Inhibition of multidrug-resistant Acinetobacter baumannii by nonviral expression of hCAP-18 in a bioengineered human skin tissue.

When skin is compromised, a cascade of signals initiates the rapid repair of the epidermis to prevent fluid loss and provide defense against invading microbes. During this response, keratinocytes produce host defense peptides (HDPs) that have antimicrobial activity against a diverse set of pathogens. Using nonviral vectors we have genetically modified the novel, nontumorigenic, pathogen-free human keratinocyte progenitor cell line (NIKS) to express the human cathelicidin HDP in a tissue-specific manner. NIKS skin tissue that expresses elevated levels of cathelicidin possesses key histological features of normal epidermis and displays enhanced antimicrobial activity against bacteria in vitro. Moreover, in an in vivo infected burn wound model, this tissue results in a two log reduction in a clinical isolate of multidrug-resistant Acinetobacter baumannii. Taken together, these results suggest that this genetically engineered human tissue could be applied to burns and ulcers to counteract bacterial contamination and prevent infection.

[1]  Ji Ming Wang,et al.  Ll-37, the Neutrophil Granule–And Epithelial Cell–Derived Cathelicidin, Utilizes Formyl Peptide Receptor–Like 1 (Fprl1) as a Receptor to Chemoattract Human Peripheral Blood Neutrophils, Monocytes, and T Cells , 2000, The Journal of experimental medicine.

[2]  R. Gamelli,et al.  Severity of burn injury and sepsis determines the cytokine responses of bone marrow progenitor-derived macrophages. , 2007, The Journal of trauma.

[3]  M. Rocchi,et al.  LL-37 Protects Rats against Lethal Sepsis Caused by Gram-Negative Bacteria , 2006, Antimicrobial Agents and Chemotherapy.

[4]  F. Larcher,et al.  A cutaneous gene therapy approach to treat infection through keratinocyte‐targeted overexpression of antimicrobial peptides , 2004, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[5]  R. Gamelli,et al.  Bone Marrow Norepinephrine Mediates Development of Functionally Different Macrophages After Thermal Injury and Sepsis , 2004, Annals of surgery.

[6]  K. Cornetta,et al.  Gene transfer into humans--immunotherapy of patients with advanced melanoma, using tumor-infiltrating lymphocytes modified by retroviral gene transduction. , 1990, The New England journal of medicine.

[7]  J. Oppenheim,et al.  Alarmins: chemotactic activators of immune responses. , 2005, Current opinion in immunology.

[8]  C. Baxter,et al.  Proteolytic activity in burn wound exudates and comparison of fibrin degradation products and protease inhibitors in exudates and sera. , 1994, The Journal of burn care & rehabilitation.

[9]  A. Di Nardo,et al.  Keratinocytes store the antimicrobial peptide cathelicidin in lamellar bodies. , 2005, The Journal of investigative dermatology.

[10]  H. Sudhoff,et al.  Host defence peptides in human burns. , 2008, Burns : journal of the International Society for Burn Injuries.

[11]  I. Nagaoka,et al.  Evaluation of the effects of peptide antibiotics human β‐defensins‐1/‐2 and LL‐37 on histamine release and prostaglandin D2 production from mast cells , 2001 .

[12]  J. Calafat,et al.  Human cathelicidin, hCAP-18, is processed to the antimicrobial peptide LL-37 by extracellular cleavage with proteinase 3. , 2001, Blood.

[13]  R. Hancock,et al.  Apoptosis of airway epithelial cells: human serum sensitive induction by the cathelicidin LL-37. , 2006, American journal of respiratory cell and molecular biology.

[14]  J. Schröder.  Epithelial peptide antibiotics. , 1999, Biochemical pharmacology.

[15]  R. Gamelli,et al.  Perturbed Bone Marrow Monocyte Development Following Burn Injury and Sepsis Promote Hyporesponsive Monocytes , 2008, Journal of burn care & research : official publication of the American Burn Association.

[16]  M. Hornef,et al.  Bacterial strategies for overcoming host innate and adaptive immune responses , 2002, Nature Immunology.

[17]  Richard L. Gallo,et al.  Postsecretory Processing Generates Multiple Cathelicidins for Enhanced Topical Antimicrobial Defense1 , 2004, The Journal of Immunology.

[18]  S. Zahler,et al.  An angiogenic role for the human peptide antibiotic LL-37/hCAP-18. , 2003, The Journal of clinical investigation.

[19]  S. Andreadis.  Gene-modified tissue-engineered skin: the next generation of skin substitutes. , 2007, Advances in biochemical engineering/biotechnology.

[20]  T. Ganz,et al.  In human epidermis, β-defensin 2 is packaged in lamellar bodies , 2003 .

[21]  F. Larcher,et al.  In vitro and in vivo wound healing-promoting activities of human cathelicidin LL-37. , 2008, The Journal of investigative dermatology.

[22]  J. Larrick,et al.  Human CAP18: a novel antimicrobial lipopolysaccharide-binding protein , 1995, Infection and immunity.

[23]  V. Freedman,et al.  Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. , 1975, Proceedings of the National Academy of Sciences of the United States of America.

[24]  Ulrich Kneser,et al.  Tissue engineering of cultured skin substitutes , 2005, Journal of cellular and molecular medicine.

[25]  J. Mcgowan.  Resistance in nonfermenting gram-negative bacteria: multidrug resistance to the maximum. , 2006, The American journal of medicine.

[26]  Tomas Ganz,et al.  Endogenous antimicrobial peptides and skin infections in atopic dermatitis. , 2002, The New England journal of medicine.

[27]  Ayyalusamy Ramamoorthy,et al.  LL-37, the only human member of the cathelicidin family of antimicrobial peptides. , 2006, Biochimica et biophysica acta.

[28]  F. Götz,et al.  The Presence of Peptidoglycan O-Acetyltransferase in Various Staphylococcal Species Correlates with Lysozyme Resistance and Pathogenicity , 2006, Infection and Immunity.

[29]  F. Jacobsen,et al.  Transient cutaneous adenoviral gene therapy with human host defense peptide hCAP-18/LL-37 is effective for the treatment of burn wound infections , 2005, Gene Therapy.

[30]  V. Nizet,et al.  Antimicrobial and protease inhibitory functions of the human cathelicidin (hCAP18/LL-37) prosequence. , 2003, The Journal of investigative dermatology.

[31]  F. Grinnell,et al.  Identification of proteinase 3 as the major caseinolytic activity in acute human wound fluid. , 1998, The Journal of investigative dermatology.

[32]  Jinjuan Wang,et al.  Adoptive immunotherapy for indolent non-Hodgkin lymphoma and mantle cell lymphoma using genetically modified autologous CD20-specific T cells. , 2008, Blood.

[33]  Rebeccah L. Brown,et al.  Proteolytic activity in human burn wounds , 1997, Wound repair and regeneration : official publication of the Wound Healing Society [and] the European Tissue Repair Society.

[34]  M. Robson,et al.  Wound Infection: A Failure of Wound Healing Caused by an Imbalance of Bacteria , 1997 .

[35]  L. Rice.  Challenges in identifying new antimicrobial agents effective for treating infections with Acinetobacter baumannii and Pseudomonas aeruginosa. , 2006, Clinical infectious diseases : an official publication of the Infectious Diseases Society of America.

[36]  Henry Lin,et al.  The FASEB Journal • Research Communication Kallikrein-mediated proteolysis regulates the antimicrobial effects of cathelicidins in skin , 2022 .

[37]  M. Ståhle-Bäckdahl,et al.  The cathelicidin anti-microbial peptide LL-37 is involved in re-epithelialization of human skin wounds and is lacking in chronic ulcer epithelium. , 2003, The Journal of investigative dermatology.

[38]  M. Zasloff.  Antimicrobial peptides of multicellular organisms , 2002, Nature.

[39]  Domenico Romeo,et al.  Cathelicidins: a novel protein family with a common proregion and a variable C‐terminal antimicrobial domain , 1995, FEBS letters.

[40]  L. Meisner,et al.  Normal growth and differentiation in a spontaneously immortalized near-diploid human keratinocyte cell line, NIKS. , 2000, The Journal of investigative dermatology.

[41]  M. Pollack,et al.  Exotoxin production by clinical isolates of pseudomonas aeruginosa , 1977, Infection and immunity.

[42]  A. Mason,et al.  A standard animal burn. , 1968, The Journal of trauma.

[43]  Sheila MacNeil,et al.  Progress and opportunities for tissue-engineered skin , 2007, Nature.

[44]  R. Gamelli,et al.  New Perspectives for a New Century: Implications of Pathogen Responses for the Future of Antimicrobial Therapy , 2006, Journal of burn care & research : official publication of the American Burn Association.

[45]  D. Armstrong,et al.  The role of matrix metalloproteinases in wound healing. , 2002, Journal of the American Podiatric Medical Association.

[46]  R. Gamelli,et al.  A Novel Antibacterial Gene Transfer Treatment for Multidrug-Resistant Acinetobacter Baumannii-Induced Burn Sepsis , 2007, Journal of burn care & research : official publication of the American Burn Association.

[47]  R. Bals,et al.  The Human Host Defense Peptide LL37/hCAP Accelerates Angiogenesis in PEGT/PBT Biopolymers , 2006, Annals of plastic surgery.

[48]  I. Mellman,et al.  Plasmacytoid dendritic cells sense self-DNA coupled with antimicrobial peptide , 2007, Nature.