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

Ghosh, Santosh K.; Gerken, Thomas A.; Schneider, Keith M.; Feng, Zhihui; McCormick, Thomas S.; Weinberg, Aaron

doi:10.1373/clinchem.2006.081430

Cited by 69 publications

(57 citation statements)

References 40 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…hBD-3 peptide levels in the spent supernatants were measured by enzyme-linked immunosorbent assay (ELISA) as described previously (23). Briefly, hBD-3 antibody (Peprotech, Rocky Hill, NJ) was used to coat the bottom of the wells, followed by blocking with 1% bovine serum albumin.…”

Section: Methodsmentioning

confidence: 99%

Epithelial Innate Immune Response to Acinetobacter baumannii Challenge

et al. 2014

Self Cite

View full text Add to dashboard Cite

dCurrently, Acinetobacter baumannii is recognized as one of the major pathogens seriously threatening our health care delivery system. Aspects of the innate immune response to A. baumannii infection are not yet well understood. Human ␤-defensins (hBDs) are epithelial cell-derived cationic antimicrobial peptides (AMPs) that also function to bridge the innate and adaptive immune system. We tested the induction of hBD-2 and -3 by A. baumannii on primary oral and skin epithelial cells and found that A. baumannii induces hBD-3 transcripts to a greater extent than it induces hBD-2 transcripts on both types of cells. In addition, we found that A. baumannii is susceptible to hBD-2 and -3 killing at submicromolar concentrations. Moreover, hBD-3 induction by A. baumannii was found to be dependent on epidermal growth factor receptor (EGFR) signaling. Inhibition of mitogen-activated protein kinase resulted in reduced expression of both hBD-2 and -3. Lastly, a disintegrin and metalloprotease 17 (ADAM17; also known as TACE) was found to be critical for hBD-3 induction, while ADAM10 and dual oxidase 1 (Duox1) were not required for hBD-3 induction. Induction of AMPs is an important component of innate sensing of pathogens and may play an important role in triggering systemic immune responses to A. baumannii infection. Further studies on the interactions between epithelial cells and A. baumannii will help us understand early stages of infection and may shed light on why some individuals are more vulnerable to A. baumannii infection.

show abstract

Section: Methodsmentioning

confidence: 99%

Epithelial Innate Immune Response to Acinetobacter baumannii Challenge

et al. 2014

Self Cite

View full text Add to dashboard Cite

show abstract

“…Human beta-defensins were measured in supernatants from FnCW-stimulated HOECs following our previously published protocol (27,29).…”

mentioning

confidence: 99%

Fusobacterium nucleatum and Human Beta-Defensins Modulate the Release of Antimicrobial Chemokine CCL20/Macrophage Inflammatory Protein 3α

et al. 2011

Self Cite

View full text Add to dashboard Cite

Cells of the innate immune system regulate immune responses through the production of antimicrobial peptides, chemokines, and cytokines, including human beta-defensins (hBDs) and CCL20. In this study, we examined the kinetics of primary human oral epithelial cell (HOEC) production of CCL20 and hBDs in response to Fusobacterium nucleatum, a commensal bacterium of the oral cavity, which we previously showed promotes HOEC induction of hBDs. HOECs secrete maximal levels of CCL20 at 6 h, following stimulation by F. nucleatum cell wall (FnCW). The kinetics of CCL20 release is distinct from that of hBD-2 and -3, which peaks after 24 h and 48 h of FnCW stimulation, respectively. FnCW-induced release of CCL20 by HOECs requires both transcriptional and translational activation. Release of CCL20 by HOECs is inhibited by brefeldin A, suggesting that it is secreted through a vesicle transport pathway. Other epithelium-derived agents that FnCW induces, such as hBD-2, hBD-3, tumor necrosis factor alpha (TNF-␣) and interleukin-1␤ (IL-1␤), are also able to release CCL20. By focusing on mitogen-activated protein kinases, we show that both extracellular signalregulated kinase 1/2 and p38, but not JNK, are required for hBD-, TNF-␣-, and IL-1␤-induced secretion of CCL20 by HOECs. The ability of FnCW and its induced hBDs to produce proinflammatory cytokines and CCL20 suggests the broad role of F. nucleatum and human antimicrobial peptides in primary immune responses elicited by oral epithelium.

show abstract

“…HNP-1 to -4 are present in gingival crevicular fluid (45), and HBD2 (0.0095 g/ml) (22) and HBD3 (0.1 to 0.89 g/ml) (15,22,52) are present in saliva. Thus, HNPs and HBDs are ideally positioned in the oral cavity to interact with an extensive and diverse group of both commensal and pathogenic microbial antigens.…”

mentioning

confidence: 99%