Shamshul Ansari1, Junko Akada1, Yuichi Matsuo1,2, Seiji Shiota3, Yoko Kudo4, Tadayoshi Okimoto4, Kazunari Murakami4, Yoshio Yamaoka5,6,7. 1. Department of Environmental and Preventive Medicine, Oita University Faculty of Medicine, Idaigaoka, Hasama-machi, Yufu, Oita, 879-5593, Japan. 2. Department of Host-Defense Biochemistry, Institute of Tropical Medicine, Nagasaki University, Nagasaki, Japan. 3. Department of General Medicine, Almeida Memorial Hospital, Oita, Japan. 4. Department of Gastroenterology, Oita University Faculty of Medicine, Yufu, Japan. 5. Department of Environmental and Preventive Medicine, Oita University Faculty of Medicine, Idaigaoka, Hasama-machi, Yufu, Oita, 879-5593, Japan. yyamaoka@oita-u.ac.jp. 6. Global Oita Medical Advanced Research Center for Health, Idaigaoka, Hasama-machi, Yufu, Oita, 879-5593, Japan. yyamaoka@oita-u.ac.jp. 7. Department of Medicine, Gastroenterology and Hepatology Section, Baylor College of Medicine, 2002 Holcombe Blvd., Houston, TX, 77030, USA. yyamaoka@oita-u.ac.jp.
Abstract
BACKGROUND: Helicobacter pylori CagA has been found to be immuno-dominant protein and utilized for the diagnosis of the infection with cagA-positive strains. It is important to characterize the peptide epitopes capable of detecting serum anti-CagA antibodies to understand CagA immunogenicity. METHODS: Sera from 171 Japanese patients were subjected for the epitope mapping study. Eighty seven peptides were designed from the CagA consensus sequence and were used for ELISA protocol to test the serum samples. The reacting anti-CagA IgG amounts to specific peptides were measured and compared. RESULTS: The study revealed a strong reactivity of two peptides (c7-NNTEPIYAQVNKKKAGQAT and c8-AGQATSPEEPIYAQVAKKV) in H. pylori-infected group. Interestingly, these two peptides contained the well-known EPIYA-A and EPIYA-B region, respectively, which are two out of three CagA phosphorylation domains. Tyrosine-phosphorylation of these peptides reduced their reactivity in most sera. Moreover, additional peptides' mapping and chimeric-peptides' experiments indicated that the amino acids (QV and KK) accommodated in right-side flanking regions of both EPIYA-motifs were essential for their strong reactivity, whereas the third EPIYA-motif containing peptide (c12-GRSASPEPIYATIDFDEA) with differing flanking amino acids was not reactive in most cases. CONCLUSIONS: Our results suggest that the amino acid sequences constituted in the two reactive peptides are the important immunogenic regions of CagA which would be useful to develop next-generation peptide-based diagnostic assays.
BACKGROUND:Helicobacter pyloriCagA has been found to be immuno-dominant protein and utilized for the diagnosis of the infection with cagA-positive strains. It is important to characterize the peptide epitopes capable of detecting serum anti-CagA antibodies to understand CagA immunogenicity. METHODS: Sera from 171 Japanese patients were subjected for the epitope mapping study. Eighty seven peptides were designed from the CagA consensus sequence and were used for ELISA protocol to test the serum samples. The reacting anti-CagAIgG amounts to specific peptides were measured and compared. RESULTS: The study revealed a strong reactivity of two peptides (c7-NNTEPIYAQVNKKKAGQAT and c8-AGQATSPEEPIYAQVAKKV) in H. pylori-infected group. Interestingly, these two peptides contained the well-known EPIYA-A and EPIYA-B region, respectively, which are two out of three CagA phosphorylation domains. Tyrosine-phosphorylation of these peptides reduced their reactivity in most sera. Moreover, additional peptides' mapping and chimeric-peptides' experiments indicated that the amino acids (QV and KK) accommodated in right-side flanking regions of both EPIYA-motifs were essential for their strong reactivity, whereas the third EPIYA-motif containing peptide (c12-GRSASPEPIYATIDFDEA) with differing flanking amino acids was not reactive in most cases. CONCLUSIONS: Our results suggest that the amino acid sequences constituted in the two reactive peptides are the important immunogenic regions of CagA which would be useful to develop next-generation peptide-based diagnostic assays.
Authors: Christian Schumann; Kathy Triantafilou; F Maximilian Rasche; Angelika Möricke; Konstanze Vogt; Martha Triantafilou; Philipp Hahn; E Marion Schneider; Philipp M Lepper Journal: Int J Med Microbiol Date: 2006-08 Impact factor: 3.473
Authors: Angela Filomena; Anna Guenther; Hannes Planatscher; Francois Topin; Joseph She; Luca Formichella; Laurent Terradot; Markus Gerhard; Thomas O Joos; Hannelore Meyer; Nicole Schneiderhan-Marra Journal: Proteomes Date: 2017-10-03