of cross-reactivity with other bacteria, including the normal human flora, and this could decrease the efficiency of IgY. In the present study, the immunodominant proteins of H. pylori with reactivity to H. pylori-specific IgY (IgY-Hp) were identified. IgY obtained from hens immunized with various fractions of H. pylori proteins was isolated and purified, titres of IgY-Hp against H. pylori were determined and cross-reactivity between IgY-Hp and normal human bacteria was examined by Western blot analysis. Finally, immunodominant H. pylori proteins were identified by LC/MS analysis. IgY obtained 2 months after immunization with H. pylori whole-cell lysate showed the highest antibody titre. Five immunodominant proteins were identified that were strongly reactive to IgY-Hp: urease β-subunit (62 kDa), heat-shock protein 60 (60 kDa), urease α-subunit (26 kDa), probable peroxiredoxin (22 kDa) and probable thiol peroxidase (18 kDa). Immunization of hens with the immunodominant proteins identified would produce a more specific IgY against H. pylori.

scholarly journals Antibody to Heat Shock Protein Can Be Used for Early Serological Monitoring of Helicobacter pylori Eradication Treatment

2000 ◽  
Vol 7 (4) ◽  
pp. 574-577 ◽  
Author(s):  
Naoko Yunoki ◽  
Kenji Yokota ◽  
Motowo Mizuno ◽  
Yoshiro Kawahara ◽  
Masayasu Adachi ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Heat Shock ◽  
Antimicrobial Agents ◽  
Eradication Therapy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Peptic Ulcers ◽  
Whole Cell ◽  
Humoral Immune ◽  
Whole Cell Lysate ◽  
H Pylori

ABSTRACT Infection with Helicobacter pylori induces humoral immune responses against various antigens of the bacterium. Heat shock proteins (hsps) are immunodominant antigens in various diseases including H. pylori infection. In the present study, we measured the anti-hsp antibody titers in 42 patients with H. pylori-infected peptic ulcers during a bacterial eradication study. The patients were treated with a proton pump inhibitor and antimicrobial agents to eradicate the organism. Their sera were obtained at pretreatment and at 1 month and 6 months after the eradication therapy. The titers of immunoglobulin G antibodies to theH. pylori hsp, whole-cell lysate, and urease (30-kDa subunit) antigens in serum were measured by a capture enzyme-linked immunosorbent assay. The levels of H. pylori hsp60 antibodies in sera collected 1 month after treatment had declined significantly, even when changes in the titers of antibodies to whole-cell and urease antigens were not apparent. These results suggest that measurement of antibodies to H. pylorihsp60 in serum is useful for the early monitoring of the effectiveness of eradication therapy.

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.

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 Cross-Reactivity between Immune Responses to Helicobacter bilis and Helicobacter pylori in a Population in Thailand at High Risk of Developing Cholangiocarcinoma

2008 ◽  
Vol 15 (9) ◽  
pp. 1363-1368 ◽  
Author(s):  
Paola Pisani ◽  
Mark T. Whary ◽  
Ingrid Nilsson ◽  
Supannee Sriamporn ◽  
Torkel Wadström ◽  
...  
Keyword(s):  
High Risk ◽  
Immune Responses ◽  
Cross Reactivity ◽  
Surface Proteins ◽  
Massachusetts Institute Of Technology ◽  
Whole Cell ◽  
Serologic Tests ◽  
Helicobacter Bilis ◽  
Institute Of Technology ◽  
H Pylori

ABSTRACT Helicobacter bilis DNA has been detected in human tissue and is a candidate for etiologic investigations on the causes of hepatic and biliary tract diseases, but reliable serologic tests need to be developed in order to pursue such investigations. The scope of this study was to assess the specificity of two assays for H. bilis immune response allowing for H. pylori, and their cross-reactivity in a population in Thailand at high risk for cholangiocarcinoma. Plasma samples from 92 Thai volunteers were independently tested in two laboratories (Massachusetts Institute of Technology [MIT] and Lund). MIT performed three analyses of H. pylori and H. bilis based either on (i) outer membrane protein (OMP) with no preabsorption or on antigens derived from whole-cell sonicate before (ii) or after (iii) preabsorption with H. pylori sonicate protein. Lund used cell surface proteins from H. pylori and H. bilis as antigens. Testing for H. bilis was preabsorbed with a whole-cell lysate of H. pylori. More than 80% of the samples were positive for H. pylori in both laboratories. As tested by MIT, 58.7% (95% confidence interval, 47.9 to 68.9%) were positive for H. bilis by OMP and 44.5% (34.1 to 55.3%) were positive for H. bilis sonicate protein, but only 15.2% (8.6 to 24.2%) remained positive after preabsorption with H. pylori sonicate protein. Lund found 34.5% of the samples positive for H. bilis (22.0 to 41.0%), which was statistically compatible with all three MIT results. Serologic responses to OMPs of the two bacteria coincided in 66 and 45% of the samples in the MIT and Lund assays, respectively. We found high cross-reactivity between the immune responses to H. pylori and H. bilis antigens. More-specific H. bilis antigens need to be isolated to develop serologic tests suitable for epidemiological studies.

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