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Thesis
Histatin 5 attenuates IL-8 dendritic cell response to gingivalis Hemagglutinin B
University of Iowa
Master of Science (MS), University of Iowa
Spring 2011
DOI: 10.17077/etd.uutkjswd
Abstract
<p>Histatins, a group of proteins produced by human salivary glands, have a variety of innate immune functions including the ability to: kill oral microorganisms, neutralize toxins, inactivate protease/collagenase activities, inhibit co-aggregation of oral bacteria, and inhibit lipopolysaccharide mediated activities. Hemagglutinin B (HagB), a virulence factor of the periodontal pathogen <em>Porphyromonas gingivalis, </em>induces a robust cytokine and chemokine response in human myeloid dendritic cells. In this study, I hypothesize that histatin 5 can attenuate a HagB-induced chemokine response. <strong>Objectives</strong>: To characterize an expanded cytokine and chemokine response induced in human myeloid dendritic cells by HagB, and to determine if prior incubation of HagB with histatin 5 attenuates these responses. <strong>Methods</strong>: In my first experiment, 0.040 M HagB was mixed with dilutions of histatin 5 and histatin 8 (Sigma, 0.04 to 40.0 M), incubated at 37C for 30 minutes, and added to 2 x 104 human myeloid dendritic cells (Lonza, Walkersville, MD). At 24 hours, culture media was removed, and 6 cytokines and chemokines (pg/ml) were determined in cell-free supernatants (Millipore, Billerica, MA) using the Luminex 100 IS instrument (Luminex, Austin, TX). In my second experiment, 0.040 M HagB was mixed with 40.0 M histatin 5 only (e.g., 1:1000), incubated at 37C for 30 minutes, and added to 2 x 104 human myeloid dendritic cells. At 0, 1, 2, 4, 8, 16, and 24 hours post-inoculation, culture media was removed, and 26 cytokines and chemokines (pg/ml) were determined in cell-free supernatants. <strong>Results</strong>: In both experiments, human myeloid dendritic cells incubated with HagB produced Th1, Th2, and Th17 cytokines (IL-2, IL-12(p70), IFN-, IL-3, IL-4, IL-5, , IL-15, IL-17); pro-inflammatory cytokines (IL-1, IL-1, IL-6, TNF-, IL-12(p40); anti-inflammatory v cytokines (IL-10, IL-13, IFN2); chemokines (CXCL8/IL-8, CXCL10/IP-10, CCL2/MCP-1, CCL3/MIP-1, MIP-1b, CCL11/eotaxin); and colony stimulating factors (IL-7, G-CSF, GM-CSF). Histatin 5 significantly attenuated (p < 0.05) the IL-8 response induced by HagB at 8 - 16 hours and to a lesser extent, the IL-6, GM-CSF, MCP-1, MIP-1α, MIP-1β, and TNF-α response. <strong>Conclusion: </strong>Histatin 5 is an important salivary component capable of attenuating an IL-8 response. Together with human beta defensin 3, another peptide previously shown to attenuate pro-inflammatory cytokines, histatin 5 may help control and contain oral infection and inflammation by down regulating IL-8 chemotactic response.</p>
Details
- Title: Subtitle
- Histatin 5 attenuates IL-8 dendritic cell response to gingivalis Hemagglutinin B
- Creators
- Derek Steven Borgwardt - University of Iowa
- Contributors
- Kim A. Brogden (Advisor)David R Drake (Committee Member)Paula Weistroffer (Committee Member)
- Resource Type
- Thesis
- Degree Awarded
- Master of Science (MS), University of Iowa
- Degree in
- Oral Science
- Date degree season
- Spring 2011
- Publisher
- University of Iowa
- DOI
- 10.17077/etd.uutkjswd
- Number of pages
- viii, 65 pages
- Copyright
- Copyright 2011 Derek Steven Borgwardt
- Language
- English
- Description bibliographic
- Includes bibliographical references (pages 56-65).
- Academic Unit
- Oral Pathology, Radiology and Medicine
- Record Identifier
- 9983777198802771
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