ABSTRACT
Peristomal pyoderma gangrenosum is an uncommon subtype of pyoderma gangrenosum mainly affecting stoma sites of patients with inflammatory bowel disease. While surgical treatments are often used to assist healing, little is known about the relationship between surgical interventions and the rate of recurrence of peristomal pyoderma gangrenosum. The aim of this study was to identify patient and clinical factors associated with peristomal pyoderma gangrenosum recurrence following surgical intervention. A multi-institutional retrospective case series and literature review was conducted to evaluate patient characteristics and perioperative treatment. Patients of any age with peristomal pyoderma gangrenosum undergoing surgical operations related to their pyoderma gangrenosum or due to another comorbidity were included. Descriptive statistics were used to characterize demographic information. Associations were evaluated using Wilcoxon's rank-sum test for continuous variables and Fisher's exact test for categorical data. Thirty-seven cases were included, 78.3% of which had a history of inflammatory bowel disease. Overall, 13 (35.1%) cases experienced recurrence at 30 days. There was no significant association identified between patient demographics, stoma location, surgical intervention, or perioperative treatment with rate of recurrence at 30 days post-operation. While no clinical risk factors or treatments were associated with recurrence, our work underscores the importance of a multidisciplinary approach to this disease to address gastrointestinal, dermatologic, and surgical components of treatment.
Subject(s)
Inflammatory Bowel Diseases , Pyoderma Gangrenosum , Humans , Pyoderma Gangrenosum/etiology , Pyoderma Gangrenosum/surgery , Retrospective Studies , Inflammatory Bowel Diseases/surgery , Postoperative Period , Risk FactorsABSTRACT
At present, there are no standardized guidelines for determining patient eligibility for pyoderma gangrenosum (PG) clinical trials. Thus, we aim to determine which clinical features, histopathological features, or laboratory features should be included in active ulcerative PG clinical trial eligibility criteria for treatment-naïve patients and patients already treated with immunomodulating medications (treatment-exposed patients). This study employed 4 rounds of the Delphi technique. Electronic surveys were administered to 21 international board-certified dermatologists and plastic surgeon PG experts (June 2022-December 2022). Our results demonstrated that for a patient to be eligible for a PG trial, they must meet the following criteria: (i) presence of ulcer(s) with erythematous/violaceous undermining wound borders, (ii) presence of a painful or tender ulcer, (iii) history/presence of rapidly progressing disease, (iv) exclusion of infection and other causes of cutaneous ulceration, (v) biopsy for H&E staining, and (vi) a presence/history of pathergy. These criteria vary in importance for treatment-naïve versus treatment-exposed patients. Given the international cohort, we were unable to facilitate live discussions between rounds. This Delphi consensus study provides a set of specific, standardized eligibility criteria for PG clinical trials, thus addressing one of the main issues hampering progress toward Food and Drug Administration approval of medications for PG.
Subject(s)
Clinical Trials as Topic , Consensus , Delphi Technique , Patient Selection , Pyoderma Gangrenosum , Humans , Pyoderma Gangrenosum/drug therapy , Pyoderma Gangrenosum/diagnosis , Eligibility Determination/standards , Skin Ulcer/etiology , Skin Ulcer/diagnosis , Skin Ulcer/pathology , Skin Ulcer/drug therapy , Biopsy , Skin/pathology , Skin/drug effectsABSTRACT
Importance: Chemotherapy-induced pseudocellulitis is an ill-defined term for a poorly understood phenomenon. Encompassing a myriad of cellulitis-mimicking oncologic adverse cutaneous drug reactions (ACDRs), pseudocellulitis may be difficult to diagnosis, and the lack of treatment guidance may mean unnecessary antibiotic exposure and interruptions to oncologic care. Objectives: To use case reports to characterize the various cellulitis-mimicking reactions caused by chemotherapeutic medications, to understand how these reactions affect patient care (ie, antibiotic exposure and interruptions to oncologic treatment), and to make recommendations for improved diagnosis and care of patients with chemotherapy-induced pseudocellulitis. Evidence Review: A systematic review of case reports of patients with pseudocellulitis was performed. Reports were identified through database searches using PubMed and Embase, with subsequent reference searches. Included publications described at least 1 case of chemotherapy-induced ACDR and used the term pseudocellulitis or showed evidence of cellulitis mimicry. Cases of radiation recall dermatitis were excluded. Data were extracted from a total of 32 publications representing 81 patients diagnosed with pseudocellulitis. Findings: Of the 81 cases (median [range] age, 67 [36-80] years; 44 [54%] male patients), most were associated with gemcitabine use; pemetrexed use was reported less frequently. Only 39 were considered to be true chemotherapy-induced pseudocellulitis. These cases resembled infectious cellulitis and did not meet diagnostic criteria for any known diagnoses; therefore, these were described solely as pseudocellulitis. Of this group, 26 patients (67%) had been administered antibiotics before the correct diagnosis was made, and 14 patients (36%) experienced interruptions to their oncologic treatment plans. Conclusions and Relevance: This systematic review found a variety of chemotherapy-induced ACDRs that mimic infectious cellulitis, including a group of reactions termed pseudocellulitis that do not meet criteria for other diagnoses. A more universally accepted definition and clinical research on chemotherapy-induced pseudocellulitis would allow for more accurate diagnosis, effective treatment, antibiotic stewardship, and continuation of oncologic treatment.
Subject(s)
Deoxycytidine , Drug-Related Side Effects and Adverse Reactions , Humans , Male , Aged , Female , Deoxycytidine/adverse effects , Cellulitis/diagnosis , Antimetabolites, Antineoplastic/adverse effects , Gemcitabine , Anti-Bacterial Agents/adverse effectsABSTRACT
ABSTRACT: A man with virally suppressed human immunodeficiency virus (HIV) presented with an erythematous, morbilliform rash without pustules in the setting of fever, fatigue, and myalgias after recent travel to Mexico and Puerto Rico. He was diagnosed with nonvariola orthopoxvirus (monkeypox) infection. This case report highlights an atypical presentation in the 2022 outbreak.
Subject(s)
Exanthema , HIV Infections , Mpox (monkeypox) , Male , Humans , Mpox (monkeypox)/diagnosis , Mpox (monkeypox)/epidemiology , HIV Infections/complications , HIV Infections/diagnosis , HIV Infections/epidemiology , Fever/epidemiology , Fever/etiology , Disease OutbreaksABSTRACT
Cutaneous leishmaniasis (CL) is often caused by Leishmania braziliensis (L. braziliensis) in South America. Because of the risk for mucocutaneous leishmaniasis, L. braziliensis is frequently treated with parenteral or oral medications. Here, we present a case of a young woman with L. braziliensis (CL) that did not respond to miltefosine but eventually experienced spontaneous resolution. This case highlights the potential for treatment failure and the importance of clinical monitoring in the setting of cutaneous leishmaniasis caused by L. braziliensis.
Subject(s)
Antiprotozoal Agents/therapeutic use , Leishmaniasis, Cutaneous/drug therapy , Phosphorylcholine/analogs & derivatives , Treatment Failure , Female , Humans , Leishmania braziliensis , Leishmaniasis, Cutaneous/epidemiology , Peru/epidemiology , Phosphorylcholine/therapeutic use , Young AdultABSTRACT
Tumor necrosis factor (TNF)-α inhibitors increase susceptibility to tuberculosis, but the effect of biologics on susceptibility to leprosy has not been described. Moreover, biologics may play a role in treating erythema nodosum leprosum (ENL). The objectives of this systematic review were to determine whether the development of clinical leprosy is increased in patients being treated with biologics and to assess the use of biologics in treating leprosy reactions. A systematic literature review was completed of patients with leprosy who received treatment with biologics either before or after a diagnosis of leprosy was confirmed. All studies and case reports were included for qualitative evaluation. The search yielded 10 cases (including one duplicate publication) of leprosy diagnosed after initiation of TNF-α inhibitors and four case reports of refractory ENL successfully treated with infliximab or etanercept. An unpublished case of persistent ENL responsive to infliximab is also presented. These data demonstrate that the use of TNF-α inhibitors may be a risk factor for developing leprosy or reactivating subclinical infections. Leprosy can present with skin lesions and arthritis, so leprosy should be considered in patients presenting with these signs before starting treatment with these agents. Leprosy should be considered in patients who develop worsening eruptions and neurologic symptoms during treatment with TNF-α inhibitors. Finally, TNF-α inhibitors appear effective in some cases of refractory ENL.
Subject(s)
Biological Products/therapeutic use , Leprostatic Agents/therapeutic use , Leprosy/drug therapy , Adult , Humans , Infliximab/therapeutic use , Male , Tumor Necrosis Factor-alpha/antagonists & inhibitorsABSTRACT
BACKGROUND: Leprosy, a chronic granulomatous disease affecting the skin and nerves, is caused by Mycobacterium leprae (M. leprae). The type of leprosy developed depends upon the host immune response. Type 1 reactions (T1Rs), that complicate borderline and lepromatous leprosy, are due to an increase in cell-mediated immunity and manifest as nerve damage and skin inflammation. Owing to the increase in inflammation in the skin of patients with T1Rs, we sought to investigate the activation of the innate immune system during reactionary events. Specifically, we investigated the expression levels of human beta-defensins (hBDs) 2 and 3 in the skin of patients with T1Rs, in keratinocytes, and in macrophages stimulated with M. leprae and corticosteroids. RESULTS: Skin biopsies from twenty-three patients with Type 1 reactions were found to have higher transcript levels of hBD3 as compared to fifteen leprosy patients without Type 1 reactions, as measured by qPCR. Moreover, we observed that keratinocytes but not macrophages up-regulated hBD2 and hBD3 in response to M. leprae stimulation in vitro. Corticosteroid treatment of patients with T1Rs caused a suppression of hBD2 and hBD3 in skin biopsies, as measured by qPCR. In vitro, corticosteroids suppressed M. leprae-dependent induction of hBD2 and hBD3 in keratinocytes. CONCLUSIONS: This study demonstrates that hBD3 is induced in leprosy Type 1 Reactions and suppressed by corticosteroids. Furthermore, our findings demonstrate that keratinocytes are responsive to M. leprae and lend support for additional studies on keratinocyte innate immunity in leprosy and T1Rs. TRIAL REGISTRATION: Controlled-Trials.com ISRCTN31894035.
Subject(s)
Leprosy, Lepromatous/immunology , Mycobacterium leprae/immunology , Up-Regulation , beta-Defensins/biosynthesis , Adrenal Cortex Hormones/therapeutic use , Biopsy , Cells, Cultured , Gene Expression Profiling , Humans , Immunosuppressive Agents/therapeutic use , Keratinocytes/immunology , Keratinocytes/microbiology , Macrophages/immunology , Macrophages/microbiology , Real-Time Polymerase Chain ReactionABSTRACT
Mast cells (MCs) are well-known effectors of allergic reactions and are considered sentinels in the skin and mucosa. In addition, through their production of cathelicidin, MCs have the capacity to oppose invading pathogens. We therefore hypothesized that MCs could act as sentinels in the skin against viral infections using antimicrobial peptides. In this study, we demonstrate that MCs react to vaccinia virus (VV) and degranulate using a membrane-activated pathway that leads to antimicrobial peptide discharge and virus inactivation. This finding was supported using a mouse model of viral infection. MC-deficient (Kit(wsh-/-)) mice were more susceptible to skin VV infection than the wild type animals, whereas Kit(wsh-/-) mice reconstituted with MCs in the skin showed a normal response to VV. Using MCs derived from mice deficient in cathelicidin antimicrobial peptide, we showed that antimicrobial peptides are one important antiviral granule component in in vivo skin infections. In conclusion, we demonstrate that MC presence protects mice from VV skin infection, MC degranulation is required for protecting mice from VV, neutralizing Ab to the L1 fusion entry protein of VV inhibits degranulation apparently by preventing S1PR2 activation by viral membrane lipids, and antimicrobial peptide release from MC granules is necessary to inactivate VV infectivity.
Subject(s)
Antimicrobial Cationic Peptides/immunology , Cell Degranulation/immunology , Mast Cells/immunology , Receptors, Lysosphingolipid/immunology , Skin Diseases, Viral/immunology , Skin/immunology , Vaccinia virus/immunology , Vaccinia/immunology , Animals , Antimicrobial Cationic Peptides/genetics , Antimicrobial Cationic Peptides/metabolism , Cell Degranulation/genetics , Immunity, Innate/physiology , Mast Cells/metabolism , Mast Cells/virology , Mice , Mice, Knockout , Receptors, Lysosphingolipid/genetics , Receptors, Lysosphingolipid/metabolism , Skin/metabolism , Skin/virology , Skin Diseases, Viral/genetics , Skin Diseases, Viral/metabolism , Sphingosine-1-Phosphate Receptors , Vaccinia/genetics , Vaccinia/metabolism , Vaccinia virus/metabolism , Virus Inactivation , CathelicidinsABSTRACT
LL-23 is a natural peptide corresponding to the 23 N-terminal amino acid residues of human host defense cathelicidin LL-37. LL-23 demonstrated, compared to LL-37, a conserved ability to induce the chemokine MCP-1 in human peripheral blood mononuclear cells, a lack of ability to suppress induction of the pro-inflammatory cytokine TNF-α in response to bacterial lipopolysaccharides (LPS), and reduced antimicrobial activity. Heteronuclear multidimensional nuclear magnetic resonance (NMR) characterization of LL-23 revealed similar secondary structures and backbone dynamics in three membrane-mimetic micelles: SDS, dodecylphosphocholine (DPC), and dioctanoylphosphatidylglycerol. The NMR structure of LL-23 determined in perdeuterated DPC contained a unique serine that segregated the hydrophobic surface of the amphipathic helix into two domains. To improve our understanding, Ser9 of LL-23was changed to either Ala or Val on the basis of homologous primate cathelicidins. These changes made the hydrophobic surface of LL-23 continuous and enhanced antibacterial activity. While identical helical structures did not explain the altered activities, a reduced rate of hydrogen-deuterium exchange from LL-23 to LL-23A9 to LL-23V9 suggested a deeper penetration of LL-23V9 into the interior of the micelles, which correlated with enhanced activities. Moreover, these LL-23 variants had discrete immunomodulatory activities. Both restored the TNF-α dampening activity to the level of LL-37. Furthermore, LL-23A9, like LL-23, maintained superior protective MCP-1 production, while LL-23V9 was strongly immunosuppressive, preventing baseline MCP-1 induction and substantially reducing LPS-stimulated MCP-1 production. Thus, these LL-23 variants, designed on the basis of a structural hot spot, are promising immune modulators that are easier to synthesize and less toxic to mammalian cells than the parent peptide LL-37.
Subject(s)
Anti-Infective Agents , Cathelicidins/chemistry , Immunologic Factors , Mutation , Peptide Fragments/chemistry , Peptide Fragments/pharmacology , Sequence Homology, Amino Acid , Amino Acid Sequence , Animals , Anti-Infective Agents/chemistry , Anti-Infective Agents/metabolism , Anti-Infective Agents/pharmacology , Chemokines/metabolism , Deuterium Exchange Measurement , Humans , Immunologic Factors/chemistry , Immunologic Factors/genetics , Immunologic Factors/metabolism , Immunologic Factors/pharmacology , Leukocytes, Mononuclear/drug effects , Leukocytes, Mononuclear/immunology , Leukocytes, Mononuclear/metabolism , Membrane Potentials/drug effects , Microbial Sensitivity Tests , Models, Molecular , Molecular Sequence Data , Peptide Fragments/genetics , Peptide Fragments/metabolism , Primates , Protein Conformation , Staphylococcus aureus/drug effectsABSTRACT
A diverse environment challenges skin to maintain temperature, hydration, and electrolyte balance while also maintaining normal immunological function. Rosacea is a common skin disease that manifests unique inflammatory responses to normal environmental stimuli. We hypothesized that abnormal function of innate immune pattern recognition could explain the enhanced sensitivity of patients with rosacea, and observed that the epidermis of patients with rosacea expressed higher amounts of Toll-like receptor 2 (TLR2) than normal patients. Increased expression of TLR2 was not seen in other inflammatory skin disorders such as atopic dermatitis or psoriasis. Overexpression of TLR2 on keratinocytes, treatment with TLR2 ligands, and analysis of TLR2-deficient mice resulted in a calcium-dependent release of kallikrein 5 from keratinocytes, a critical protease involved in the pathogenesis of rosacea. These observations show that abnormal TLR2 function may explain enhanced inflammatory responses to environmental stimuli and can act as a critical element in the pathogenesis of rosacea.
Subject(s)
Keratinocytes/metabolism , Keratinocytes/pathology , Rosacea/metabolism , Serine Proteases/metabolism , Toll-Like Receptor 2/metabolism , Animals , Antimicrobial Cationic Peptides/metabolism , Biopsy , Calcium/metabolism , Cells, Cultured , Disease Models, Animal , Humans , Kallikreins/metabolism , Mice , Mice, Inbred C57BL , Mice, Knockout , Propionibacterium acnes/isolation & purification , Propionibacterium acnes/physiology , Rosacea/pathology , Skin/microbiology , Skin/pathology , CathelicidinsABSTRACT
Statins are inhibitors of 3-hydroxy 3-methylglutaryl coenzyme A (HMG-CoA) reductase, the rate-limiting enzyme in cholesterol biosynthesis. Recent clinico-epidemiologic studies correlate patients receiving statin therapy with having reduced mortality associated with severe bacterial infection. Investigating the effect of statins on the innate immune capacity of phagocytic cells against the human pathogen Staphylococcus aureus, we uncovered a beneficial effect of statins on bacterial clearance by phagocytes, although, paradoxically, both phagocytosis and oxidative burst were inhibited. Probing instead for an extracellular mechanism of killing, we found that statins boosted the production of antibacterial DNA-based extracellular traps (ETs) by human and murine neutrophils and also monocytes/macrophages. The effect of statins to induce phagocyte ETs was linked to sterol pathway inhibition. We conclude that a drug therapy taken chronically by millions alters the functional behavior of phagocytic cells, which could have ramifications for susceptibility and response to bacterial infections in these patients.
Subject(s)
Extracellular Space/microbiology , Hydroxymethylglutaryl-CoA Reductase Inhibitors/pharmacology , Macrophages/drug effects , Neutrophils/drug effects , Pneumonia, Staphylococcal/drug therapy , Staphylococcus aureus/drug effects , Acyl Coenzyme A/antagonists & inhibitors , Animals , Cells, Cultured , DNA, Bacterial/drug effects , DNA, Bacterial/immunology , Extracellular Space/immunology , Extracellular Space/metabolism , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Macrophages/immunology , Macrophages/microbiology , Male , Mice , Mice, Inbred C57BL , Mice, Inbred CFTR , Neutrophils/immunology , Neutrophils/microbiology , Phagocytes/drug effects , Phagocytes/immunology , Phagocytes/microbiology , Pneumonia, Staphylococcal/immunology , Pneumonia, Staphylococcal/microbiologyABSTRACT
Streptococcal inhibitor of complement (SIC) is a highly polymorphic extracellular protein and putative virulence factor secreted by M1 and M57 strains of group A Streptococcus (GAS). The sic gene is highly upregulated in invasive M1T1 GAS isolates following selection of mutations in the covR/S regulatory locus in vivo. Previous work has shown that SIC (allelic form 1.01) binds to and inactivates complement C5b67 and human cathelicidin LL-37. We examined the contribution of SIC to innate immune resistance phenotypes of GAS in the intact organism, using (1) targeted deletion of sic in wild-type and animal-passaged (covS mutant) M1T1 GAS harboring the sic 1.84 allele and (2) heterologous expression of sic in M49 GAS, which does not possess the sic genein its genome. We find that M1T1 SIC production is strongly upregulated upon covS mutation but that the sic gene is not required for generation and selection of covS mutants in vivo. SIC 1.84 bound both human and murine cathelicidins and was necessary and sufficient to promote covS mutant M1T1 GAS resistance to LL-37, growth in human whole blood and virulence in a murine model of systemic infection. Finally, the sic knockout mutant M1T1 GAS strain was deficient in growth in human serum and intracellular macrophage survival. We conclude that SIC contributes to M1T1 GAS immune resistance and virulence phenotypes.
Subject(s)
Bacterial Proteins/metabolism , Cathelicidins/metabolism , Macrophages, Peritoneal/metabolism , Streptococcal Infections/immunology , Streptococcus pyogenes/physiology , Animals , Bacterial Proteins/genetics , Bacteriolysis/genetics , Cathelicidins/immunology , Cell Growth Processes/genetics , Cells, Cultured , Cloning, Molecular , Gene Expression Regulation, Bacterial , Gene Knockout Techniques , Histidine Kinase , Humans , Immune Evasion/genetics , Immunity, Innate , Intracellular Signaling Peptides and Proteins/genetics , Macrophages, Peritoneal/immunology , Macrophages, Peritoneal/microbiology , Macrophages, Peritoneal/pathology , Male , Mice , Mice, Inbred C57BL , Mutation/genetics , Protein Binding/genetics , Species Specificity , Streptococcal Infections/microbiology , Streptococcal Infections/physiopathology , Streptococcus pyogenes/pathogenicity , Transgenes/genetics , Virulence/geneticsABSTRACT
Production of antimicrobial peptides by epithelia is an essential defense against infectious pathogens. In this study we evaluated whether the commensal microorganism Staphylococcus epidermidis may enhance production of antimicrobial peptides by keratinocytes and thus augment skin defense against infection. Exposure of cultured undifferentiated human keratinocytes to a sterile nontoxic small molecule of <10 kDa from S. epidermidis conditioned culture medium (SECM), but not similar preparations from other bacteria, enhanced human beta-defensin 2 (hBD2) and hBD3 mRNA expression and increased the capacity of cell lysates to inhibit the growth of group A Streptococcus (GAS) and S. aureus. Partial gene silencing of hBD3 inhibited this antimicrobial action. This effect was relevant in vivo as administration of SECM to mice decreased susceptibility to infection by GAS. Toll-like receptor 2 (TLR2) was important to this process as a TLR2-neutralizing antibody blocked induction of hBDs 2 and 3, and Tlr2-deficient mice did not show induction of mBD4. Taken together, these findings reveal a potential use for normal commensal bacterium S. epidermidis to activate TLR2 signaling and induce antimicrobial peptide expression, thus enabling the skin to mount an enhanced response to pathogens.
Subject(s)
Bacterial Proteins/metabolism , Staphylococcal Skin Infections/prevention & control , Staphylococcus aureus , Staphylococcus epidermidis , Toll-Like Receptor 2/metabolism , Animals , Antibodies, Neutralizing/immunology , Cells, Cultured , Culture Media, Conditioned/pharmacology , Epidermal Cells , Epidermis/immunology , Epidermis/microbiology , Gene Silencing , Humans , Keratinocytes/cytology , Keratinocytes/microbiology , Mice , Mice, Inbred C57BL , Mice, Knockout , Signal Transduction/immunology , Staphylococcal Skin Infections/immunology , Staphylococcal Skin Infections/metabolism , Streptococcal Infections/immunology , Streptococcal Infections/metabolism , Streptococcal Infections/prevention & control , Streptococcus pyogenes , Toll-Like Receptor 2/immunology , beta-Defensins/genetics , beta-Defensins/metabolismABSTRACT
Antimicrobial peptides play an important role in host defense against pathogens. Recently, phenol-soluble modulins (PSMs) from Staphylococcus epidermidis (S. epidermidis) were shown to interact with lipid membranes, form complexes, and exert antimicrobial activity. Based on the abundance and innocuity of the cutaneous resident S. epidermidis, we hypothesized that their PSMs contribute to host defense. Here we show that S. epidermidis delta-toxin (PSMgamma) is normally present in the epidermis and sparsely in the dermis of human skin using immunohistochemistry. Synthetic delta-toxin interacted with neutrophil extracellular traps (NETs) and colocalized with cathelicidin while also inducing NET formation in human neutrophils. In antimicrobial assays against Group A Streptococcus (GAS), delta-toxin cooperated with CRAMP, hBD2, and hBD3. In whole blood, addition of delta-toxin exerted a bacteriostatic effect on GAS, and in NETs, delta-toxin increased their killing capacity against this pathogen. Coimmunoprecipitation and tryptophan spectroscopy demonstrated direct binding of delta-toxin to host antimicrobial peptides LL-37, CRAMP, hBD2, and hBD3. Finally, in a mouse wound model, GAS survival was reduced (along with Mip-2 cytokine levels) when the wounds were pretreated with delta-toxin. Thus, these data suggest that S. epidermidis-derived delta-toxin cooperates with the host-derived antimicrobial peptides in the innate immune system to reduce survival of an important human bacterial pathogen.
Subject(s)
Anti-Infective Agents/pharmacology , Bacterial Toxins/pharmacology , Peptides/pharmacology , Staphylococcus epidermidis/metabolism , Streptococcus pyogenes/drug effects , Animals , Bacterial Toxins/metabolism , Humans , Immunohistochemistry , Immunoprecipitation , Mice , Mice, Inbred C57BL , Neutrophils/cytology , Protein Binding , Skin/drug effects , Skin/metabolism , Spectrum Analysis/methodsABSTRACT
Antimicrobial peptides serve as a first line of innate immune defense against invading organisms such as bacteria and viruses. In this study, we hypothesized that peptides produced by a normal microbial resident of human skin, Staphylococcus epidermidis, might also act as an antimicrobial shield and contribute to normal defense at the epidermal interface. We show by circular dichroism and tryptophan spectroscopy that phenol-soluble modulins (PSMs) gamma and delta produced by S. epidermidis have an alpha-helical character and a strong lipid membrane interaction similar to mammalian AMPs such as LL-37. Both PSMs directly induced lipid vesicle leakage and exerted selective antimicrobial action against skin pathogens such as Staphylococcus aureus. PSMs functionally cooperated with each other and LL-37 to enhance antimicrobial action. Moreover, PSMs reduced Group A Streptococcus (GAS) but not the survival of S. epidermidis on mouse skin. Thus, these data suggest that the production of PSMgamma and PSMdelta by S. epidermidis can benefit cutaneous immune defense by selectively inhibiting the survival of skin pathogens while maintaining the normal skin microbiome.
Subject(s)
Bacterial Toxins/pharmacology , Keratinocytes/microbiology , Staphylococcus epidermidis/metabolism , Streptococcal Infections/drug therapy , Streptococcus pyogenes , Animals , Antimicrobial Cationic Peptides/metabolism , Bacterial Toxins/chemical synthesis , Cell Membrane Permeability , Cells, Cultured , Circular Dichroism , Epidermal Cells , Epidermis/immunology , Epidermis/microbiology , Humans , Keratinocytes/cytology , Keratinocytes/immunology , Lipid Bilayers/metabolism , Membranes, Artificial , Mice , Mice, Inbred C57BL , Protein Structure, Secondary , Staphylococcal Skin Infections/drug therapy , Staphylococcal Skin Infections/immunology , Staphylococcal Skin Infections/microbiology , Staphylococcus aureus , Streptococcal Infections/immunology , Streptococcal Infections/microbiology , SymbiosisABSTRACT
The normal microflora of the skin includes staphylococcal species that will induce inflammation when present below the dermis but are tolerated on the epidermal surface without initiating inflammation. Here we reveal a previously unknown mechanism by which a product of staphylococci inhibits skin inflammation. This inhibition is mediated by staphylococcal lipoteichoic acid (LTA) and acts selectively on keratinocytes triggered through Toll-like receptor 3(TLR3). We show that TLR3 activation is required for normal inflammation after injury and that keratinocytes require TLR3 to respond to RNA from damaged cells with the release of inflammatory cytokines. Staphylococcal LTA inhibits both inflammatory cytokine release from keratinocytes and inflammation triggered by injury through a TLR2-dependent mechanism. To our knowledge, these findings show for the first time that the skin epithelium requires TLR3 for normal inflammation after wounding and that the microflora can modulate specific cutaneous inflammatory responses.
Subject(s)
Inflammation/microbiology , Skin Diseases, Bacterial/physiopathology , Staphylococcal Infections/physiopathology , Toll-Like Receptor 3/physiology , HumansABSTRACT
LL-37 is a broad-spectrum human antimicrobial peptide in the cathelicidin family. Potency assays in the form of minimal inhibitory concentration and vesicle leakage indicate that the single-tryptophan mutants, F6W and F17W, are as effective at killing bacteria and disrupting membranes as the native, tryptophan-free LL-37 peptide. Steady-state fluorescence and UV resonance Raman spectroscopy of F6W and F17W reveal molecular details of these tryptophan residues. The local environment polarity, hydrogen bond strength of the indole N-H moiety, and rotational freedom decrease for both F6W and F17W in the presence of carbonate ions relative to in pure distilled water; these results are consistent with burial of the hydrophobic region of alpha-helical LL-37 in oligomeric cores induced in the presence of carbonate ions. Differences in the spectroscopic properties of the carbonate-induced alpha-helical forms of F6W and F17W reflect the presence of a local lysine residue near F6W that makes the microenvironment of F6W more polar than that of F17W. In the presence of lipid vesicles, the mutants undergo additional loss of environment polarity, hydrogen bond strength, and rotational freedom. Quenching experiments utilizing brominated lipids reveal that the tryptophan residues in both mutants are essentially equidistant from the bilayer center and that bromines closer to the bilayer center, in the 9,10 positions, quench fluorescence more efficiently than those closer to the headgroups (6,7 positions). These results support carpeting or toroidal pore mechanisms of membrane disruption by LL-37 and demonstrate that the combination of tryptophan mutants and sensitive spectroscopic tools may provide important molecular clues about antimicrobial action.