Vesicular LL-37 Contributes to Inflammation of the Lesional Skin of Palmoplantar Pustulosis

“Pustulosis palmaris et plantaris”, or palmoplantar pustulosis (PPP), is a chronic pustular dermatitis characterized by intraepidermal palmoplantar pustules. Although early stage vesicles (preceding the pustular phase) formed in the acrosyringium contain the antimicrobial peptides cathelicidin (hCAP-18/LL-37) and dermcidin, the details of hCAP-18/LL-37 expression in such vesicles remain unclear. The principal aim of the present study was to clarify the manner of hCAP-18/LL-37 expression in PPP vesicles and to determine whether this material contributed to subsequent inflammation of lesional skin. PPP vesicle fluid (PPP-VF) induced the expression of mRNAs encoding IL-17C, IL-8, IL-1α, and IL-1β in living skin equivalents, but the level of only IL-8 mRNA decreased significantly upon stimulation of PPP vesicle with depletion of endogenous hCAP-18/LL-37 by affinity chromatography (dep-PPP-VF). Semi-quantitative dot-blot analysis revealed higher concentrations of hCAP-18/LL-37 in PPP-VF compared to healthy sweat (2.87±0.93 µM vs. 0.09±0.09 µM). This concentration of hCAP-18/LL-37 in PPP-VF could upregulate expression of IL-17C, IL-8, IL-1α, and IL-1β at both the mRNA and protein levels. Recombinant hCAP-18 was incubated with dep-PPP-VF. Proteinase 3, which converts hCAP-18 to the active form (LL-37), was present in PPP-VF. Histopathological and immunohistochemical examination revealed that early stage vesicles contained many mononuclear cells but no polymorphonuclear cells, and the mononuclear cells were CD68-positive. The epidermis surrounding the vesicle expresses monocyte chemotactic chemokine, CCL2. In conclusion, PPP-VF contains the proteinase required for LL-37 processing and also may directly upregulate IL-8 in lesional keratinocytes, in turn contributing to the subsequent inflammation of PPP lesional skin.

[1]  K. Sayama,et al.  Eccrine Sweat Contains IL-1α, IL-1β and IL-31 and Activates Epidermal Keratinocytes as a Danger Signal , 2013, PloS one.

[2]  Hideoki Ogawa,et al.  Human antimicrobial peptide LL-37 modulates proinflammatory responses induced by cytokine milieus and double-stranded RNA in human keratinocytes. , 2013, Biochemical and biophysical research communications.

[3]  H. Iizuka,et al.  Over-expression of kallikrein related peptidases in palmoplantar pustulosis. , 2012, Journal of dermatological science.

[4]  A. Ishida-Yamamoto,et al.  Patients with palmoplantar pustulosis have increased IL‐17 and IL‐22 levels both in the lesion and serum , 2011, Experimental dermatology.

[5]  M. Vitale,et al.  Hydrogen sulfide inhibits IL-8 expression in human keratinocytes via MAP kinase signaling , 2011, Laboratory Investigation.

[6]  K. Sayama,et al.  Mite allergen is a danger signal for the skin via activation of inflammasome in keratinocytes. , 2011, The Journal of allergy and clinical immunology.

[7]  H. Hedstrand,et al.  Novel findings of Langerhans cells and interleukin‐17 expression in relation to the acrosyringium and pustule in palmoplantar pustulosis , 2010, The British journal of dermatology.

[8]  A. Ishida-Yamamoto,et al.  Acrosyringium is the main site of the vesicle/pustule formation in palmoplantar pustulosis. , 2010, Journal of Investigative Dermatology.

[9]  K. Sayama,et al.  Living skin equivalents constructed using human amnions as a matrix. , 2009, Journal of dermatological science.

[10]  P. Parren,et al.  IL-8 as Antibody Therapeutic Target in Inflammatory Diseases: Reduction of Clinical Activity in Palmoplantar Pustulosis1 , 2008, The Journal of Immunology.

[11]  K. Rabe,et al.  The antimicrobial peptide LL-37 enhances IL-8 release by human airway smooth muscle cells. , 2006, The Journal of allergy and clinical immunology.

[12]  Y. Kaneda,et al.  Microbubble-enhanced ultrasound for gene transfer into living skin equivalents. , 2005, Journal of dermatological science.

[13]  H. Tagami,et al.  Localization of IL-8 and Complement Components in Lesional Skin of Psoriasis vulgaris and Pustulosis palmaris et plantaris , 2005, Dermatology.

[14]  R. Gallo,et al.  Structure-Function Relationships among Human Cathelicidin Peptides: Dissociation of Antimicrobial Properties from Host Immunostimulatory Activities , 2005, The Journal of Immunology.

[15]  C. Garbe,et al.  Dermcidin is constitutively produced by eccrine sweat glands and is not induced in epidermal cells under inflammatory skin conditions , 2004, The British journal of dermatology.

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

[17]  R. Gallo,et al.  Neonatal Skin in Mice and Humans Expresses Increased Levels of Antimicrobial Peptides: Innate Immunity During Development of the Adaptive Response , 2003, Pediatric Research.

[18]  Takaaki Ohtake,et al.  Biology and clinical relevance of naturally occurring antimicrobial peptides. , 2002, The Journal of allergy and clinical immunology.

[19]  C. Garbe,et al.  Cathelicidin anti-microbial peptide expression in sweat, an innate defense system for the skin. , 2002, The Journal of investigative dermatology.

[20]  O. Dittrich‐Breiholz,et al.  Multiple control of interleukin‐8 gene expression , 2002, Journal of leukocyte biology.

[21]  Tomio Ogasawara,et al.  A cell-free protein synthesis system for high-throughput proteomics , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[22]  Nikolaus Blin,et al.  Dermcidin: a novel human antibiotic peptide secreted by sweat glands , 2001, Nature Immunology.

[23]  Takaaki Ohtake,et al.  Innate antimicrobial peptide protects the skin from invasive bacterial infection , 2001, Nature.

[24]  A. Quinn,et al.  Expression of the Peptide Antibiotics Human β Defensin-1 and Human β Defensin-2 in Normal Human Skin , 2001 .

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

[26]  H. Jörnvall,et al.  The human antimicrobial and chemotactic peptides LL-37 and alpha-defensins are expressed by specific lymphocyte and monocyte populations. , 2000, Blood.

[27]  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.

[28]  Y Endo,et al.  A highly efficient and robust cell-free protein synthesis system prepared from wheat embryos: plants apparently contain a suicide system directed at ribosomes. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[29]  J. Cowland,et al.  Isolation of neutrophil precursors from bone marrow for biochemical and transcriptional analysis. , 1999, Journal of immunological methods.

[30]  V. Witko-Sarsat,et al.  Proteinase 3 mRNA expression is induced in monocytes but not in neutrophils of patients with cystic fibrosis , 1999, FEBS letters.

[31]  A. Johnsen,et al.  The Human Antibacterial Cathelicidin, hCAP-18, Is Bound to Lipoproteins in Plasma* , 1999, The Journal of Biological Chemistry.

[32]  M. Ackermann,et al.  Epithelial antibiotic induced in states of disease. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[33]  J. Schröder,et al.  A peptide antibiotic from human skin , 1997, Nature.

[34]  H. Wigzell,et al.  The Expression of the Gene Coding for the Antibacterial Peptide LL-37 Is Induced in Human Keratinocytes during Inflammatory Disorders* , 1997, The Journal of Biological Chemistry.

[35]  C. Kozak,et al.  Identification of CRAMP, a Cathelin-related Antimicrobial Peptide Expressed in the Embryonic and Adult Mouse* , 1997, The Journal of Biological Chemistry.

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

[37]  J. Odeberg,et al.  FALL-39, a putative human peptide antibiotic, is cysteine-free and expressed in bone marrow and testis. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[38]  R. Falk,et al.  Reactivity of antineutrophil cytoplasmic autoantibodies with mononuclear phagocytes , 1992, Journal of leukocyte biology.

[39]  H. Müller-Hermelink,et al.  Proteinase 3, the target antigen of anticytoplasmic antibodies circulating in Wegener's granulomatosis. Immunolocalization in normal and pathologic tissues. , 1991, The American journal of pathology.

[40]  M. Uehara,et al.  The morphogenesis of pustulosis palmaris et plantaris. , 1974, Archives of dermatology.

[41]  A. Quinn,et al.  Expression of the peptide antibiotics human beta defensin-1 and human beta defensin-2 in normal human skin. , 2001, The Journal of investigative dermatology.