Quantification of Human β-Defensin-2 and -3 in Body Fluids: Application for Studies of Innate Immunity
Autor: | Thomas S. McCormick, Zhimin Feng, Keith M. Schneider, Thomas A. Gerken, Aaron Weinberg, Santosh K. Ghosh |
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Rok vydání: | 2007 |
Předmět: |
Adult
Male Saliva beta-Defensins Adolescent Clinical Biochemistry Enzyme-Linked Immunosorbent Assay Peptide Cervix Uteri law.invention Blister law medicine Humans Child Defensin Aged Body fluid chemistry.chemical_classification biology medicine.diagnostic_test Biochemistry (medical) Mucins Infant Middle Aged Molecular biology Immunity Innate Body Fluids Beta defensin Bronchoalveolar lavage chemistry Child Preschool Vagina Immunology biology.protein Recombinant DNA Female Antibody Bronchoalveolar Lavage Fluid |
Zdroj: | Clinical Chemistry. 53:757-765 |
ISSN: | 1530-8561 0009-9147 |
DOI: | 10.1373/clinchem.2006.081430 |
Popis: | Background: Human β-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. |
Databáze: | OpenAIRE |
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