scholarly journals Serological Discrimination of Dogs Infected with Gastric Helicobacter spp. and Uninfected Dogs

1999 ◽  
Vol 37 (5) ◽  
pp. 1280-1287 ◽  
Author(s):  
Dalit Strauss-Ayali ◽  
Kenneth W. Simpson ◽  
Amy H. Schein ◽  
Patrick L. McDonough ◽  
Richard H. Jacobson ◽  
...  
Keyword(s):  
Molecular Mass ◽  
Bacterial Colonization ◽  
Enzyme Linked Immunosorbent Assay ◽  
Major Protein ◽  
Serum Samples ◽  
Lymphoid Follicles ◽  
Humoral Immune ◽  
Colonization Density ◽  
Antigenic Homology ◽  
H Pylori

Characterization of the humoral immune responses of people toHelicobacter pylori infection has facilitated the investigation of the host response to bacterial virulence factors and the development of sensitive and specific diagnostic tests. Dogs are commonly infected with gastric Helicobacter spp., but the presence of multiple Helicobacter spp. and possible coinfection in individual dogs have complicated serological evaluation. Evaluation of the antigenic homology of Helicobacter spp. revealed that the major protein bands of Helicobacter felisand Helicobacter bizzozeronii, two Helicobacterspp. that infect dogs, were very similar to UreA (29 to 31 kDa), UreB (63 to 66 kDa), and HSP (58 to 60 kDa) of H. pylori, and sera from infected and uninfected dogs bound in a similar way to each antigen. Immunoblotting and an enzyme-linked immunosorbent assay (ELISA) with H. felis ATCC 49179 antigen were performed with 101 serum samples (from 78 infected dogs and 23 uninfected dogs). Samples from uninfected dogs (median = 8) had fewer bands on immunoblotting than samples from infected dogs (median = 16) (P < 0.05). Combinations of the presence of any two of the low-molecular-mass bands (19, 25, 30, 32, and 37 kDa) or the high-molecular-mass bands (86 and 94 kDa) were found almost solely in samples from infected dogs (P < 0.0001). Kinetic ELISA results were significantly higher for samples from infected dogs (median = 0.0802 optical density unit [OD]/min) than for samples from uninfected dogs (median = 0.01428 OD/min). The combination of ELISA and immunoblotting results gave a specificity of 95.6% and a sensitivity of 79.8%. No correlation between ELISA results, colonization density, degree of inflammation, and presence of lymphoid follicles was observed. The results indicate substantial antigenic homology between H. felis, H. pylori, andH. bizzozeronii. The combination of ELISA and immunoblotting was a highly specific and moderately sensitive indicator of infection. The degree of seropositivity assessed by ELISA was not related to bacterial colonization density, the degree of gastric inflammation, or the presence of lymphoid follicles.

scholarly journals Quantitative Evaluation of Inflammatory and Immune Responses in the Early Stages of Chronic Helicobacter pylori Infection

2003 ◽  
Vol 71 (5) ◽  
pp. 2693-2703 ◽  
Author(s):  
Reinhard K. Straubinger ◽  
Andrea Greiter ◽  
Sean P. McDonough ◽  
Alexander Gerold ◽  
Eugenio Scanziani ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Immune Responses ◽  
Bacterial Colonization ◽  
Bacterial Virulence ◽  
Lymphoid Follicles ◽  
Early Stages ◽  
Colonization Density ◽  
Ifn Γ ◽  
H Pylori ◽  
Feline Model

ABSTRACT The early consequences of Helicobacter pylori infection and the role of bacterial virulence determinants in disease outcome remain to be established. The present study sought to measure the development of host inflammatory and immune responses and their relationship to the putative bacterial virulence factors cag pathogenicity island (cagPAI), vacA allele, and oipA in combination with bacterial colonization density in a feline model of the early stages of H. pylori infection. Gastric tissues obtained from infected and uninfected cats were evaluated for H. pylori ureB, cagPAI, vacA allele, and oipA and colonization density (urease, histology, and real-time PCR). Inflammation was assessed by measuring mRNA upregulation of gamma interferon (IFN-γ), interleukin (IL)-1α, IL-1β, IL-4, IL-6, IL-8, IL-10, and IL-12 p40 and histopathology. The mucosal immune response was characterized by morphometric analysis of lymphoid follicles and by differentiating lymphocyte populations with antibodies against surface markers. Infecting H. pylori strains were positive for vacAs1 but lacked cagPAI and an active oipA gene. Colonization density was uniform throughout the stomach. Upregulation of IFN-γ, IL-1α, IL-1β, and IL-8 and increased severity of inflammatory infiltrates and fibrosis were observed in infected cats. The median number and total area of lymphoid aggregates were 5 and 10 times greater, respectively, in the stomachs of infected than uninfected cats. Secondary lymphoid follicles in uninfected cats were rare and positive for BLA.36 and B220 but negative for CD3 and CD79α, whereas in infected cats they were frequent and positive for BLA.36, CD79α, and CD3 but negative for B220. Upregulation of IFN-γ, IL-1α, IL-1β, and IL-8 and marked hyperplasia of secondary lymphoid follicles are early consequences of H. pylori infection in cats. The response appears to be similar to that of infected people, particularly children, can develop independently of the pathogenicity factors cagPAI and oipA, and is not correlated with the degree of colonization density or urease activity.

scholarly journals Relationship of Anti-Lewis x and Anti-Lewis y Antibodies in Serum Samples from Gastric Cancer and Chronic Gastritis Patients toHelicobacter pylori-Mediated Autoimmunity

2001 ◽  
Vol 69 (8) ◽  
pp. 4774-4781 ◽  
Author(s):  
Michael A. Heneghan ◽  
Ciaran F. McCarthy ◽  
Daiva Janulaityte ◽  
Anthony P. Moran
Keyword(s):  
Gastric Cancer ◽  
Helicobacter Pylori ◽  
Gastric Mucosa ◽  
Antibody Production ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Lewis Y ◽  
H Pylori ◽  
Negative Controls ◽  
Relationship Of

ABSTRACT Lewis (Le) antigens have been implicated in the pathogenesis of atrophic gastritis and gastric cancer in the setting ofHelicobacter pylori infection, and H. pylori-induced anti-Le antibodies have been described that cross-react with the gastric mucosa of both mice and humans. The aim of this study was to examine the presence of anti-Le antibodies in patients with H. pylori infection and gastric cancer and to examine the relationships between anti-Le antibody production, bacterial Le expression, gastric histopathology, and host Le erythrocyte phenotype. Anti-Le antibody production and H. pylori Le expression were determined by enzyme-linked immunosorbent assay, erythrocyte Le phenotype was examined by agglutination assays, and histology was scored blindly. Significant levels of anti-Lex antibody (P < 0.0001, T = 76.4, DF = 5) and anti-Ley antibody (P < 0.0001, T = 73.05, DF = 5) were found in the sera of patients with gastric cancer and other H. pylori-associated pathology compared with H. pylori-negative controls. Following incubation of patient sera with synthetic Le glycoconjugates, anti-Lex and -Ley autoantibody binding was abolished. The degree of the anti-Lex and -Leyantibody response was unrelated to the host Le phenotype but was significantly associated with the bacterial expression of Lex (r = 0.863,r 2 = 0.745, P < 0.0001) and Ley (r = 0.796,r 2 = 0.634, P < 0.0001), respectively. Collectively, these data suggest that anti-Le antibodies are present in most patients with H. pyloriinfection, including those with gastric cancer, that variability exists in the strength of the anti-Le response, and that this response is independent of the host Le phenotype but related to the bacterial Le phenotype.

scholarly journals Cure of Helicobacter pylori Infection and Resolution of Gastritis by Adoptive Transfer of Splenocytes in Mice

2001 ◽  
Vol 69 (2) ◽  
pp. 1025-1031 ◽  
Author(s):  
Kathryn A. Eaton ◽  
Megan E. Mefford
Keyword(s):  
Helicobacter Pylori ◽  
Immune Responses ◽  
Adoptive Transfer ◽  
Host Response ◽  
Bacterial Colonization ◽  
Scid Mice ◽  
Humoral Immune ◽  
Cellular Immune Responses ◽  
H Pylori ◽  
Cellular Immune

ABSTRACT Vaccination suppresses Helicobacter pylori colonization but does not cure infection. Furthermore, postvaccination gastritis, likely induced by enhanced host response to residual colonization, may exacerbate disease. The goal of this study was to determine if adoptive transfer of C57BL/6 splenocytes to C57BL/6scid/scid (severe combined immunodeficient [SCID]) mice cures infection without exacerbating gastritis. H. pylori-infected and uninfected C57BL/6 mice and SCID recipients of normal splenocytes were killed at intervals between 5 and 51 weeks after infection. Colonization and gastritis were quantified, humoral immune responses were determined by enzyme-linked immunosorbent assay, and cellular immune responses were determined by delayed-type hypersensitivity response and by a proliferative response of cultured splenocytes to H. pylorisonicate. In infected C57BL/6 mice, gastritis developed gradually and bacterial colonization diminished but persisted throughout the experiment. In contrast, gastritis in infected recipient SCID mice developed rapidly and bacterial colonization decreased precipitously. Gastritis in those mice peaked 9 weeks after adoptive transfer, however, and began to resolve. By 45 weeks after transfer, gastritis had returned to background levels and bacteria were no longer detectable. Resolution of gastritis and elimination of infection were associated with a cellular but not humoral immune response to H. pylori antigens. These results demonstrate that although the host response fails to clear bacterial colonization in normal mice, enhanced cellular immune responses in recipient SCID mice are capable of clearing H. pylori infection and allowing resolution of gastritis. Thus, immune mechanisms of cure exist, and effective and safe vaccination protocols may be feasible.

scholarly journals Diversity of Antigen Recognition by Serum Antibodies in Experimental Bovine Tuberculosis

1998 ◽  
Vol 66 (11) ◽  
pp. 5344-5349 ◽  
Author(s):  
Konstantin P. Lyashchenko ◽  
John M. Pollock ◽  
Roberto Colangeli ◽  
Maria Laura Gennaro
Keyword(s):  
Antibody Production ◽  
Bovine Tuberculosis ◽  
Tuberculin Test ◽  
Economic Problem ◽  
Antigen Recognition ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Antibodies ◽  
Serum Samples ◽  
Humoral Immune ◽  
Humoral Immune Responses

ABSTRACT Tuberculosis in cattle remains a major zoonotic and economic problem in many countries. The standard diagnostic assay for bovine tuberculosis, the intradermal tuberculin test, has low accuracy. Therefore, alternative immunodiagnostic methods, such as serological assays, are needed for detection of infected animals. Development of an accurate serodiagnostic test requires a detailed understanding of the humoral immune responses during bovine tuberculosis and, in particular, identification of the key antigens of Mycobacterium bovisinvolved in antibody production. In this study, we characterized antibody responses in cattle experimentally infected with M. bovis. Sequential serum samples were collected every 3 to 4 weeks for up to 27 months postinfection. Circulating immunoglobulin G antibody levels were measured by an enzyme-linked immunosorbent assay using 12 highly purified recombinant proteins of M. bovis. Six proteins, ESAT-6, 14-kDa protein, MPT63, MPT70, MPT51, and MPT32, were identified as major seroreactive antigens in bovine tuberculosis. A remarkable animal-to-animal variation of antigen recognition by serum antibodies was observed. Kinetic analyses of the antibody production to individual antigens during infection revealed that the heterogeneous antigen recognition profile changed markedly in a given infected animal as disease progressed.

scholarly journals Immunoblot Analysis of Humoral Immune Response to Helicobacter pylori in Children with and without Duodenal Ulcer

2000 ◽  
Vol 38 (5) ◽  
pp. 1777-1781 ◽  
Author(s):  
Gifone A. Rocha ◽  
Andreia M. R. Oliveira ◽  
Dulciene M. M. Queiroz ◽  
Anfrisina S. T. Carvalho ◽  
Ana M. M. F. Nogueira
Keyword(s):  
Immune Response ◽  
Duodenal Ulcer ◽  
Helicobacter Pylori ◽  
Humoral Immune Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Specific Method ◽  
Predictive Values ◽  
Humoral Immune ◽  
Urease Test ◽  
H Pylori

Several studies have demonstrated that enzyme-linked immunosorbent assay is not a sensitive and specific method to diagnoseHelicobacter pylori infection in children, especially in the younger ones. Since serum immune response can also be determined by immunoblotting and it permits the detection of antibodies to virulence factors such as CagA and VacA, we evaluated the accuracy of a commercial immunoblotting test to diagnose H. pyloriinfection and to assess the humoral immune response to differentH. pylori antigens in 122 children who underwent upper gastrointestinal endoscopy. The presence of H. pylori was determined in antral biopsy specimens by culture, preformed urease test, and histological analysis. H. pylori was identified by microbiological and histopathological methods in 66 children (including all of the 21 who had duodenal ulcer). Antibodies toH. pylori were detected in 63 infected children and in 8 noninfected ones. The sensitivity, specificity, and positive and negative predictive values of the immunoblotting test were 95.5, 85.7, 88.7, and 94.1%, respectively. The number of immunoreactive bands increased with age (P = 0.003), and the bands of 35 kDa (P = 0.013); 89 kDa, the VacA antigen (P = 0.001); and 116 kDa, the CagA antigen (P = 0.00004) were more frequently observed in older children. The frequency of the bands of 89 kDa (P = 0.001) and 116 kDa (P = 0.03) was higher in children with duodenal ulcer than in H. pylori-positive children without the disease. In conclusion, the immunoblotting test appears to be useful for the diagnosis of H. pylori infection in children, even in the younger ones.

scholarly journals Detection of Anti-CagA Antibodies in Sera of Helicobacter pylori-Infected Patients Using an Immunochromatographic Test Strip

2019 ◽  
Vol 58 (3) ◽  
pp. 217-222
Author(s):  
Cebrail Karakus ◽  
Zeynep Ulupinar ◽  
Fahri Akbas ◽  
Duygu Yazici
Keyword(s):  
Helicobacter Pylori ◽  
Test Strip ◽  
Mucosal Damage ◽  
Enzyme Linked Immunosorbent Assay ◽  
Control Line ◽  
Immunochromatographic Test ◽  
Caga Gene ◽  
Serum Samples ◽  
Ulcer Disease ◽  
H Pylori

Abstract The cagA gene of Helicobacter pylori that encodes an immunodominant CagA protein provokes severe mucosal damage and acts as a risk factor for the development of peptic ulcer disease and gastric cancer. Our aim is to develop an immunochromatographic test strip (ICTS) using our previously developed recombinant CagA (rCagA) protein and anti-rCagA monoclonal antibody (Mab) for the detection of anti-CagA antibodies in sera of infected patients. The rCagA was firstly conjugated to gold nanoparticle and placed into the conjugate pad. A nonconjugated rCagA and anti-rCagA Mab (CK-02) were immobilized on the test line and control line, respectively. Biopsy and serum samples from 30 H. pylori-infected patients were used. The presence of cagA gene in biopsy samples was first detected by PCR (Polymerase Chain Reaction), and 22 patients were found positive while 8 were negative. When serum samples were tested by our developed ICTS, 21 were positive for anti-CagA antibodies while 9 were negative. The serum samples were also tested by a commercial ELISA (Enzyme Linked Immunosorbent Assay), and when compared to the ICTS a sensitivity of 95% and a specificity of 100% were obtained. The ICTS can be used for rapid detection of CagA-positive H. pylori infection instead of expensive, time consuming and laborious invasive approaches.

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 Use of a Novel Enzyme Immunoassay Based on Detection of Circulating Antigen in Serum for Diagnosis of Helicobacter pylori Infection

2004 ◽  
Vol 11 (4) ◽  
pp. 775-779 ◽  
Author(s):  
Abdelfattah M. Attallah ◽  
Hisham Ismail ◽  
Gellan G. Ibrahim ◽  
Mohamed Abdel-Raouf ◽  
Ahmed M. El-Waseef ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Endoscopic Biopsy ◽  
Enzyme Linked Immunosorbent Assay ◽  
Target Antigen ◽  
Serum Samples ◽  
Predictive Values ◽  
Biopsy Specimens ◽  
Significant Difference ◽  
Circulating Antigen ◽  
H Pylori

ABSTRACT Recently, noninvasive diagnostic tests for Helicobacter pylori infection have gained in significance. We have developed a sensitive and specific noninvasive immunoassay based on the detection of an H. pylori circulating antigen (HpCA) in sera from H. pylori-infected individuals. Monospecific antibody and Western blot analyses were used to demonstrate the presence of the target antigen in H. pylori cell lysate and serum samples. A novel enzyme-linked immunosorbent assay (ELISA) was developed for the detection of HpCA in serum. Endoscopic biopsy specimens from the gastric antra of 221 individuals (143 males and 78 females) with dyspeptic symptoms were evaluated for H. pylori infection, with culture used as a “gold standard” for diagnosis. The target H. pylori antigen was identified at 58 kDa. HpCA has been detected by ELISA with high degrees of sensitivity, specificity, and efficiency (>90%), and ELISA results show no significant difference (P > 0.05) from results of H. pylori culture of gastric biopsy specimens. The test's positive and negative predictive values were also high (95 and 86%, respectively). In conclusion, a sensitive and specific immunoassay was developed for the detection of HpCA in human serum. This test can be applied for noninvasive laboratory and field diagnoses of H. pylori infection.

scholarly journals Role of Helicobacter pylori cag Region Genes in Colonization and Gastritis in Two Animal Models

2001 ◽  
Vol 69 (5) ◽  
pp. 2902-2908 ◽  
Author(s):  
Kathryn A. Eaton ◽  
Dange Kersulyte ◽  
Megan Mefford ◽  
Stephen J. Danon ◽  
Steven Krakowka ◽  
...  
Keyword(s):  
Helicobacter Pylori ◽  
Bacterial Colonization ◽  
Severe Disease ◽  
Plate Count ◽  
Enzyme Linked Immunosorbent Assay ◽  
Ags Cells ◽  
Neutrophilic Inflammation ◽  
Neonatal Piglets ◽  
H Pylori

ABSTRACT The Helicobacter pylori chromosomal region known as the cytotoxin-gene associated pathogenicity island (cag PAI) is associated with severe disease and encodes proteins that are believed to induce interleukin (IL-8) secretion by cultured epithelial cells. The objective of this study was to evaluate the relationship between the cag PAI, induction of IL-8, and induction of neutrophilic gastric inflammation. Germ-free neonatal piglets and conventional C57BL/6 mice were given wild-type or cagdeficient mutant derivatives of H. pylori strain 26695 or SS1. Bacterial colonization was determined by plate count, gastritis and neutrophilic inflammation were quantified, and IL-8 induction in AGS cells was determined by enzyme-linked immunosorbent assay. Deletion of the entire cag region or interruption of thevirB10 or virB11 homolog had no effect on bacterial colonization, gastritis, or neutrophilic inflammation. In contrast, these mutations had variable effects on IL-8 induction, depending on the H. pylori strain. In the piglet-adapated strain 26695, which induced IL-8 secretion by AGS cells, deletion of the cag PAI decreased induction. In the mouse-adapted strain SS1, which did not induce IL-8 secretion, deletion of thecagII region or interruption of any of threecag region genes increased IL-8 induction. These results indicate that in mice and piglets (i) neither the cag PAI nor the ability to induce IL-8 in vitro is essential for colonization or neutrophilic inflammation and (ii) there is no direct relationship between the presence of the cag PAI, IL-8 induction, and neutrophilic gastritis.

Expression and Clinical Significance of Sperm-Associated Antigen 9 in Patients With Endometrial Carcinoma

2012 ◽  
Vol 22 (1) ◽  
pp. 87-93 ◽  
Author(s):  
Peng Yu ◽  
Lei Yan ◽  
Hui Zhang ◽  
Xiaoyan Lin ◽  
Xingbo Zhao
Keyword(s):  
Immune Response ◽  
Endometrial Cancer ◽  
Endometrial Carcinoma ◽  
Humoral Immune Response ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Humoral Immune ◽  
Significant Difference ◽  
New Type ◽  
Grade 3

ObjectiveTo investigate the expression and humoral immune response of sperm-associated antigen 9 (SPAG9) in endometri al carcinoma.MethodsSperm-associated antigen 9 gene expression levels were evaluated in endometrial carcinoma, endometrial hyperplasia, adjacent tissues, and normal endometrial tissues by reverse transcriptase-polymerase chain reaction, immunohistochemistry, and Western blot. Sperm-associated antigen 9 concentration in serum samples from 10 healthy women, 20 women with benign diseases, and 50 women with endometrial carcinoma was detected by enzyme-linked immunosorbent assay.Results(1) Sperm-associated antigen 9 antibodies were detected in approximately 72% of patients with endometrial cancer but not in healthy controls. (2) A significant difference has been found among pathological types and degrees (P < 0.05), and it was also found to be expressed in transferred lymph nodes. (3) Sperm-associated antigen 9 serum concentration (ng/mL) of patients with endometrial carcinoma is significantly higher than those of the healthy group (P < 0.05). Patients harboring grade 3 endometrial carcinoma were found to have significantly higher SPAG9 concentrations than those of grade 1/grade 2 (P = 0.003).ConclusionsSPAG9 is positively expressed in endometrial cancer, and with a high humoral immune response in patients. It may serve as a new type of endometrial cancer markers for early detection, diagnosis and treatment.

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