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Association of Polyphosphate (poly P) Kinases with Campylobacter jejuni Invasion and Survival in Human Epithelial Cells

Pina-Mimbela, Ruby Melisa
http://rave.ohiolink.edu/etdc/view?acc_num=osu1385978458

Abstract Details

2013, Master of Science, Ohio State University, Veterinary Preventive Medicine.
The foodborne and zoonotic bacterium Campylobacter jejuni, is one of the major causes of gastrointestinal disorders in humans worldwide. This human pathogen expresses a variety of carbohydrate, protein and lipid structures decorating its outer material (OM) which have been associated with virulence. On the other hand, inorganic polyphosphate (poly P), in many bacteria, is essential for stringent survival response and adaptation, and is considered as a virulence factor. Two kinases in C. jejuni have been associated with virulence: poly phosphate kinase 1 (PPK1) and poly phosphate kinase 2 (PPK2). PPK1 is responsible for poly P generation and PPK2 for GTP biosynthesis from poly P. GTP is a molecule involved in signaling processes as well as protein and capsule synthesis. In previous studies, we demonstrated that C. jejuni delta ppk1 shows reduced stringent survival response, adaptation, and in vivo colonization in chickens, whereas, delta ppk2, in addition to its stress tolerance defect, also displays reduced virulence properties. In the present study, the contribution of component(s) present in the outer material (OM) from the wild-type, delta ppk1 and delta ppk2 to C. jejuni invasion and intracellular survival in human epithelial cells in vitro was investigated. OM from C. jejuni wild type 81-176, delta ppk1 and delta ppk2 was extracted and fractionated into carbohydrates (lipoglycans, poly-oligosaccharides), lipids, and proteins. These fractions were tested for their contribution to C. jejuni invasion, intracellular survival and IL-8 production in INT-407 human embryonic intestinal cells. Different fractions from C. jejuni 81-176 wild type, delta ppk1 and delta ppk2 were pre-incubated with INT-407 human epithelial cells prior to infection with C. jejuni wild type. Results obtained imply that components present in the C. jejuni OM are associated to invasion and intracellular survival in INT-407 cells. Additionally, poly P kinases play a role in intracellular survival in epithelial cells. Alterations in OM from delta ppk1 and delta ppk2 are suggested to be important in C. jejuni intracellular survival in INT-407 cells. Our results suggested that OM proteins mediate C. jejuni invasion and intracellular survival to human epithelial cells whereas lipoglycans are associated with intracellular survival. In addition, alterations present in delta ppk2 lipids are associated with C. jejuni intracellular survival in INT-407 cells. On the other hand, poly- and oligo- saccharides fraction from delta ppk1 and delta ppk2 are likely to play a role in C. jejuni survival within epithelial cells. Further, we evaluated production of IL-8 in INT-407 cells by OM and its fractions from wild type, delta ppk1 and delta ppk2 strains. C. jejuni OM from wild type was not able to induce IL-8 in INT-407 cells, whereas IL-8 secretion was observed when these epithelial cells were exposed to OM from delta ppk1 and delta ppk2. Our results demonstrate that proteins, lipids, and lipoglycans present in C. jejuni OM are associated with its invasion and intracellular survival in human epithelial cells. Additionally, this study provides insights about the role of poly P in modulation of C. jejuni OM composition, thereby contributing to invasion and intracellular survival.
Gireesh Rajashekara, Dr. (Advisor)
Jordi B. Torrelles, Dr. (Committee Member)
Chang-Won Lee, Dr. (Committee Member)
122 p.
Biochemistry; Cellular Biology; Microbiology; Veterinary Services
Campylobacter jejuni; Poly P kinases; invasion; survival; epithelial cells; INT-407 cells

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Citations

  • Pina-Mimbela, R. M. (2013). Association of Polyphosphate (poly P) Kinases with Campylobacter jejuni Invasion and Survival in Human Epithelial Cells [Master's thesis, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1385978458

    APA Style (7th edition)

  • Pina-Mimbela, Ruby. Association of Polyphosphate (poly P) Kinases with Campylobacter jejuni Invasion and Survival in Human Epithelial Cells. 2013. Ohio State University, Master's thesis. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1385978458.

    MLA Style (8th edition)

  • Pina-Mimbela, Ruby. "Association of Polyphosphate (poly P) Kinases with Campylobacter jejuni Invasion and Survival in Human Epithelial Cells." Master's thesis, Ohio State University, 2013. http://rave.ohiolink.edu/etdc/view?acc_num=osu1385978458

    Chicago Manual of Style (17th edition)

osu1385978458
339
© 2013, some rights reserved.Creative Commons License Association of Polyphosphate (poly P) Kinases with Campylobacter jejuni Invasion and Survival in Human Epithelial Cells by Ruby Melisa Pina-Mimbela is licensed under a Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. Based on a work at etd.ohiolink.edu.
This open access ETD is published by The Ohio State University and OhioLINK.