Clinical Chemistry AACC Online Job Center
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
QUICK SEARCH:   [advanced]


     

Clinical Chemistry 53: 757-765, 2007. First published February 22, 2007; 10.1373/clinchem.2006.081430
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow 081430.Supplemental Data
Right arrow All Versions of this Article:
clinchem.2006.081430v1
53/4/757    most recent
Right arrow Submit an electronic Letter to
the Editor about this paper
Right arrow Alert me when this article is cited
Right arrow Alert me when eLetters are posted
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Ghosh, S. K.
Right arrow Articles by Weinberg, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Ghosh, S. K.
Right arrow Articles by Weinberg, A.
Related Collections
Right arrow Clinical Immunology
Right arrow Proteomics and Protein Markers
(Clinical Chemistry. 2007;53:757-765.)
© 2007 American Association for Clinical Chemistry, Inc.

Clinical Immunology

Quantification of Human ß-Defensin-2 and -3 in Body Fluids: Application for Studies of Innate Immunity

Santosh K. Ghosh1, Thomas A. Gerken2, Keith M. Schneider1, Zhimin Feng1, Thomas S. McCormick3 and Aaron Weinberg1,a

1 Department of Biological Sciences, Case School of Dental Medicine;
2 Departments of Pediatrics and Biochemistry, Case Western Reserve University, Cleveland, OH.
3 Department of Dermatology, University Hospitals of Cleveland and Case Western Reserve University, Cleveland, OH.

aAddress correspondence to this author at: School of Dental Medicine, Case Western Reserve University, Cleveland, OH 44106-4905. Fax 216-368-0145; e-mail aaron.weinberg{at}case.edu.

Background: Human ß-defensins (hBDs) are epithelial cell-derived antimicrobial and immunoregulatory cationic peptides. Our objective was to establish an analytical tool to quantify inducible hBD-2 and -3 in body fluids.

Methods: We developed sandwich ELISAs using commercially available capture and detection antibodies and determined optimal assay conditions (with 250 mmol/L CaCl2) to overcome masking by endogenous components of body fluids. We used recombinant hBD as calibrators and for recovery testing.

Results: hBD-2 and -3 detection limits were ~75 ng/L and ~3 µg/L, respectively. Mean (SD range) values in saliva samples from healthy donors (n = 60) were 9.5 (1.2–21) µg/L for hBD-2 and 326 (50–931) µg/L for hBD-3. We did not detect hBD-3 in suction blister fluid (BF; n = 10) or bronchoalveolar lavage (BAL; n = 5) from healthy participants. We detected low hBD-2 peptide concentrations in BF and BAL, 0.16 (0.03–0.32) and 0.04 (0–0.049) µg/g total protein, respectively. We observed no correlation of hBD-2 in BF and saliva or BAL and saliva from the same person. In vaginal swabs from healthy women (n = 2), mean hBD-2 and -3 concentrations were 3.42 and 103 µg/g total protein, respectively. Cervicovaginal lavage from the same women contained mean concentrations of 1.46 and 55.5 µg/g total protein.

Conclusion: These ELISA assays can measure inducible hBD peptide concentrations in body fluids by overcoming masking effects of anionic molecules. This approach may therefore be applicable for quantifying these peptides in health and disease.






HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 2007 by the American Association for Clinical Chemistry.