In the pathogenesis of periodontal disease, invasion of the gingival epithelium by putative pathogens appears to be critical. While many mechanisms exist to protect the mucosal surface against colonization, it is unclear if gingival epithelial cells can defend against invading bacteria. Gingival keratinocytes do express calprotectin, a cytoplasmic antibacterial protein complex. the Principal Investigator hypothesizes that calprotectin serves as an innate intraepithelial antimicrobial mechanism. To test this novel hypothesis, calprotectin will be expressed and purified and complexes will be used to determine the subunit requirements for antimicrobial activity. Normal human gingival keratinocytes will be screened for expression of calprotectin. Calprotectin-negative and calprotectin-positive normal gingival keratinocytes will be compared for resistance to colonization and invasion by putative periodontal pathogens, including Porphyromonas gingivalis and Actinobacillus actinomycetemcomitans. To show that the antimicrobial effect is restricted to calprotectin, HeLa cells will be stably transfected with calprotectin-specific cDNA and compared to control negative transfections for resistance to colonization and invasion. To demonstrate the change in innate antimicrobial activity, immortalized keratinocyte cell lines will be screened for expression of calprotectin. Specific expression of calprotectin in selected immortalized lines and normal gingival keratinocytes will be abrogated with antisense DNA and cells will then be tested for altered antimicrobial properties. These studies will define if calprotectin serves an innate defense against bacterial invasion in vitro and provide strong evidence for a fundamental role in resistance to infections.

Agency
National Institute of Health (NIH)
Institute
National Institute of Dental & Craniofacial Research (NIDCR)
Type
Research Project (R01)
Project #
5R01DE011831-03
Application #
2856655
Study Section
Oral Biology and Medicine Subcommittee 1 (OBM)
Program Officer
Lunsford, Dwayne
Project Start
1997-01-06
Project End
2000-12-31
Budget Start
1999-01-01
Budget End
1999-12-31
Support Year
3
Fiscal Year
1999
Total Cost
Indirect Cost
Name
University of Minnesota Twin Cities
Department
Pharmacology
Type
Schools of Pharmacy
DUNS #
168559177
City
Minneapolis
State
MN
Country
United States
Zip Code
55455
Khammanivong, Ali; Sorenson, Brent S; Ross, Karen F et al. (2016) Involvement of calprotectin (S100A8/A9) in molecular pathways associated with HNSCC. Oncotarget 7:14029-47
Ross, Karen F; Herzberg, Mark C (2016) Autonomous immunity in mucosal epithelial cells: fortifying the barrier against infection. Microbes Infect 18:387-398
Khammanivong, Ali; Wang, Chengxing; Sorenson, Brent S et al. (2013) S100A8/A9 (calprotectin) negatively regulates G2/M cell cycle progression and growth of squamous cell carcinoma. PLoS One 8:e69395
Bando, Mika; Zou, Xianqiong; Hiroshima, Yuka et al. (2013) Mechanism of interleukin-1? transcriptional regulation of S100A9 in a human epidermal keratinocyte cell line. Biochim Biophys Acta 1829:954-62
Sorenson, B S; Khammanivong, A; Guenther, B D et al. (2012) IL-1 receptor regulates S100A8/A9-dependent keratinocyte resistance to bacterial invasion. Mucosal Immunol 5:66-75
Hiroshima, Yuka; Bando, Mika; Kataoka, Masatoshi et al. (2011) Regulation of antimicrobial peptide expression in human gingival keratinocytes by interleukin-1?. Arch Oral Biol 56:761-7
Zhu, L; Zhang, Y; Fan, J et al. (2011) Characterization of competence and biofilm development of a Streptococcus sanguinis endocarditis isolate. Mol Oral Microbiol 26:117-26
Bando, Mika; Hiroshima, Yuka; Kataoka, Masatoshi et al. (2010) Modulation of calprotectin in human keratinocytes by keratinocyte growth factor and interleukin-1alpha. Immunol Cell Biol 88:328-33
Zaia, A A; Sappington, K J; Nisapakultorn, K et al. (2009) Subversion of antimicrobial calprotectin (S100A8/S100A9 complex) in the cytoplasm of TR146 epithelial cells after invasion by Listeria monocytogenes. Mucosal Immunol 2:43-53
Hsu, Kenneth; Champaiboon, Chantrakorn; Guenther, Brian D et al. (2009) ANTI-INFECTIVE PROTECTIVE PROPERTIES OF S100 CALGRANULINS. Antiinflamm Antiallergy Agents Med Chem 8:290-305

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