scholarly journals Crude Preparation of Lipopolysaccharide from Helicobacter pylori for Silver Staining and Western Blot

BIO-PROTOCOL ◽  
2017 ◽  
Vol 7 (20) ◽  
Author(s):  
Hong Li ◽  
Mohammed Benghezal
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Silver Staining ◽  
Crude Preparation

scholarly journals Production and characterization of monoclonal antibodies to the edta extract of Leptospira interrogans, serovar icterohaemorrhagiae

1996 ◽  
Vol 29 (5) ◽  
pp. 483-489
Author(s):  
Lilian Terezinha de Queiroz Leite ◽  
Mauricio Resende ◽  
Wanderley de Souza ◽  
Elizabeth R.S. Camargos ◽  
Matilde Cota Koury
Keyword(s):  
Monoclonal Antibody ◽  
Monoclonal Antibodies ◽  
Western Blot ◽  
Silver Staining ◽  
Colloidal Gold ◽  
Leptospira Interrogans ◽  
Outer Envelope ◽  
Lethal Challenge ◽  
Sds Page

Monoclonal antibodies (MABs) ivere produced against an etbylenediaminetetraacetate (EDTA) extract of Leptospira interrogans serovar icterohaemorrhagiae being characterized by gel precipitation as IgM and IgG (IgGl and IgG2b). The EDTA extract was detected as several bands by silver staining in SDS-PAGE. In the Western blot the bands around 20 KDa reacted with a monoclonal antibody, 47B4D6, and was oxidized by periodate and was not digested by pronase, suggesting that the determinant is of carbohydrate nature, lmmunocytochemistry, using colloidal gold labeling, showed that an EDTA extract determinant recognized by monoclonal antibody 47B4D6, is localized under the outer envelope of serovar icterohaemorrhagiae. Hoe AIAB raised against the EDTA extract was not able to protect hamsters from lethal challenge with virulent homologous leptospires.

scholarly journals Expression ofHelicobacter pylori hspAGene inLactococcus lactisNICE System and Experimental Study on Its Immunoreactivity

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Xiao-Juan Zhang ◽  
Shu-Ying Feng ◽  
Zhi-Tao Li ◽  
Yan-Ming Feng
Keyword(s):  
Experimental Study ◽  
Helicobacter Pylori ◽  
Western Blot ◽  
Oral Vaccine ◽  
Growth Curves ◽  
Recombinant Bacteria ◽  
Empty Plasmid ◽  
Sds Page ◽  
H Pylori ◽  
Exogenous Gene

Aim. The aim of this study was to develop an oralLactococcus lactis(L. lactis) vaccine againstHelicobacter pylori(H. pylori).Methods. AfterL. lactisNZ3900/pNZ8110-hspAwas constructed, growth curves were plotted to study whether the growth of recombinantL. lactiswas affected afterhspAwas cloned intoL. lactisand whether the growth of empty bacteria, empty plasmid bacteria, and recombinantL. lactiswas affected by different concentrations of Nisin; SDS-PAGE and Western blot were adopted, respectively, to detect the HspA expressed by recombinantL. lactisand its immunoreactivity.Results. There was no effect observed from the growth curve after exogenous genehspAwas cloned intoL. lactisNZ3900; different concentrations of Nisin did not affect the growth of NZ3900 and NZ3900/pNZ8110, while different concentrations of Nisin inhibited the growth of NZ3900/pNZ8110-hspAexcept 10 ng/mL Nisin. No HspA strip was observed from SDS-PAGE. Western blot analysis showed that HspA expressed by recombinant bacteria had favorable immunoreactivity.Conclusion. The growth of recombinantL. lactiswas suppressed even though a small amount of HspA had been induced to express. Therefore recombinantL. lactisonly express HspA which was not suitable to be oral vaccine againstHelicobacter pylori.

Evaluation of a Western Blot Test, Helico Blot 2.1, in the Diagnosis of Helicobacter pylori Infection in a Pediatric Population

Helicobacter ◽  
2002 ◽  
Vol 7 (3) ◽  
pp. 210-215 ◽  
Author(s):  
Monica Oleastro ◽  
Rita Matos ◽  
Jose Cabral ◽  
Rosa Barros ◽  
Ana Isabel Lopes ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Pediatric Population ◽  
Helicobacter Pylori Infection ◽  
Western Blot Test

scholarly journals Recombinant Helicobacter pylori CagA protein induces endoplasmic reticulum stress and autophagy in human cells

2020 ◽  
Author(s):  
Babak Nami ◽  
Ali Azzawri ◽  
Vasfiye B Ucar ◽  
Hasan Acar
Keyword(s):  
Cell Proliferation ◽  
Endoplasmic Reticulum ◽  
Cell Death ◽  
Helicobacter Pylori ◽  
Western Blot ◽  
Er Stress ◽  
Nuclear Localization ◽  
Human Cell ◽  
Human Cells ◽  
Caga Protein

AbstractHelicobacter pylori (Hp) CagA protein has a key role in the development of gastric cancer by the intruding in many intracellular processes of host human cell. Endoplasmic reticulum (ER) stress is an essential process for cellular homeostasis that modulates survival and death and is linked to several complex diseases including cancer. CagA protein is found in the serum of Hp-positive individuals and also in the supernatant of Hp culture. Limited studies report that recombinant CagA can alter gene expression and signaling pathways and induce the death of human cells. In this study, we investigated the effect of exogenous recombinant CagA protein treatment on ER stress and autophagy of human cell. AGS, MKN45, and HEK293 cells were treated with 1 µg/ml of recombinant CagA protein and then ER stress was studied by quantitative-PCR of spliced XBP-1 mRNA, immunofluorescence staining of ATF6 protein nuclear localization and real-time quantitative-PCR and/or western blot expression of GRP78, GRP94, ATF4 and CHOP genes. Autophagy was studied by western blot assessment of the conversion of LC3-I to LC3-II and LC3 aggregation. Cell proliferation and death were investigated by MTT assay and trypan blue staining respectively. As result, treatment with recombinant CagA enhanced XBP-1mRNA splicing, nuclear localization of ATF6, and the expression of ER stress signaling target genes in the cells. Recombinant CagA also induced LC3 protein conversion and aggregation in the cells. Reduced cell proliferation and increased cell death were determined in the cells treated with recombinant CagA. These results show that exogenous recombinant CagA protein causes cell death by inducing ER stress and autophagy in human cells. We conclude that CagA protein exogenously localizes in/on human cells and induces ER stress via disturbing protein machinery leading the human cell to death, however, the mechanism of CagA-host cell interaction is to be investigated.

Evaluation of a Western blot technique (Helicoblot 2.1) for the diagnosis of Helicobacter pylori infection in children

Pathology ◽  
2003 ◽  
Vol 35 (2) ◽  
pp. 157-160
Author(s):  
Dilara Öğünç ◽  
Reha Artan ◽  
Gözde Öngüt ◽  
Tekinalp Gelen ◽  
Dilek Çolak ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Helicobacter Pylori Infection ◽  
Blot Technique

scholarly journals Helicobacter pylori-Specific Immune Responses of Children: Implications for Future Vaccination Strategy

2002 ◽  
Vol 9 (5) ◽  
pp. 1126-1128 ◽  
Author(s):  
Günter Bode ◽  
Isolde Piechotowski ◽  
Dietrich Rothenbacher ◽  
Hermann Brenner
Keyword(s):  
Helicobacter Pylori ◽  
Confidence Interval ◽  
Immune Responses ◽  
Western Blot ◽  
Vaccination Strategy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Igg Antibody ◽  
Significant Difference ◽  
Specific Igg ◽  
H Pylori

ABSTRACT We analyzed the specific anti-Helicobacter pylori immunoglobulin G (IgG) antibody profile for a sample of 824 asymptomatic schoolchildren in southern Germany (mean age, 10.7 ± 0.65 years) with an H. pylori-specific IgG enzyme-linked immunosorbent assay and Western blot analysis. The prevalence of infection was 19.8% (95% confidence interval, 17.1 to 22.7%). The immunoresponses were characterized predominantly by antibodies against low-molecular-mass antigens of 14 and 29 kDa, with a significant difference between children of German and Turkish nationalities (P = 0.0012 and P < 0.0001, respectively).

scholarly journals Evaluation of Western Blot CagA Seropositivity in Helicobacter pylori-Seropositive and -Seronegative Subjects

2005 ◽  
Vol 12 (2) ◽  
pp. 304-309 ◽  
Author(s):  
J. Henrik Simán ◽  
Lars Engstrand ◽  
Göran Berglund ◽  
Claes-Henrik Florén ◽  
Arne Forsgren
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Positive Reaction ◽  
False Positive ◽  
Bimodal Distribution ◽  
Enzyme Linked Immunosorbent Assay ◽  
Birth Cohorts ◽  
False Positive Reaction ◽  
Older Subjects ◽  
H Pylori

ABSTRACT CagA seropositivity is an important risk factor for gastric adenocarcinoma and duodenal ulcer. However, CagA seropositivity is also found in Helicobacter pylori-seronegative subjects. Is CagA seropositivity in these subjects a sign of a past H. pylori infection, or does it represent a false-positive reaction? This study investigates the intensity of the CagA immune reaction and the variation in CagA seroprevalence with year of birth for 650 subjects belonging to the Malmö Preventive Medicine cohort. CagA and H. pylori seroprevalences were determined by Western blot analysis (Helicoblot 2.1) and enzyme-linked immunosorbent assay. The peak intensity of the CagA band was significantly lower in H. pylori-seronegative subjects than in those with concomitant H. pylori seropositivity. In H. pylori-seropositive subjects, peak CagA intensity had a bimodal distribution. The prevalence of CagA-seropositive but H. pylori-seronegative subjects increased successively and significantly with year of birth, in contrast to the prevalence of CagA-seropositive and H. pylori-seropositive subjects, which decreased significantly. However, within H. pylori-seropositive and -seronegative subgroups, CagA seroprevalences were constant for different birth cohorts. If CagA seropositivity in H. pylori-seronegative subjects represents a past H. pylori infection, there must have been some mechanisms of eradication that were more common in younger subjects and that were of more importance than the presence of gastric atrophy and the longer duration and higher prevalence of H. pylori infection found in older subjects. Antibiotic treatment of H. pylori was not common practice at the time of enrollment. On the other hand, a false-positive reaction would be constant and independent of birth cohorts, as with the H. pylori-seronegative subgroup of our study. Peak CagA intensity in H. pylori-seronegative subjects corresponded to the lower part of the bimodal distribution of peak CagA intensity in H. pylori-seropositive subjects. We conclude that a major proportion of CagA seropositivity in H. pylori-seronegative subjects represents a false-positive reaction. Peak CagA intensity has a bimodal distribution in H. pylori-seropositive subjects. Low-intensity CagA seropositivity in H. pylori-seropositive subjects is indeterminate, representing both false-positive and true-positive reactions.

scholarly journals Helicobacter pylori Infection and Strain Types in Adult Dyspeptic Patients Undergoing Endoscopy in a Specialized Hospital of Dhaka City

2009 ◽  
Vol 3 (1) ◽  
pp. 4-9
Author(s):  
Sufi HZ Rahman ◽  
M Anisur Rahman ◽  
MS Arfin ◽  
M Mahbub Alam ◽  
TM Bhuiyan ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Helicobacter Pylori Infection ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rapid Urease Test ◽  
Type I ◽  
Urease Test ◽  
H Pylori

Helicobacter pylori infection occurs worldwide with a high prevalence in developing countries. Virulence of H. pylori strains varies in different geographic regions. The aim of this study was to see H. pylori infection and its strain types in adult dyspeptic patients in Bangladesh and to analyze association of H. pylori strain types with clinical disease and severity of histological gastritis. Ninety consecutive adult dyspeptic patients undergoing diagnostic endoscopy were tested for H. pylori infection by culture, rapid urease test (RUT), histology and anti H. pylori IgG ELISA (Enzyme linked immunosorbent assay). H. pylori strain types were determined by Western Blot analysis. Association of strain types with clinical gastro-duodenal diseases and grades of histological gastritis were analyzed by χ2 test. Among the selected patients, 53 (58.9%) were culture positive, 48 (53.3%) were RUT positive, 31 (34.4%) were histology positive and 82 (91.1%) were anti-H. pylori IgG ELISA positive. By Western Blot analysis of the 90 sera samples, 48 (53.3%) showed antibodies to Type I strain of H. pylori, 21 (23.3%) Intermediate strain and 3 (3.3%) Type II strain. Endoscopically, 20 (22.2%) patients were found normal, 27 (30.0%) had gastritis, 9 (10.0%) had duodenitis, 28 (31.1%) had peptic ulcer disease, 4 (4.4%) had gastric carcinoma, and 2 (2.2%) had reflux esophagitis. Histologically, 34.4% had H. pylori, 44.4% had polymorhonuclear neutrophil (PMN), 100% had mononuclear cell (MNC) infiltration of different grades, 1.1% had atrophic gastritis and 2.2% had intestinal metaplasia of moderate grade. H. pylori strain types was not associated with clinical gastro-duodenal diseases or grades of PMN or MNC infiltration (p > 0.05) in these patients. Key words: Helicobacter pylori infection, H. pylori strain types, gastro-duodenal diseases, grades of gastritis   doi: 10.3329/bjmm.v3i1.2963 Bangladesh J Med Microbiol 2009; 03 (01): 4-9

scholarly journals Western Blot Method Evaluation for Detection of Helicobacter pylori Infections against H. pylori Ag in Stool Enzyme-Linked Immunoassay in Adult Egyptian Patients

2016 ◽  
Vol 4 (3) ◽  
pp. 352-358
Author(s):  
Mohamed E. Rashed ◽  
Amer M.M. ◽  
Mostafa Elnakeeb ◽  
Waleed Saeed Omer
Keyword(s):  
Helicobacter Pylori ◽  
Western Blot ◽  
Virulence Factors ◽  
Diagnostic Value ◽  
Type I ◽  
Western Blot Method ◽  
Egyptian Patients ◽  
Cytotoxin Associated Gene A ◽  
H Pylori ◽  
Medical Burden

Helicobacter pylori infection is tremendous medical burden especially in developing countries. Various immunological tests are available for diagnosis of H. pylori infection. Western blot method is proven to be promising for Precise, easy reading, sensitive and specific detection of H. pylori infections, besides it also permits the detection for the different virulence factors of CagA / VacA positive strains (type I). The objective of this study is to evaluate the diagnostic value of commercial Western Blot (WB) method in the serological diagnosis of H. pylori infections against the H. pylori Ag in stool (HpSAg) using commercial enzyme-linked immunoassay (ELISA) in adult dyspeptic Egyptian patients. Also we investigated the prevalence of virulence factors, cytotoxin-associated gene A (CagA) and vacuolating cytotoxin A (VacA) in the infected patients. Samples from 46 adult dyspeptic Egyptian patients were tested by the two methods. WB test gave accurate confirmed result with (82.6% accuracy and 89.5% sensitivity) compared to HpSAg test. Also the results indicated a high seroprevalence of cagA- and vacA-positive virulent H. pylori type I strains in adult infected population indicate that such strains may be common in this population and responsible for the majority of H. pylori infection among adult Egyptians. We concluded that WB method could be useful for the confirmatory detection of antibody profiles to H. pylori antigens and virulence factors in adult Egyptian patients.Int J Appl Sci Biotechnol, Vol 4(3): 352-358

Sign in / Sign up

Export Citation Format

Share Document