Human beta defensin levels and vaginal microbiome composition in post-menopausal women diagnosed with lichen sclerosus.

Human beta defensins (hBDs) may play an important role in the progression of lichen sclerosus (LS), due to their ability to induce excessive stimulation of extracellular matrix synthesis and fibroblast activation. The genetic ability of the individual to produce defensins, the presence of microbes influencing defensin production, and the sensitivity of microbes to defensins together regulate the formation of an ever-changing balance between defensin levels and microbiome composition. We investigated the potential differences in postmenopausal vaginal microbiome composition and vaginal hBD levels in LS patients compared to non-LS controls. LS patients exhibited significantly lower levels of hBD1 (p = 0.0003), and significantly higher levels of hBD2 (p = 0.0359) and hBD3 (p = 0.0002), compared to the control group. The microbiome of the LS patients was dominated by possibly harmful bacteria including Lactobacillus iners, Streptococcus anginosus or Gardnerella vaginalis known to initiate direct or indirect damage by increasing defensin level production. Our observations highlight that correcting the composition of the microbiome may be applicable in supplementary LS therapy by targeting the restoration of the beneficial flora that does not increase hBD2-3 production.

Potential role of the skin and gut microbiota in premenarchal vulvar lichen sclerosus: A pilot case-control study.

The etiology of vulvar lichen sclerosus (LS) remains unclear; however, alterations in cutaneous and gut microbiota may be contributing to the pathogenesis of this inflammatory condition. To explore this hypothesis, we conducted a pilot case-control study, obtaining dermal swab and stool samples from prepubertal girls with vulvar LS (n = 5), girls with nonspecific vulvovaginitis (n = 5), and healthy controls (n = 3). Samples (n = 56) were subjected to total DNA extractions. Resulting DNA was purified, subjected to PCR (targeting the V3V4 region of the 16S rRNA gene), sequenced, and analyzed using QIIME, MetagenomeSeq, and DESeq2 software packages. Our findings showed that there were significant differences in the cutaneous and gut microbiotas of girls with LS compared to controls. On the skin, girls with LS had a statistically significantly higher relative abundance of Porphyromonas spp., Parvimonas spp., Peptoniphilus spp., Prevotella spp., Dialister spp., and Peptostreptococcus spp., but a lower relative abundance of Cornyebacterium compared to the control group. In the gut samples, girls with LS had a significantly higher relative abundance of Dialister spp., Clostridiales spp., Paraprevotella spp., Escherichia coli, Bifidobacterium adolescentis, and Akkermansia muciniphila, and a lower relative abundance of Roseburia faecis and Ruminococcus bromii compared to controls. These results suggest a potential association between cutaneous and gut dysbiosis and pediatric vulvar LS. Future studies involving larger samples sizes are warranted to further evaluate this association.

Synchronous genitourinary lichen sclerosus signals a distinct urinary microbiome profile in men with urethral stricture disease.

PURPOSE: Alterations in the urinary microbiome have been associated with urological diseases. The microbiome of patients with urethral stricture disease (USD) remains unknown. Our objective is to examine the microbiome of USD with a focus on inflammatory USD caused by lichen sclerosus (LS). METHODS: We collected mid-stream urine samples from men with LS-USD (cases; n = 22) and non-LS USD (controls; n = 76). DNA extraction, PCR amplification of the V4 hypervariable region of the 16S rRNA gene, and sequencing was done on the samples. Operational taxonomic units (OTUs) were defined using a > 97% sequence similarity threshold. Alpha diversity measurements of diversity, including microbiome richness (number of different OTUs) and evenness (distribution of OTUs) were calculated and compared. Microbiome beta diversity (difference between microbial communities) relationships with cases and controls were also assessed. RESULTS: Fifty specimens (13 cases and 37 controls) produced a 16S rRNA amplicon. Mean sample richness was 25.9 vs. 16.8 (p = 0.076) for LS-USD vs. non-LS USD, respectively. LS-USD had a unique profile of bacteria by taxonomic order including Bacillales, Bacteroidales and Pasteurellales enriched urine. The beta variation of observed bacterial communities was best explained by the richness. CONCLUSIONS: Men with LS-USD may have a unique microbiologic richness, specifically inclusive of Bacillales, Bacteroidales and Pasteurellales enriched urine compared to those with non-LS USD. Further work will be required to elucidate the clinical relevance of these variations in the urinary microbiome.

Presence of Borrelia burgdorferi "Sensu Lato" in patients with morphea from the Amazonic region in Brazil.

BACKGROUND: In the present study, Borrelia spirochetes were found in four (26.6%) out of 15 patients with Atrophoderma of Pasini and Pierini (IAPP) and lichen sclerosis et atrophicans (LSA) from the Brazilian Amazon Region. MATERIAL AND METHODS: Borreliosis was investigated by immunohistochemistry and focus floating microscopy for Borrelia burgdorferi in skin biopsy samples from 15 patients with both clinical and histopathology evidences compatible with Morphea, LSA, and IAPP. RESULTS: Spirochetes were detected by specific immunohistochemistry and focus floating microscopy for B. burgdorferi in samples from three patients. A limitation of our study was the fact that we were not able to isolate and culture these organisms. CONCLUSION: Our data confirm the presence of borreliosis cases in the Amazon.

Borrelia in granuloma annulare, morphea and lichen sclerosus: a PCR-based study and review of the literature.

BACKGROUND: Morphea, granuloma annulare (GA) and lichen sclerosus et atrophicans (LSA) have also been suggested to be linked to Borrelia infection. Previous studies based on serologic data or detection of Borrelia by immunohistochemistry and polymerase chain reaction (PCR) reported contradictory results. Thus, we examined skin biopsies of morphea, GA and LSA by PCR to assess the prevalence of Borrelia DNA in an endemic area and to compare our results with data in the literature. METHODS: Amplification of DNA sequences of Borrelia burgdorferi sensu lato by nested PCR from formalin-fixed and paraffin-embedded skin biopsies of morphea, GA and LSA, followed by automated sequencing of amplification products. PCR-based studies on Borrelia species in these disorders published until July 2009 were retrieved by a literature search. RESULTS: Borrelia DNA was detected in 3 of 112 skin biopsies (2.7%) including one of 49 morphea biopsies (2.0%), one of 48 GA biopsies (2.1%) and one of 15 LSA biopsies (6.6%). Amplification products belonged to B. burgdorferi sensu stricto in two cases available for sequence analysis. CONCLUSIONS: The results of our and most of other PCR-based studies do not argue for a significant association of B. burgdorferi sensu lato with morphea, GA, LSA.

The expanding spectrum of cutaneous borreliosis.

The known spectrum of skin manifestations in cutaneous Lyme disease is continuously expanding and can not be regarded as completed. Besides the classical manifestations of cutaneous borreliosis like erythema (chronicum) migrans, borrelial lymphocytoma and acrodermatitis chronica atrophicans evidence is growing that at least in part also other skin manifestations, especially morphea, lichen sclerosus and cases of cutaneous B-cell lymphoma are causally related to infections with Borrelia. Also granuloma annulare and interstitial granulomatous dermatitis might be partly caused by Borrelia burgdorferi or similar strains. There are also single reports of other skin manifestations to be associated with borrelial infections like cutaneous sarcoidosis, necrobiosis lipoidica and necrobiotic xanthogranuloma. In addition, as the modern chameleon of dermatology, cutaneous borreliosis, especially borrelial lymphocytoma, mimics other skin conditions, as has been shown for erythema annulare centrifugum or lymphocytic infiltration (Jessner Kanof) of the skin.

Infectious balanoposthitis: management, clinical and laboratory features.

BACKGROUND: Balanitis is defined as inflammation of the glans penis, often involving the prepuce (balanoposthitis). It is a common condition due to a wide variety of causes with infection being the most frequent and several microorganisms reported. The clinical aspect is often non specific. The management of balanoposthitis remains a clinical challenge. OBJECTIVE: To evaluate the prevalence of infectious balanitis, its management, clinical features, laboratory procedures and treatment options. SUBJECTS AND METHODS: One hundred eighteen patients with infectious balanitis were evaluated between 1995 and 2004 and laboratory data were collected. RESULTS: Balanitis was diagnosed in 219 (10.7%) of the men that have attended the sexually transmitted disease (STD) Clinic. One hundred eighteen (53.9%) had clinically been assumed to suffer from infectious balanitis. In 75 (63.6%) patients the diagnosis was confirmed by culture studies. Candida albicans was isolated from 24 patients. Staphylococcus spp. and groups B and D Streptococci were the most frequently isolated bacteria. All men were uncircumcised. Ninety-one (77.1%) of infectious balanitis patients were treated with antifungal agents. Twelve patients with infectious noncandida balanitis were treated with general antibiotic therapy. Fifty-five (46.6%) patients had a follow-up of 3 to 12 months during which recurrences were registered in 7 (12.7%) patients. CONCLUSIONS: Infectious balanitis was a common condition, affecting 53.9 % of male STD clinic patients in this study. Candida spp. were the most frequently isolated microorganisms. The clinical aspect is of little value in predicting the infectious agent associated with balanoposthitis.

Possible role of Borrelia burgdorferi sensu lato infection in lichen sclerosus.

OBJECTIVE: To assess the evidence for Borrelia burgdorferi sensu lato infection in patients with lichen sclerosus by focus-floating microscopy. SETTING: Dermatology department of a university hospital. DESIGN: Tissue sections were stained with a polyclonal B burgdorferi antibody using standard histological equipment and then scanned simultaneously in 2 planes: horizontally in a serpentine-like pattern and vertically by focusing through the thickness of the section, ie, focus-floating microscopy. Part of the material was also investigated by Borrelia-specific polymerase chain reaction. PATIENTS: The study population comprised 61 cases of lichen sclerosus and 118 controls (60 negative controls and 68 positive controls). MAIN OUTCOME MEASURE: The presence of B burgdorferi sensu lato within tissue specimens. RESULTS: Using focus-floating microscopy, we detected Borrelia species in 38 of 60 cases (63%) of lichen sclerosus and in 61 of 68 (90%) of positive controls of classic borreliosis, but Borrelia species were absent in all negative controls. Borrelia species were detected significantly more often in early inflammatory-rich (31 of 39 [80%]) than in late inflammatory-poor (7 of 21 [33.3%]) cases (P = .001). Polymerase chain reaction findings were positive in 25 of 68 positive controls (37%) and negative in all 11 cases of lichen sclerosus and all 15 negative controls. CONCLUSIONS: Focus-floating microscopy is a reliable method to detect Borrelia species in tissue sections. The frequent detection of this microorganism, especially in early lichen sclerosus, points to a specific involvement of B burgdorferi or other similar strains in the development or as a trigger of this disease.

IgG antibody reactivity to Borrelia burgdorferi sensu stricto antigens in patients with morphea in Colombia.

BACKGROUND: Morphea and lichen sclerosus et atrophicus (LSA) are sclerotic skin lesions of unknown etiology involving connective tissue. The hypothesis of a borrelial origin of morphea and LSA is currently controversial. METHODS: Immunoglobulin G (IgG) immunoblot serologies against Borrelia burgdorferi in patients with morphea and LSA were analyzed and compared with those from healthy donors and patients with syphilis to determine the association with a probable borrelial agent in Colombia. RESULTS: No significant differences in the number of reactive antigenic bands were found between morphea/LSA patients and syphilis patients or healthy donors. The presence of at least one of the following bands, p28, p39, or p45, was the criterion most able to distinguish morphea/LSA, yielding a specificity of 95% and a sensitivity of 28.6%. Using this criterion as evidence of putative exposure to a causative borrelial agent, sclerotic skin lesions had an odds ratio of 7.60 (95% confidence interval, 1.47-39.23). CONCLUSIONS: These results could be explained by cross-reactivity; however, the partial shared reactivity of sera from patients with syphilis and morphea/LSA does not rule out the possibility that a new spirochetal agent, unrelated to B. burgdorferi or Treponemas, may be the causative agent of morphea/LSA in Colombia.

Isolation and polymerase chain reaction typing of Borrelia afzelii from a skin lesion in a seronegative patient with generalized ulcerating bullous lichen sclerosus et atrophicus.

A 64-year-old woman presented with bullous and ulcerating lichen sclerosus et atrophicus (LSA) on the neck, trunk, genital and perigenital area and the extremities. Histology of lesional skin showed the typical manifestations of LSA; in one of the biopsies spirochaetes were detected by silver staining. Despite treatment with four courses of ceftriaxone with or without methylprednisone for up to 20 days, progression of LSA was only stopped for a maximum of 1 year. Spirochaetes were isolated from skin cultures obtained from enlarging LSA lesions. These spirochaetes were identified as Borrelia afzelii by sodium dodecyl sulphate--polyacrylamide gel electrophoresis and polymerase chain reaction (PCR) analyses. However, serology for B. burgdorferi sensu lato was repeatedly negative. After one further 28-day course of ceftriaxone the lesions stopped expanding and sclerosis of the skin was diminished. At this time cultures for spirochaetes and PCR of lesional skin for B. afzelii DNA remained negative. These findings suggest a pathogenetic role for B. afzelii in the development of LSA and a beneficial effect of appropriate antibiotic treatment.

[Borrelia burgdorferi antibodies in scleroderma circumscripta, lichen sclerosus et atrophicus, erythema nodosum, granuloma annulare, erythema annulare and chronic urticaria]. / Protilátky proti Borrelia burgdorferi pri sclerodermia circumscripta, lichen sclerosus et atrophicus, erythema nodosum, granuloma anulare, erythema anulare a urticaria chronica.

OBJECTIVES: The role of B. burgdorferi in the etiology of sclerodermia circumscripta (SC) and lichen sclerosus et atrophicus (LSA) are is reported in numerous, however controversial studies. The objective of our study is to approximate the solution of the given problem and to widen these consideration by other diagnoses with multifactorial and unclear etiology such as erythema nodosum (EN), granuloma anulare (GA), erythema anulare (EA) and urticaria chronica. MATERIAL AND METHODS: 124 probands were divided into groups according the diagnoses presented above and compared with the negative control group of 131 probands with dermatologic diagnoses, in which the etiologic agent of B. burgdorferi was not assumed and positive group of 55 probands with lyme boreliosis. Indirect immunofluorescent test was used to find out the tieter of antibodies against B. burgdorferi in all groups by using the endemic strains as antigens, which has caused a higher value of the so-called cut-off. RESULTS: The negative control had a positive titer in 44 cases (n = 131, i.e. in 33.6%). The positive titer was found in 11 probands from the SC group (34.4% =, n = 32), 5 probands in LSA (71.4%, n = 7), 9 probands in the EN group (64.3%, n = 14), 6 probands in the EA and GA groups (42.3%, n = 14) and 19 probands in the group of urticaria chronica (33.3%, n = 57). CONCLUSION: We assume that in the probands with high titers of antibodies, B. burgdorferi could play a role in the etiology of the given diseases titers of antibodies against B. burgdorferi. The draft problem could be solved by modern method including PCR with the use of several primers focused on different antigens regarding the certain epidemiologic regions. (Fig. 7, Ref. 17.)

Is morphoea caused by Borrelia burgdorferi? A review.

The aetiology of morphoea and lichen sclerosus et atrophicus is still unknown. Since the detection of Borrelia burgdorferi (B. burgdorferi) as the causative agent of Lyme disease, there has been debate about a possible association between B. burgdorferi and morphoea. Initial serological and cultural studies showed controversial results. The introduction of polymerase chain reaction (PCR) initially suggested an association between B. burgdorferi and morphoea. We reviewed the literature on B. burgdorferi (specific serology, immunohistology, culture, lymphocyte stimulation and DNA detection by PCR) since 1983, using Medline and Current Contents. Histological and immunohistological detection of B. burgdorferi was reported in 0-40% (20 of 82) of the cases with morphoea and in 46-50% (17 of 36) of the cases with lichen sclerosus et atrophicus. Cultivation of spirochetes from lesional skin succeeded in five patients (five of 68) with morphoea, but failed in patients with lichen sclerosus et atrophicus. In Europe and Asia, serological detection of antibodies against B. burgdorferi was described in 0-60% (138 of 609) of patients with morphoea and in 19% (six of 32) in the U.S.A. For lichen sclerosus et atrophicus 0-25% of the published cases (three of 23) in Europe and Asia were seropositive. DNA from B. burgdorferi was detected by PCR in 0-100% (17 of 82) of the tissues of patients with morphoea in Europe and Asia, but not a single case among 98 patients was reported to be positive from the U. S.A. In Europe and Asia, borrelial DNA was detected in 0-100% (nine of 28) of the cases with lichen sclerosus et atrophicus, whereas in the U.S.A. none of 48 patients was positive. There are two possible explanations for these contradictory findings: the most likely is that B. burgdorferi is not a causative agent for morphoea. Another possible explanation could be that a subset of morphoea is caused by a special subspecies of B. burgdorferi that is present in Europe and Asia but does not occur in the U.S.A.

Evidence for Borrelia burgdorferi in morphea and lichen sclerosus.

BACKGROUND: Borrelia burgdorferi (Bb) infection has been implicated in the development of morphea and lichen sclerosus; however, conflicting results have been reported with different investigational methods from different regions. We looked for evidence of Bb in patients with morphea and lichen sclerosus by polymerase chain reaction (PCR) analysis of of skin biopsy samples. METHODS: Formalin-fixed, paraffin-embedded skin biopsy samples from 10 patients with morphea and 12 patients with lichen sclerosus were investigated by PCR analysis for the presence of Bb. RESULTS: The presence of Bb DNA was demonstrated in three of 10 patients with morphea and six of 12 patients with lichen sclerosus by nested PCR. CONCLUSIONS: The data obtained in this study suggest that Bb may play a role in the etiopathogenesis of both morphea and lichen sclerosus at least in the western parts of Turkey.

An infective aetiology for vulval lichen sclerosus re-addressed.

Although there is evidence to support an autoimmune basis for lichen sclerosus, there have also been some studies which suggest an infective aetiology. These include reports of the presence of spirochaetal forms with Steiner silver stains and purplish coccoid forms with Fite stains. We have repeated these studies on vulval biopsies obtained from 16 patients with vulval lichen sclerosus. Using the Steiner silver method we found no evidence of spirochaetal forms in any of the specimens. With the Fite stain we observed purple-staining coccoid forms within the dermis of 13 of the 16 lichen sclerosus specimens. However, these coccoid forms also stained strongly positive with toluidine blue, suggesting they were mast cell granules rather than micro-organisms. We were therefore unable to demonstrate evidence for an infective aetiology in vulval lichen sclerosus, although this cannot yet be excluded. Further work is also needed to understand the significance of mast cells in lichen sclerosus.

No evidence for Borrelia burgdorferi infection in lesions of morphea and lichen sclerosus et atrophicus in Spain. A prospective study and literature review.

The possible association of Borrelia burgdorferi with morphea and lichen sclerosus et atrophicus has been the focus of research and discussion in dermatology during the last 10 years. To investigate the etiopathogenic role of B. burgdorferi in morphea and lichen sclerosus et atrophicus lesions in Spain, we studied 14 cases: 8 patients with lichen sclerosus et atrophicus and 6 with morphea. For the whole group, a prospective study was performed, including serologic studies by indirect immunofluorescence, histologic evaluation of skin biopsy specimens, culture studies, and polymerase chain reaction with different primers sensitive for detecting virtually all B. burgdorferi strains tested to date. Although one patient with morphea had positive serologic findings at low titer, we were not able to culture or detect borrelial DNA in any of the specimens. These findings do not confirm an association between B. burgdorferi and morphea and lichen sclerosus et atrophicus.

Detection of Borrelia burgdorferi DNA (B garinii or B afzelii) in morphea and lichen sclerosus et atrophicus tissues of German and Japanese but not of US patients.

OBJECTIVE: To elucidate the geographic and genospecific association of Borrelia with morphea and lichen sclerosus et atrophicus (LSA). DESIGN: The association of Borrelia burgdorferi with morphea and LSA has been reported, but is still controversial. We conducted a retrospective survey of Borrelia DNA in skin biopsy specimens. SETTINGS: The samples were collected from the outpatient clinic of university hospitals and a dermatopathology laboratory. PATIENTS: Skin biopsy specimens (19 morphea and 34 LSA) were obtained from patients in the United States, Japan, and Germany. DNA samples were subjected to amplification with polymerase chain reaction for B burgdorferi flagellin gene, and for the genotype-specific detection of B burgdorferi sensu stricto, Borrelia garinii, and Borrelia afzelii. RESULTS: Five cases of morphea and 2 cases of LSA in Germany and Japan yielded positive signals for B garinii or B afzelii, the European species. None of the American samples were positive for Borrelia polymerase chain reaction. Borrelia burgdorferi sensu stricto was not detected in any of the specimens. CONCLUSION: Morphea and LSA in Germany and Japan can be related with European genotypes of Borrelia.