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Infection and Immunity, May 2001, p. 3248-3254, Vol. 69, No. 5
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.5.3248-3254.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.

Calprotectin Expression by Gingival Epithelial Cells

Karen F. Ross and Mark C. Herzberg*

Department of Preventive Science, School of Dentistry, University of Minnesota, Minneapolis, Minnesota 55455

Received 26 June 2000/Returned for modification 7 September 2000/Accepted 19 January 2001

Calprotectin, a heterodimer of MRP8 and MRP14 with antimicrobial properties, is found in the cytosol of neutrophils, monocytes, and human gingival keratinocytes. During inflammation of the oral mucosa, the expression of immunoreactive calprotectin appears upregulated. Given the possible cell sources, we sought to learn if epithelial cells upregulate calprotectin in response to proinflammmatory agents. First, human gingival keratinocytes were maintained in primary culture until senescence. At each passage, cells were harvested and analyzed for quantitative expression of MRP8 and MRP14 subunit mRNA by RNase protection assays and calprotectin complex by enzyme-linked immunosorbent assay. Calprotectin expression was constitutive in the primary gingival keratinocytes, but calprotectin-specific mRNA and protein tended to increase as the cells neared senescence. To test whether calprotectin expression was inducible, immortalized gingival keratinocyte cultures were treated for 2 to 4 h with lipopolysaccharide (LPS) or interleukin-1beta (IL-1beta ). As a positive control for inducible expression, immortalized keratinocytes were incubated with phorbol myristate acetate (PMA) (50 ng/ml) for 24 h. Incubation with PMA stimulated increased expression of MRP8 and MRP14 mRNA within 2 h, peaking within 5 h. MRP8- and MRP14-specific mRNA expression by immortalized keratinocytes appeared to be unaffected by LPS or IL-1beta . In contrast, LPS, IL-1beta , and PMA each upregulated IL-8. These data show that calprotectin mRNA is expressed constitutively in cultured keratinocytes, while expression by immortalized cells appears to be independent of the exogenous proinflammatory agents LPS and IL-1beta .

* Corresponding author. Mailing address: Department of Preventive Science, School of Dentistry, University of Minnesota, 17-164 Moos Tower, 515 Delaware St., SE, Minneapolis, MN 55455-0348. Phone: (612) 625-8404. Fax: (612) 626-2651. E-mail: mcherzb{at}tc.umn.edu.

Infection and Immunity, May 2001, p. 3248-3254, Vol. 69, No. 5
0019-9567/01/$04.00+0   DOI: 10.1128/IAI.69.5.3248-3254.2001
Copyright © 2001, American Society for Microbiology. All rights reserved.


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