SYNTHESIS AND SEROLOGICAL INTERACTIONS OF H.PYLORI UREASE FRAGMENT 321-339 IMMOBILIZED ON THE CELLULOSE SUPPORT

Beata Kolesińska 1Sebastian Grabowski 2Wiesław Kaca 2,3Zbigniew J. Kamiński 1

1. Technical University of Łódź, Institute of Organic Chemistry (PŁ), Żeromskiego 116, Łódź 90-924, Poland
2. University of Łódź, Department of Infectious Biology, Inst. of Microbiol. and Immunol., Łódź 90-237, Poland
3. Akademia Świętokrzyska im. Jana Kochanowskiego, Świętokrzyska 15, Kielce 25-406, Poland

Abstract

H. pylori is a major etiological agent of gastroduodenal ulcer diseases. One of the significant pathogenic factor of H. pylori is urease production and anti-urease antibodies might be responsible for inflamatory reaction proceeding atherosclerosis [1,2].

In order to study the recognitions of H.pylori UreB epitopes by sera of atherosclerosis patients we prepared 321-339 urease fragments attached to the cellulose plate with N-terminus as well as with C-terminus.

The F8 epitope: SIKEDVQF and UB-33 epitope: CHHLDKSIKEDVQFADSRI [3] were synthesized directly on the cellulose plate by using triazine based condensing reagent [4].

epitopy__1_kopia.jpg

The peptides were treated with sera of patients with medically confirmed arteriosclerosis and then with anti-human antibodies labelled by horse-radish peroxidase HRP, followed by addition 4-chloronaphtol and hydrogen peroxide.

Peptides anchored with N-terminus to cellulose gave positive reactions with patients sera more often. Results also indicated that recognition of peptides by anti-urease antibodies strongly depended on the method of bonding of epitope with cellulose plate and the structure and the length of linker between cellulose matrix and peptides.

Acknowledgement: The study was supported by the Polish State Committee for Scientific Research under the Project 4-T09A 189 25.

[1] Mendall M.A. et al., Br. Heart J., 71, 437-439 (1994).

[2] Ponzetto A. et al., BMJ, 312, 251 (1996).

[3]. Hirota K.; Nagata K., Infect. Immun., 69(11), 6597-6603 (2001).

[4] Kamiński, Z. J.; Kolesińska, B.; Kolesińska, J.; Sabatino, G.; Chelli, M.; Rovero, P.; Błaszczyk, M.; Główka, M. L.; Papini, A. M., J. Am. Chem. Soc., 127(48); 16912-16920 (2005).

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Presentation: Poster at V Multidyscyplinarna Konferencja Nauki o Leku, by Beata Kolesińska
See On-line Journal of V Multidyscyplinarna Konferencja Nauki o Leku

Submitted: 2006-02-01 14:32
Revised:   2009-06-07 00:44
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