Transient cutaneous adenoviral gene therapy with human host defense peptide hCAP-18/LL-37 is effective for the treatment of burn wound infections
| Autor: | D. Mittler, Bruno Voss, Tobias Hirsch, M. Lehnhardt, A. Gerhards, Frank Jacobsen, Hans-Ulrich Steinau, Lars Steinstraesser |
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| Rok vydání: | 2005 |
| Předmět: |
Keratinocytes
Male medicine.medical_treatment Genetic Vectors Gene Expression In situ hybridization Biology Cathelicidin Adenoviridae Cell Line Rats Sprague-Dawley Western blot Transduction Genetic Cathelicidins Gene expression Genetics medicine Escherichia coli Animals Humans Transgenes Molecular Biology Cells Cultured medicine.diagnostic_test Genetic transfer Elastase Transfection Genetic Therapy Fibroblasts Molecular biology In vitro Rats Immunology Models Animal Wound Infection Molecular Medicine Burns Antimicrobial Cationic Peptides |
| Zdroj: | Gene therapy. 12(20) |
| ISSN: | 0969-7128 |
| Popis: | Host defense peptides (HDP) are naturally occurring effector molecules of the innate immune system, which might be an alternative to currently used antibiotics. The objective of this study was to investigate the efficiency of transient cutaneous adenoviral transfection with human cathelicidin hCAP-18/LL-37 in infected burn wounds. Specific transgene expression was analyzed in vitro on mRNA and protein level using real-time PCR and Western-blot. Male Sprague-Dawley rats (n=40) received a second degree scald burn on both flanks (5% BSA), which were inoculated with 10(8) colony-forming units (CFU) Pseudomonas aeruginosa. Two days later, rats were randomized into the following groups: (1) adenoviral delivery of LL-37 (Ad5-hCAP-18, n=10), (2) synthetic host defense peptide LL-37 (1 mg; n=10), (3) carrier control (PBS, n=10) and (4) empty-virus control (Ad5-LacZ, n=10). Agents were injected intradermally and subcutaneously into both flanks. After either 2 or 7 days, skin samples were harvested and homogenized. CFU per gram tissue were determined. The hCAP-18/LL-37 expression was confirmed by real-time PCR and localized using in situ hybridization. In vitro transfection of cutaneous cells delivered a specific response on mRNA production. Western blot analysis revealed protein expression of hCAP-18/LL-37 in conditioned medium and cell pellet. The host defense peptide LL-37 was detectable after cleavage of the inactive pro-form hCAP-18/LL-37 with human elastase. Ad5-hCAP-18 showed a significant bacterial inhibition of approximately 10 000 fold compared to the control group (P |
| Databáze: | OpenAIRE |
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