Detection in an enzyme immunoassay of an immune response to a recombinant fragment of the 128 kilodalton protein (CagA) ofHelicobacter pylori

Xiang, Zhaoying; Bugnoli, M; Ponzetto, Antonio; Morgando, A.; Figura, Natale; Covacci, A; Petracca, Roberto; Pennatini, Carlo; Censini, Stefano; Armellini, D.; Rappuoli, Rino

doi:10.1007/bf02098460

Cited by 121 publications

(88 citation statements)

References 28 publications

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“…Detection of antibodies against HP-NAP in serum by ELISA Recombinant HP-NAP was prepared and purified, and ELISA was carried out as previously described [24][25][26][27] . Briefly, immunoplates (Nunc, Denmark) were coated with rNAP (5µg/well) and incubated overnight at 4℃.…”

Section: Screening For Seropositive Individuals With Igg Antibodies Amentioning

confidence: 99%

Detection and evaluation of antibodies against neutrophil-activating protein of Helicobacter pylori in patients with gastric cancer

Long¹,

Luo²,

Li³

et al. 2009

WJG

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AIM: To detect and evaluate the antibodies againstHelicobacter pylori (H pylori ) neutrophil-activating protein (HP-NAP) in patients with gastric cancer and other gastroduodenal diseases. METHODS:Recombinant HP-NAP was prepared from a prokaryotic expression system in Escherichia coli . Serum positivity and level of HP-NAP-specific antibodies in sera from 43 patients with gastric cancer, 28 with chronic gastritis, 28 with peptic ulcer, and 89 healthy controls were measured by rHP-NAP-based ELISA. rHP-NAP-stimulated production of interleukin-8 (IL-8) and growth-related oncogene (GROα) cytokines in the culture supernatant of SGC7901 gastric epithelial cells was also detected. RESULTS:The serum positivity and mean absorbance value of HP-NAP-specific antibodies in the gastric c a n c e r g r o u p ( 9 7.7 % a n d 1 . 0 1 ± 0 . 2 4 ) w e r e significantly higher than those in the chronic gastritis group (85.7% and 0.89 ± 0.14, P < 0.005) and healthy control group (27.7% and 0.65 ± 0.18, P < 0.001). The sensitivity and specificity of ELISA for the detection of HP-NAP-specific antibodies were 95.5% and 91.5%, respectively. HP-NAP could slightly upregulate IL-8 production in gastric epithelial cell lines but had no effect on GROα production. CONCLUSION:Infection with virulent H pylori strains secreting HP-NAP is associated with severe gastroduodenal diseases, and HP-NAP may play a role in the development of gastric carcinoma. rHP-NAPbased ELISA can be used as a new method to detect H pylori infection. The direct effect of HP-NAP on gastric epithelial cells may be limited, but HP-NAP may contribute to inflammatory response or carcinogenesis by activating neutrophils.

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Section: Screening For Seropositive Individuals With Igg Antibodies Amentioning

confidence: 99%

Detection and evaluation of antibodies against neutrophil-activating protein of Helicobacter pylori in patients with gastric cancer

Long¹,

Luo²,

Li³

et al. 2009

WJG

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“…Epidemiological and serological studies have revealed that H. pylori strains containing the CagA protein are associated with more severe disease (2) and harbor a 40-kb pathogenicity island (PAI) (3,4). The PAI encodes a bacterial type IV secretory system that secretes and translocates the CagA protein into host cells (5)(6)(7)(8), where it is phosphorylated by a host-cell kinase and causes morphological changes (7).…”

mentioning

confidence: 99%

A whole-genome microarray reveals genetic diversity among Helicobacter pylori strains

Salama

Guillemin²,

McDaniel³

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

529

446

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Helicobacter pylori colonizes the stomach of half of the world's population, causing a wide spectrum of disease ranging from asymptomatic gastritis to ulcers to gastric cancer. Although the basis for these diverse clinical outcomes is not understood, more severe disease is associated with strains harboring a pathogenicity island. To characterize the genetic diversity of more and less virulent strains, we examined the genomic content of 15 H. pylori clinical isolates by using a whole genome H. pylori DNA microarray. We found that a full 22% of H. pylori genes are dispensable in one or more strains, thus defining a minimal functional core of 1281 H. pylori genes. While the core genes encode most metabolic and cellular processes, the strain-specific genes include genes unique to H. pylori, restriction modification genes, transposases, and genes encoding cell surface proteins, which may aid the bacteria under specific circumstances during their long-term infection of genetically diverse hosts. We observed distinct patterns of the strainspecific gene distribution along the chromosome, which may result from different mechanisms of gene acquisition and loss. Among the strain-specific genes, we have found a class of candidate virulence genes identified by their coinheritance with the pathogenicity island. Helicobacter pylori is a highly host-adapted bacterial pathogen that establishes a chronic infection in the human stomach and has no known animal or environmental reservoirs (1). Epidemiological and serological studies have revealed that H. pylori strains containing the CagA protein are associated with more severe disease (2) and harbor a 40-kb pathogenicity island (PAI) (3, 4). The PAI encodes a bacterial type IV secretory system that secretes and translocates the CagA protein into host cells (5-8), where it is phosphorylated by a host-cell kinase and causes morphological changes (7). The PAI also induces IL-8 production by host cells independent of the CagA protein (9 -11). Efforts to classify H. pylori strains further by DNA fingerprinting uncovered extensive diversity (12, 13). The sequencing of two H. pylori genomes from independent strains, both containing the PAI, revealed that much of this diversity is silent at the amino acid level and thus at the functional gene level (14, 15). Here we used a H. pylori DNA microarray to examine the genomic composition of H. pylori clinical isolates containing and lacking the PAI at the level of individual genes to characterize the extent of genetic diversity between strains and to search for new candidate virulence determinants. Materials and MethodsPCR Primer Design. The elements of our microarray consisted of large (mean size, 817 base pairs; 10th percentile, 130 base pairs; 90th percentile, 1,967 base pairs) DNA fragments corresponding to unique segments of individual open reading frames (ORFs). These fragments were generated by PCRs using gene-specific primers. We aimed to include in our array the superset of ORFs from both published genomes. When an ORF was present in bo...

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“…The toxin was named vacuolating cytotoxin (VacA) because it induces vacuolation of cultured mammalian cells (11,12). Toxigenic strains of H. pylori are frequently cultured from biopsies of gastroduodenal ulcers (9,(13)(14)(15), whereas H. pylori strains harboring an altered VacA toxin gene are noncytotoxic (13)(14)(15)(16)(17)(18). Oral administration of purified VacA to mice induces gastric mucosa degeneration and inflammatory cell recruitment, characteristic of H. pylori-mediated diseases (20,21).…”

mentioning

confidence: 99%

Identification of the Minimal Intracellular Vacuolating Domain of the Helicobacter pylori Vacuolating Toxin

Willhite

Blanke

1999

Journal of Biological Chemistry

107

141

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Detection in an enzyme immunoassay of an immune response to a recombinant fragment of the 128 kilodalton protein (CagA) ofHelicobacter pylori

Cited by 121 publications

References 28 publications

Detection and evaluation of antibodies against neutrophil-activating protein of Helicobacter pylori in patients with gastric cancer

Detection and evaluation of antibodies against neutrophil-activating protein of Helicobacter pylori in patients with gastric cancer

A whole-genome microarray reveals genetic diversity among Helicobacter pylori strains

Identification of the Minimal Intracellular Vacuolating Domain of the Helicobacter pylori Vacuolating Toxin

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