Human β-Defensins 2 and 3 Demonstrate Strain-Selective Activity against Oral Microorganisms

Joly, Sophie; Maze, Connie A.; McCray, Paul B.; Guthmiller, Janet M.

doi:10.1128/jcm.42.3.1024-1029.2004

Cited by 267 publications

(278 citation statements)

References 43 publications

(47 reference statements)

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“…Although there are available animal models and a potent hBD-2 ready to test this potentially applicable clinical treatment modality, several questions need to be addressed before a workable animal study model may be established. We stepwise tested the antimicrobial activity against C. albicans 613p that was to be used as part of the study model and verified its sensitivity to hBD-2, which has been shown to be a potent antimicrobial agent against some strains of C. albicans [6,32,33]. The effective concentrations of hBD-2 against C. albicans are 1-14 μM based on published reports, which coincides with our finding using C. albicans 613p.…”

Section: Discussionsupporting

confidence: 76%

“…1A) are in accordance with a report by Liu et al [6], which showed that E. coli is only sensitive to hBD-2 under low salt conditions. Porphyromonas gingivalis ATCC 33277 was also sensitive to hBD-2 but at a higher concentration, as reported by Joly et al [32], although it was more resistant than E. coli. Under low salt conditions, growth of P. gingivalis at 10 μM hBD-2 was completely inhibited (Fig.…”

Section: Antifungal Activities Of Hbd-2 Against C Albicans and P Gisupporting

confidence: 64%

“…We first tested its sensitivity to recombinant hBD-2 by an in vitro antimicrobial assay. The potency of hBD-2 was verified by performing parallel experiments using E. coli and P. gingivalis ATCC 33277 as positive controls, which are reported to be sensitive to hBD-2 at various concentrations [6,32].…”

Section: Antifungal Activities Of Hbd-2 Against C Albicans and P Gimentioning

confidence: 99%

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Mouse salivary glands and human β-defensin-2 as a study model for antimicrobial gene therapy: technical considerations

Yin¹,

Dang

Gazor³

et al. 2006

International Journal of Antimicrobial Agents

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Transduction of salivary glands with antimicrobial peptide genes has great potential for oral infection control. Our ultimate goal is to introduce antimicrobial peptide genes into salivary glands that secrete these peptides into saliva to control bacterial/fungal infection in the oral cavity. However, an animal study model to test this potential has not been established. Therefore, we determined to test (i) whether the potent antimicrobial peptide human β-defensin-2 (hBD-2) can be overexpressed in saliva after transduction of salivary glands and (ii) whether oral fungal infection can be developed in a NOD/SCID murine model. Lentiviral vector SIN18cPPTRhMLV bearing hBD-2 cDNA was introduced into SCID mouse submandibular glands via cannulation. Reverse transcription polymerase chain reaction (RT-PCR), immunohistochemistry or enzyme-linked immunosorbent assay (ELISA) were performed to detect hBD-2 expression in glands or in saliva. Candida albicans 613p was inoculated orally into SCID mice to establish oral candidiasis. Whilst expression of hBD-2 was detected in mouse salivary glands by RT-PCR and immunohistochemistry 1 day or 1 week following delivery of lentivirus, hBD-2 was not detected in saliva. There was recoverable C. albicans from the oral cavity and gastrointestinal tract 4 days to 4 weeks after infection, but there was no establishment of observable oral candidiasis in SCID mice under a stereomicroscope. Our data indicate that lentiviral vectors transduce mouse salivary glands, but not at a sufficient level to allow hBD-2 detection in saliva. Other vectors for gene transduction and additional treatment of SCID mice to establish oral candidiasis are needed in order to utilise mouse salivary glands to test antimicrobial gene therapy.

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Section: Discussionsupporting

confidence: 76%

Section: Antifungal Activities Of Hbd-2 Against C Albicans and P Gisupporting

confidence: 64%

Section: Antifungal Activities Of Hbd-2 Against C Albicans and P Gimentioning

confidence: 99%

See 1 more Smart Citation

Mouse salivary glands and human β-defensin-2 as a study model for antimicrobial gene therapy: technical considerations

Yin¹,

Dang

Gazor³

et al. 2006

International Journal of Antimicrobial Agents

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“…As mentioned earlier, although HBD-2 has shown antimicrobial effect on many oral and periodontal bacteria, those that play a critical role in periodontal diseases are relatively resistant to HBD-2 or require a very high concentration of HBD-2 to be killed, such as certain strains of A. actinomycetemcomitans, F. nucleatum, and P. gingivalis (MICs ranged from 6.5 to >250 µg/ml HBD-2) [64]. Sawaki et al [37] measured the HBD-2 concentration in normal oral epithelium to be ~40 µg/ml and as high as ~3.85 mg/ml in the oral squamous cell carcinoma samples.…”

Section: Discussionmentioning

confidence: 99%

Capacity of human β-defensin expression in gene-transduced and cytokine-induced cells

Yin¹,

Dang²,

Zhang³

et al. 2006

Biochemical and Biophysical Research Communications

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The purpose of this study was to determine the capacity of cells transduced with human β-defensins (HBDs) to express antimicrobial peptides, since sufficient expression level is required for effective antimicrobial activity. Retroviral vector pBabeNeo and lentiviral vector SIN18cPPTRhMLV (SIN18) carrying HBDs were utilized to transduce non-HBD-expressing cells such as fibroblasts or HBD-producing oral epithelial cells. We found that HBD-3 gene transfer to fibroblasts was possible not via retrovirus but by direct vector transfection. SIN18 had high transduction efficiencies (80.9-99.9%) and transduced cells expressed higher amounts of HBD-2 than those by pBabe-Neo. Primary human gingival epithelial cells (HGECs) expressed greater amounts of HBD-2 than primary fibroblasts after lentiviral transduction. Additionally, HBD-2 secretion from transduced HGECs cells was further increased when stimulated with IL-1 or TNFα. Our data indicate that while HBD-2 expression is limited in primary fibroblasts, its expression in HGECs may be maximized by gene transduction plus cytokine induction. KeywordsHBD-2; HBD-3; Lentiviral vectors; Retroviral vectors; IL-1; TNFα; Gingival epithelial cells Natural antimicrobial peptides have been considered as an alternative option for infection control due to the increasing resistance of microbes to conventional antibiotics. Because some potent antimicrobial peptides are encoded by single genes, the possibility of artificially delivering these genes into tissues/organs to augment innate immunity against infection or creating transgenic animals to produce antimicrobial peptides has been explored [1][2][3][4][5]. Subsequently, the term antimicrobial gene therapy has emerged [6] and engineered skin NIH-PA Author ManuscriptNIH-PA Author Manuscript NIH-PA Author Manuscript models have been used to test the concept of antimicrobial gene therapy both in vitro and in vivo [7,8]. Human β-defensins (HBDs or DEFB) and cathelicidin LL37 are being actively used as study models for their potent ability to kill a broad spectrum of bacteria and fungi [6,7,[9][10][11][12][13], as well as to inhibit viral infections [14,15]. HBDs are expressed mostly in epithelia [11,12,[16][17][18][19][20][21][22][23][24]. Besides the cloned, sequenced, and well-characterized HBD-1, -2, -3, and -4, there appears to be at least 28 more HBDs existing based on the genomic surveys using computational search strategy [25][26][27]. Fourteen HBDs or DEFB gene transcripts (DEFB-1, -4, and DEFB-103 to -114) were investigated using reverse transcriptionpolymerase chain reaction (RT-PCR) to detect their expression in gingival keratinocytes and a selected few have been classified as constitutive or inducible [27].HBDs are considered to play an important role in epithelial defense mechanism and the regulation of their expression in diseased conditions is being actively investigated. Bacteria and cytokines IL-1 and TNFα upregulate HBD-2 expression in lung and skin epithelial cells [10,28] and induction of HBDs by these stimuli is...

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“…These peptides also show strain-specific activity toward various Candida spp. (Joly et al, 2004). Interestingly, Wu and coworkers showed that hBD-3 activity against E. coli was unaffected by disulfide bonding which was previously thought to be critical to antimicrobial function .…”

mentioning

confidence: 99%

Antimicrobial Peptides in the Oral Environment: Expression and Function in Health and Disease

2005

Current Issues in Molecular Biology

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The oral cavity is a unique environment in which antimicrobial peptides play a key role in maintaining health and may have future therapeutic applications. Present evidence suggests that α-defensins, β-defensins, LL-37, histatin, and other antimicrobial peptides and proteins have distinct but overlapping roles in maintaining oral health and preventing bacterial, fungal, and viral adherence and infection. The expression of the inducible hBD-2 in normal oral epithelium, in contrast to other epithelia, and the apparent differential signaling in response to commensal and pathogenic organisms, provides new insights into innate immunity in this body site. Commensal bacteria are excellent inducers of hBD-2 in oral epithelial cells, suggesting that the commensal bacterial community acts in a manner to benefit the overall innate immune readiness of oral epithelia. This may have major significance for understanding host defense in the complex oral environment. The oral environmentThe oral cavity is a unique environment. Oral mucosa is a critical protective interface between external and internal environments and must serve as a barrier to the myriad microbial species present in this warm, moist environment. The oral cavity is the only area of the body in which hard tissues break through the epithelial surface. The periodontal epithelium surrounding the tooth is specialized to form an attachment and seal around each tooth. This unique function imparts special challenges to the tissue and leads to certain vulnerabilities associated with periodontal disease, especially in view of the continual exposure to the bacterial biofilm (dental plaque) that forms on the tooth surface at the junction of the soft tissue. Thus, this anatomical region is one where there is a significant risk of bacterially induced infection and inflammation.Antimicrobial peptides are important contributors to maintaining the balance between health and disease in this complex environment. These include several salivary antimicrobial peptides, the β-defensins expressed in the epithelium, the α-defensins expressed in neutrophils, and the cathelicidin, LL-37, expressed in both epithelium and neutrophils. These peptides are part of the host innate immune response in this environment. Epithelia, polymorphonuclear leukocytes (neutrophils), and saliva all contribute to the maintaining the health of the oral cavity in overlapping but independent ways. This review will focus on the human oral cavity and include 1) the expression and function of antimicrobial peptides in the oral cavity in the context of innate immune responses, 2) regulation of β-defensins which has led to advances in our understanding of oral epithelial innate immunity, and 3) functional efficacy against oral microbes when it is known. Epithelial antimicrobial peptidesHistorically, the oral epithelium has been considered mainly as a passive covering that becomes damaged and ulcerated in disease. This view has changed dramatically and the epithelial compartment is now seen as providing both...

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Human β-Defensins 2 and 3 Demonstrate Strain-Selective Activity against Oral Microorganisms

Cited by 267 publications

References 43 publications

Mouse salivary glands and human β-defensin-2 as a study model for antimicrobial gene therapy: technical considerations

Mouse salivary glands and human β-defensin-2 as a study model for antimicrobial gene therapy: technical considerations

Capacity of human β-defensin expression in gene-transduced and cytokine-induced cells

Antimicrobial Peptides in the Oral Environment: Expression and Function in Health and Disease

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