Literature DB >> 28386469

Propionibacterium Acnes Phylogenetic Type III is Associated with Progressive Macular Hypomelanosis.

Rolf L W Petersen1, Christian F P Scholz1, Anders Jensen1, Holger Brüggemann1, Hans B Lomholt1.   

Abstract

Progressive macular hypomelanosis (PMH) is a skin disorder that is characterized by hypopigmented macules and usually seen in young adults. The skin microbiota, in particular the bacterium Propionibacterium acnes, is suggested to play a role. Here, we compared the P. acnes population of 24 PMH lesions from eight patients with corresponding nonlesional skin of the patients and matching control samples from eight healthy individuals using an unbiased, culture-independent next-generation sequencing approach. We also compared the P. acnes population before and after treatment with a combination of lymecycline and benzoylperoxide. We found an association of one subtype of P. acnes, type III, with PMH. This type was predominant in all PMH lesions (73.9% of reads in average) but only detected as a minor proportion in matching control samples of healthy individuals (14.2% of reads in average). Strikingly, successful PMH treatment is able to alter the composition of the P. acnes population by substantially diminishing the proportion of P. acnes type III. Our study suggests that P. acnes type III may play a role in the formation of PMH. Furthermore, it sheds light on substantial differences in the P. acnes phylotype distribution between the upper and lower back and abdomen in healthy individuals.

Entities:  

Keywords:  Cutibacterium acnes; Propionibacterium acnes; next-generation sequencing; phylotype; progressive macular hypomelanosis; single locus sequencing type; skin microbiota; subtype III

Year:  2017        PMID： 28386469      PMCID： PMC5372479          DOI： 10.1556/1886.2016.00040

Source DB:  PubMed          Journal:  Eur J Microbiol Immunol (Bp)        ISSN： 2062-509X


Introduction

Progressive macular hypomelanosis (PMH) is characterized by symmetric nonscaly hypopigmented skin areas that are predominantly visible in the sebaceous areas of the trunk. In the lower back and abdomen, discrete lesions are distinguished while they are more confluent on the upper trunk. There is no inflammation, pain, or itching associated with PMH, but the disease can have a major psychosocial effect on patients. PMH appears to be more frequent in young women, and although the disorder has a worldwide distribution, it is most often identified in dark-skinned populations [1-3]. Several treatment modalities are used against PMH including topical benzoylperoxide 5% (BPO) and clindamycin 1% alone or in combination with ultraviolet A (UVA) or narrow band ultraviolet B (UVB) irradiation [4-7], oral lymecycline in combination with topical BPO 5% for 3 months [8], and low-dose isotretinoin for 1 month [9]; however, the ideal treatment is not yet defined. The etiology of PMH is not known; however, several studies indicate that the Gram-positive anaerobic bacterium Propionibacterium acnes may play a pivotal role [10, 11]. Biopsy specimens from PMH lesions contained P. acnes in pilosebaceous ducts in contrast to biopsies from healthy skin [11], real-time PCR showed a significant predominance of P. acnes in lesional skin as compared to nonlesional skin [10], and red fluorescence was detected in lesions when subjected to Woods light [11]. Finally, antibacterial treatment effective against P. acnes leads to repigmentation [4, 8]. It is not known how P. acnes could induce hypopigmentation. Using microscopy, a decrease in melanin production and a change in the distribution of melanosomes with a resultant decrease in epidermal melanin was shown in PMH lesions [12, 13]. Based on multilocus sequence typing (MLST) and single-locus sequence typing (SLST) schemes and complete genome sequencing, the population of P. acnes has been shown to consist of several phylogenetic subtypes commonly designated IA1, IA2, IB, IC, II, and III [14-21]. A previous study based on bacterial cultivation has indicated that certain subtypes of P. acnes may be associated with PMH: an abundance of a specific but unidentified type of P. acnes was observed in PMH lesions that is different from P. acnes types isolated from acne lesions [22]. Recently, we cultured P. acnes isolates belonging to the otherwise uncommon type III from lesions of PMH patients and sequenced their genomes [23], and a very recent study revealed an abundance of P. acnes type III in bacterial cultures from lesional skin in 14 of 34 PMH patients [24]. In the present study, the type distribution of the entire population of P. acnes in affected and unaffected skin areas of PMH patients, including samples after treatment, and matching control samples was determined using a culture-independent next-generation sequencing (NGS)-based SLST approach. Results show a strong association of P. acnes type III with disease.

Materials and methods

Patient and control cohort, treatment regimen

Eight patients with PMH were recruited by voluntary consent in a private dermatology practice in Aalborg, Denmark. The patients were all clinically examined by a specialist in dermatology (H.B. Lomholt), and PMH was diagnosed based on the finding of clinically characteristic lesions, patient history, and a lack of Malassezia in microscopic inspections. All patients were female between 18 and 31 years of age (mean, 23.5 years) (see online). Five of the patients were treated in a 3-month course with oral lymecycline (300 mg, daily) combined with a daily wash using a 5% BPO washing gel, and one patient used only the 5% BPO wash (see online). As controls, eight healthy volunteers were recruited among students at the University of Aarhus, Denmark. They were all female between 24 and 31 years of age (mean, 26.5 years). Information regarding the study participants and treatment is summarized in online.

Sampling sites and procedure

Samples were taken in all patients from three lesional areas including the lower, middle, or upper back (depending on the position of the PMH lesions) and the abdomen, and additional samples were taken from patients in unaffected adjacent skin areas. One patient had no lesions on the abdomen and was sampled from all three positions on the back. Six of the patients were additionally sampled after treatment from the same areas as the pretreatment sample. Samples from the lower and upper back, and abdomen were taken from the eight healthy controls. In addition, samples from the forehead and the buccal cavity were taken from patients (Fig. S1). Samples for NGS-based SLST were obtained by swabbing the skin firmly for 20 s with a sterile cotton swab moistened in sampling buffer (0.1% detergent [Triton X-100] in 0.075 M phosphate buffer, pH 7.9) [25]. DNA was extracted from all collected samples, and SLST fragment amplification was performed as described previously [20]. The amplicons were then subjected to next-generation sequencing (NGS) using the pyrosequencing technology. Subsequent bioinformatics analysis included quality control and sequence read assignment to the STs of the SLST scheme (http://medbac.dk/slst/pacnes), resulting in a high-resolution phylotype analysis of the P. acnes population in each sample. Samples for bacterial cultivation were taken with a sterile charcoal swab moistened in sampling buffer firmly scrubbed on the skin for 20 s. From each patient, two swabs were obtained from lesional skin on the back and abdomen, respectively, and, in addition, two swabs from adjacent nonlesional skin. For a semiquantitative estimation of the number of bacteria, the primary charcoal cotton swabs were streaked on tryptone-yeast-glucose (TYG) agar plates and incubated for 120 h in an anaerobic chamber (Forma Scientific Anaerobic System model 1024).

Next-generation sequencing-based single-locus sequence typing (NGS-based SLST)

Cotton swab samples from both patients and healthy controls were transferred to 1.5-ml Eppendorf tubes, and bacterial DNA was isolated and purified using PowerLyzer® PowerSoil® DNA Isolation Kit (MO BIO, Carlsbad, CA, United States) according to the manufacturer’s instruction. The isolated DNA was then subjected to amplification by PCR as described previously [20]. All primer sequences are given in online. All PCR reactions were made by mixture of 5 μl DNA sample, 2.5 μl AccuPrime PCR Buffer II (Invitrogen), 1.5 μl of forward primer, 1.5 μl of reverse primer (10 μM), 0.15 μl AccuPrime Taq DNA Polymerase High Fidelity (Invitrogen), and 14.35 μl PCR-grade water into a total volume of 25 μl. All samples were amplified by an initial denaturation for 2 min at 94 °C, followed by 35 cycles of 20 s of denaturation at 94 °C, 30 s at 55 °C for annealing, and 60 s of extension at 68 °C, ending with a 5-minute step of extension at 68 °C. Three PCRs per sample were performed and verified on an agarose gel, pooled together, and purified using a NucleoSpin Extract Kit (Macherey-Nagel). The concentration of purified DNA samples was measured with a NanoDrop 2000 spectrophotometer (Thermo Scientific). The amplicons were pooled in batches of 20 and sequenced unidirectionally from the forward primer using Roche GS FLX+ pyrosequencing technology either at Institute of Microbiology and Genetics, Georg-August University Göttingen, Göttingen, Germany or at Eurofins Genomics, Ebersberg, Germany. The data were then processed using the PyroNoise implementation in Mothur v. 1.36.1 [26]. All sequences have been deposited at NCBI with the project number PRJNA347641. The sequence reads were aligned to all the known STs in the SLST database (http://medbac.dk/slst/pacnes) using BLASTn [27] and assigned an individual ST based on a best-hit model with a cut-off value of 99.5%; anything below this threshold or reads with two or more identical best hits were discarded as unassigned reads.

Sanger sequencing-based SLST of bacterial isolates

SLST typing of P. acnes isolates has been described previously [20]. In brief, the sampled charcoal cotton swabs were streaked on tryptone-yeast-glucose (TYG) agar plates and incubated for 72 h in an anaerobic chamber (Forma Scientific Anaerobic System model 1024). This primary culture was then examined; up to 10 random single colonies resembling P. acnes were selected and individually subcultivated on new TYG agar plates for 72 h under anaerobic conditions. After growth, the P. acnes isolates were harvested and subjected to DNA extraction by a boiling procedure at 100 °C for 10 min in 0.5 ml Eppendorf PCR tubes containing 300 μl PCR-grade water. A PCR was carried out with the SLST primers (see online) as follows: 2 μl of the crude DNA extract was mixed with 1 μl of each forward and reverse primer (10 μM), 10 μl 5’-PRIME Hotmastermix (5 PRIME, Hamburg, Germany), and 11 μl of PCR-grade water. PCR conditions were as follows: initial denaturation of 40 s at 96 °C followed by 35 cycles of 35 s of denaturation at 96 °C, 40 s of annealing at 55 °C, and 40 s of extension at 72 °C, followed by a finale 7-minute extension step at 72 °C. The resulting PCR products were run on a 1% agarose gel to ensure quality and were then sequenced at GATC Biotech AG (Konstanz, Germany) with the forward and reverse SLST primers. The sequences were assembled and trimmed to the known SLST fragment size (484 bp) using MEGA v.6.06 [28]. Finally, the resulting sequences were assigned to STs using the SLST database (http://medbac.dk/slst/pacnes).

Statistical analysis

The proportions of P. acnes type III were compared between mean values of the three lesional sites in patients and corresponding sites in controls using the unpaired Wilcoxon rank sum test/Mann–Whitney U test. Lesional sites in patients before and after treatment were compared using the paired Wilcoxon signed rank test.

Ethics statement

The study protocol was approved by the Ethics Committee of Region North, Denmark (document N-20120050), and the study was conducted according to the principles of the declaration of Helsinki. Written informed consent was obtained from all study participants.

Results

Type III P. acnes predominates in PMH samples

SLST amplicons from 96 samples were selected for pyrosequencing. In average, 7911 sequence reads per sample were obtained and 97% of the reads could be assigned to a P. acnes sequence type (ST) (see online). In the whole data set, the SLST scheme distinguished 90 different STs of P. acnes. The results of the P. acnes ST distribution in PMH lesions from eight patients and eight matching healthy controls are shown in ( A clear distinction was found when comparing PMH lesional samples to matching controls: P. acnes type III (the corresponding ST is designated “L”) was the dominating phylotype in PMH lesions (; on average, 73.9% of the reads belonged to this type. In contrast, in the matching control samples, only 14.2% of the reads could be assigned to P. acnes type III (p value 7.8 × 10-[5]). Instead, control samples contained a higher proportion of type IA1 (STs “A” to “E”) strains (56.8%) and type II (ST “K”) strains (22.3%), while these were found in much lower proportions in PMH lesions: type IA1 (12.8%) and type II (5.6%). Individual variation among the patients and controls was observed: the proportion of type III in the total P. acnes population varied between 50% and 91% in PMH lesions and between 0% and 53% in control samples ( We wanted to confirm these findings with a culture-dependent technique: swab samples taken from PMH lesions and healthy controls were cultivated anaerobically. Up to 10 P. acnes colonies per sample were randomly selected from the agar plates; for each isolate, the classical SLST assignment by Sanger sequencing of the PCR-amplified SLST fragment was carried out. In average, 39.6% of the P. acnes colonies were type III in lesion samples, in contrast to only 12.5 % in controls (see online).

P. acnes type III in PMH patients is enriched in PMH regions and adjacent skin areas but is rarely found at other body sites

A recent study [29] suggested that the phylotype distribution of P. acnes is relatively uniform and stable throughout the body. Thus, we wanted to investigate if PMH patients are also predominantly colonized by P. acnes type III at different body sites other than the PMH lesions. Samples from different locations including the lower and upper back, the forehead and the buccal mucosa were analyzed from patients and controls. Type III was rarely detected on the forehead or in the buccal mucosa of patients (. Most patients had a mixture of different P. acnes types on the forehead, in particular, strains of the phylotypes IA2, IC, and II. Only one patient was found to have a significant proportion of type III P. acnes on the forehead. On the buccal mucosa, type IA1 was predominant in most patients. Looking at the P. acnes phylotype distribution of patients on nonlesional skin sites, a dominance of type III strains was detected, albeit at a lesser extent than on lesional skin, 53% versus 74%, respectively ( Healthy individuals were analyzed as well. Interestingly, in average, we could detect a larger proportion of type III strains (21.7%) on the lower back skin compared to the upper back skin (5.0%), indicating that the lower back is the preferred habitat for type III strains ( The dominant P. acnes type on the lower back skin of healthy controls was type II (33.5%), a type that was rarely detected on the upper back (4.0%). The dominating type of the upper back and the abdomen was type IA1 (44.7% and 40.6%, respectively).

PMH treatment alters the P. acnes population and diminishes the proportion of type III strains

Next, we wanted to investigate how the treatment of PMH with a combination of lymecycline and BPO might alter the P. acnes type distribution. Six patients were treated for 3 months, and samples were taken before and after treatment from the same skin location. Information about the treatment regimen and the response for each patient is given in online. Representative images of the back skin of two patients who responded well to the treatment are shown ( We could detect a striking reduction of the proportion of type III P. acnes from an average of 80% before treatment to 22% after treatment (p = 0.015) ( In all six patients, the type III proportion was diminished after treatment in PMH-affected skin sites. In three patients (P2, P4, and P8), the type III population was almost completely eradicated after treatment ( Interestingly, these three patients showed a particular good response to the treatment with almost no remaining PMH lesions. The type distribution after treatment resembled in average the one detected in controls samples ( In contrast, in patients with a less good treatment response, a substantial type III population could be still detected (

Existence of a specific type III linage associated with PMH lesions?

Since type III strains were also detected, albeit fewer, in healthy samples, in particular on the lower back skin, we wanted to investigate if PMH-associated type III strains belong to a specific lineage that is different from health-associated type III strains. Our SLST scheme can differentiate six STs within the type III lineage. The analyses showed that PMH-associated type III P. acnes belong predominantly to the STs L1 (56%) and L6 (25%); the latter ST was detected at lower rates among health-associated type III P. acnes (9%) ( Overall, our data do not reveal a clear-cut difference between the type III populations of healthy and PMH-affected individuals.

Discussion

We report the hitherto most detailed data on the distribution of P. acnes subtypes on multiple skin sites on eight PMH patients and eight healthy controls, using a highly discriminative NGS-based SLST approach. The study revealed an association of one particular type of P. acnes, the phylotype III, with the skin disorder progressive macular hypomelanosis. Moreover, an indication that subtype III is involved in the PMH disease pathogenesis was revealed by the comparison of patient samples before and after treatment: therapy with lymecycline and BPO, both highly active against P. acnes, led to a diminished proportion of type III, which was paralleled by the disappearance of clinical PMH lesions. The applied NGS-based SLST approach gave an average of 7911 reads per sample with 97% of reads assigned to known P. acnes STs. This provided a robust basis for a high-resolution estimate of the type distribution in each sample. Though this technique is regarded as less biased than culture-dependent techniques, a PCR bias due to disproportional amplification of certain sequences cannot be ruled out. Therefore, up to 10 randomly selected colonies were cultured from samples and analyzed by traditional Sanger sequencing for comparison. Importantly, the dominance of type III strains in PMH lesions as compared to controls was consistent in both techniques. P. acnes subtype III was first reported as a new phylogenetic type in 2008 based on four strains isolated from spinal intervertebral disc material [17]. In addition, subtype III isolates were recently detected in surgically excised lumbar disc herniations from 5 of 24 patients [30]. Subtype III bacterial cells differ from other P. acnes types in showing a long filamentous morphology reminiscent of Propionibacterium propionicum. Subtype III differs also in biochemical tests, matrix-assisted laser desorption–ionization time-of-flight mass spectrometry (MALDI-TOF MS) spectra, and genetic markers; thus, it was recently proposed as a new subspecies with the name P. acnes subsp. elongatum [31]. This subtype has rarely been cultured from healthy controls, opportunistic infections, or acne patients [14, 16, 18, 32]. In contrast, the present study showed a striking predominance of subtype III in PMH lesional samples from the lower, middle, and upper back, and abdomen. This corroborates the previous finding of a unique, unidentified P. acnes type in PMH patients [22], and the recent report that type III strains were cultured from lesions of 14 of 34 PMH patients [24]. The NGS-based SLST approach provided no data on the presence of other bacterial species or the total bacterial numbers but only on the proportional distribution of P. acnes subtypes. Previous studies have found a highly significant increase in P. acnes numbers in PMH lesions [11, 24]. In accordance, we could also confirm that P. acnes is more abundant in lesions compared to nonlesional skin: a semiquantitative analysis revealed that all PMH samples from patients had a higher colony-forming unit count as compared to adjacent nonlesional skin (data not shown). As in other studies, no Malassezia fungus was detected and only few bacteria of other species, mainly Staphylococcus epidermidis, were found. Our study revealed a high proportion (74%) of P. acnes type III in lesional skin and also a relatively high proportion (53%) of type III isolates in adjacent nonlesional skin of the patients. In contrast, Barnard et al. cultured P. acnes from nonlesional skin in only one of 34 patient samples, indicative of low P. acnes numbers in nonlesional skin. Therefore, the type distribution at normal skin sites could not be assessed in their study [24]. In addition, the difference may reflect different sampling and sample processing approaches in the two studies. Barnard et al. cultivated bacteria from a homogenized 4-mm punch skin biopsy sample, whereas we used surface skin swabs from a circular area of approximately 1.5 cm in diameter. We noticed that it is difficult to completely separate lesions from adjacent nonlesional skin sites as lesions may be small and plenty and not well defined. This may explain the relatively high proportion of type III strains in nonlesional skin of patients in the present study. Normal controls harbored only a minor proportion of P. acnes type III on corresponding skin sites. The SLST scheme can distinguish six different STs among type III strains and four of these were detected in PMH lesions with no clear differences in their relative proportions in patients and controls. Interestingly, a regional difference in prevalent type III clones was suggested, based on comparisons of PMH isolates from Europe and Brazil [24]. The mechanism leading to macular hypomelanosis is not known, but ultrastructural studies have shown less melanized and aggregated melanosomes instead of single mature melanosomes transferred from melanocytes to keratinocytes [12]. The association with P. acnes type III suggests that a type III-specific factor could be involved. Comprehensive comparison of type III genomes to P. acnes genomes of other subtypes has identified several genomic regions specific to type III genomes encoding functions such as type II secretion system, ABC transporter, inositol transport/modification, gyrase, integrase, transposase, oligopeptide transport, and processing of sugars/amino acids [24, 33, 34]. In addition, some genes are absent from type III genomes but present in all other types including hyaluronate lyase, magnesium-chelatase, iron transporter, bacteriocin, 3-isopropylmalate dehydrogenase, maltose transporter, and periplasmic binding protein. It has to be 2investigated in the future if and which factors of P. acnes type III are important in PMH pathogenesis. At present, there is a major interest in defining the normal human skin microbiome as it is considered important for maintaining healthy skin. Several metagenomic studies have described the skin microbiome on species level, but it has become increasingly clear that subtypes within species may play important roles. Most previous P. acnes studies have shown a predominance of type IA1 followed by type II and IB among skin isolates derived from the face or upper back [14, 16, 18]. The present study is the most detailed study on P. acnes subtype distribution on different body sites. It was shown that subtype III is not normally dominant in skin areas of healthy skin; it was rarely found on the face and in the oral cavity. On healthy skin, type III is more frequent at the lower back and abdomen, together with type II, relative to other body sites. The present study has some limitations. The number of patients and controls is low, even though many samples were analyzed from each person. In general, nonlesional skin was sampled lateral to the lesional area and, therefore, not on exactly corresponding skin areas. After treatment, faint residual lesions in three patients were detectable, even though the clinical response was satisfactory. We do not know if the bacteria most important in the disease process reside on the skin or in the follicles and the surface swab technique employed may have missed bacteria residing in follicles. Furthermore, detailed data on the total number of bacteria at each site and the presence and proportions of other microorganisms besides P. acnes were not obtained in this study. In conclusion, the present study showed that P. acnes phylotype III is associated with PMH lesions and, therefore, may play a role in the disease pathogenesis. In the normal human microbiome, P. acnes subtype III appears to constitute only a minor portion, mainly residing on the lower back and abdomen, among the P. acnes population. The findings open for future studies of specific type III traits to further define the role of the bacterium and increase our understanding of the PMH disease pathogenesis.
Table S1.

Data on eight patients (P) and eight controls (C) included in the study

SexAgeSexAge
P1Female19C1Female28
P2Female24C2Female31
P3Female18C3Female27
P4Female22C4Female26
P5Female24C5Female25
P6Female31C6Female25
P7Female29C7Female26
P8Female21C8Female24
Table S2.

Treatment and treatment responses in six PMH patients treated with benzoylperoxide (BPO) daily washes and/or peroral lymecycline (LC) 300 mg once daily for three months

Patient no.TreatmentOutcome
P1BPO, LCGood response with a few residual lesions
P2BPO, LCGood response
P3Irregular BPO, no LCImproved with residual lesions
P4BPO, LCGood response
P7Irregular BPO, irregular LCGood response with a few residual lesions
P8BPO, LCGood response
Table S3.

Primers used in this study

Primers for NGS-based SLST
SLST_PA_REV5’-CCTATCCCCTGTGTGCCTTGGCAGTCTCAG-CCGGCTGGCAAATGAGGCAT-3’
SLST_PA_MID15’-CCATCTCATCCCTGCGTGTCTCCGACTCAG-ACGAGTGCGT-CAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID25’-CCATCTCATCCCTGCGTGTCTCCGACTCAG-ACGCTCGACA-CAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID35’-CCATCTCATCCCTGCGTGTCTCCGACTCAGAGACGCACTCCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID45’-CCATCTCATCCCTGCGTGTCTCCGACTCAGAGCACTGTAGCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID55’-CCATCTCATCCCTGCGTGTCTCCGACTCAGATCAGACACGCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID65’-CCATCTCATCCCTGCGTGTCTCCGACTCAGATATCGCGAGCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID75’-CCATCTCATCCCTGCGTGTCTCCGACTCAGCGTGTCTCTACAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID85’-CCATCTCATCCCTGCGTGTCTCCGACTCAGCTCGCGTGTCCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID105’-CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTCTATGCGCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID115’-CCATCTCATCCCTGCGTGTCTCCGACTCAGTGATACGTCTCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID135’-CCATCTCATCCCTGCGTGTCTCCGACTCAGCATAGTAGTGCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID145’-CCATCTCATCCCTGCGTGTCTCCGACTCAGCGAGAGATACCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID155’-CCATCTCATCCCTGCGTGTCTCCGACTCAGATACGACGTACAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID165’-CCATCTCATCCCTGCGTGTCTCCGACTCAGTCACGTACTACAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID175’-CCATCTCATCCCTGCGTGTCTCCGACTCAGCGTCTAGTACCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID185’-CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTACGTAGCCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID195’-CCATCTCATCCCTGCGTGTCTCCGACTCAGTGTACTACTCCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID205’-CCATCTCATCCCTGCGTGTCTCCGACTCAGACGACTACAGCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID215’-CCATCTCATCCCTGCGTGTCTCCGACTCAGCGTAGACTAGCAGCGGCGCTGCTAAGAACTT-3
SLST_PA_MID225’-CCATCTCATCCCTGCGTGTCTCCGACTCAGTACGAGTATGCAGCGGCGCTGCTAAGAACTT-3
Primers for Sanger-sequencing-based SLST
Sanger_SLST_for5’-CGCCATCAAGGCACCAACAA-3’
Sanger_SLST_rev5’-ATATCGGCCCGTATTTGGGC-3’
Table S4.

Next-generation sequencing data: numbers of sequence reads and ST assignment in 96 samples

PatientP1P1P1P1P1P1P1P1P1P1P1P2P2P2P2P2P2P2P2P2P3P3P3P3
Sampling spotLBLBUBUBABDFM2-LB2-UB2-UB2-ABDLBLBMBABDF2-LB2-MB2-ABD2-ALBLBUBUB
Number of reads6661109513840533610036148837813938505969063794974438397583189772837603046248518510144744020645196
Number of assigned reads634110762317552509992147157603872500268562454928433096973127764136652972245018410023731519995121
% unassigned reads4.801.7317.321.610.441.132.691.681.130.722.100.921.210.631.941.132.532.431.410.541.191.683.151.44
Unassigned32018966586441682166575134465361628795743511211256575
A112194261198094611233841412398167498618469615351435120041321385127737
A20000022001000000000000000
A3000000000000000000000100
A4000000000000000000000100
A5000000101000100000000006
A60000000010202011321902305
A70004061020000020006000000
A8000002001000000001000100
A9000000000020000000000100
A10000000000000000000000000
A11000000001000000050000000
A12000000000000000000000000
A13000100000010000000000000
A14000000000000000000000000
A15000000000000000000000300
A16000100000010000000000000
A17000002000000000001001001
A18020300101090104000000800
A19000000210020000011001000
A20000000200030002010000001
A21000000004020000000000000
A22000000201010000000000000
A230000000015020002001000200
B10210300000001100001800156120102
C1000014761210306011071726004701137
C2000000000000000000000201
C3000000000000000000000100
C4000000000000000010000100
D100000112040120648134831132906328991504
D2000000000000000000000000
D3000000000000000000000001
E1000000000000000000000000
E2000000000000000000000000
E3000001000000000000000002
E4000000000000000000000000
E5000000000000000000000000
E6000000000000000000000020
E7000000000000001001101021804122
E8000000000000000000000000
E90000000000003030000170503159883
F10304122525201015100020000000000
F2000000000000000000000000
F3000005000010000000000000
F400000674900010020013441025463216278
F5000000000000000000000100
F6000000000000000000000100
F7080122168973383303650000000000000
F80000011800000000000000111
F9000000000000000000000000
F10000000000000000000000000
G10000000000002675001691408400001
H1000220415811821010161315590718633518001114
H2000005000000000010100000
H3000000000000000000000000
H40000000000001400000000000
H5000000000000000010000000
K1377383002000044106042566235544003861141187139
K2305722010020141702812148742117769000
K30019000000000000300000000
K4000000630026000000000000017
K5000000000000000400000000
K6000000000000000200000000
K7000200000140010004700010100
K8800300620275854317121450347261011719490623442109167570900
K9000000000000000000100000
K10000000000000000000000000
K11000000000000000000000000
K12000000000000002000000000
K13000100000000000000000000
K14000000000000000000000000
L15288103752191200298227331143131770271595035705230000253
1214938145136560035879038021140207000010
L30014000000000000000000000
L4000000000000000000000000
L52527164001000490765002100000121
L60101411700051420000009623383913492245
PatientP3P3P3P3P3P4P4P4P4P4P4P4P4P4-2P5P5P5P5P5P6P6P6P6P6
Sampling spotABDF2-LB2-UB2-ABDLBLBMBUBF2-LB2-UB2-UB2-UB-2LBLBABDCHFLBLBUBABDF
Number of reads119897710118311624014294114535424161439719593343813373448781076681858127086808142546239422574041352812788
Number of assigned reads118167493114431446713916112735296157029234585842883316441880356431816125276613131766111415871221324512196
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  34 in total

1.  Comparison of the clinical efficacy of NBUVB and NBUVB with benzoyl peroxide/clindamycin in progressive macular hypomelanosis.

Authors:  J H Sim; D J Lee; J S Lee; Y C Kim
Journal:  J Eur Acad Dermatol Venereol       Date:  2011-02-23       Impact factor: 6.166

2.  Dissecting the taxonomic heterogeneity within Propionibacterium acnes: proposal for Propionibacterium acnes subsp. acnes subsp. nov. and Propionibacterium acnes subsp. elongatum subsp. nov.

Authors:  Itaru Dekio; Renata Culak; Raju Misra; Tom Gaulton; Min Fang; Mitsuo Sakamoto; Moriya Ohkuma; Kenshiro Oshima; Masahira Hattori; Hans-Peter Klenk; Dunstan Rajendram; Saheer E Gharbia; Haroun N Shah
Journal:  Int J Syst Evol Microbiol       Date:  2015-09-30       Impact factor: 2.747

3.  [Progressive and confluent hypomelanosis of the melanodermic metis].

Authors:  G Guillet; R Helenon; M H Guillet; Y Gauthier; N Ménard
Journal:  Ann Dermatol Venereol       Date:  1992       Impact factor: 0.777

4.  Clinical, pathologic, and ultrastructural studies of progressive macular hypomelanosis.

Authors:  Xin-gang Wu; Ai-e Xu; Xiu-zu Song; Jun-hui Zheng; Ping Wang; Hong Shen
Journal:  Int J Dermatol       Date:  2010-10       Impact factor: 2.736

5.  Clonality and anatomic distribution on the skin of antibiotic resistant and sensitive Propionibacterium acnes.

Authors:  Hans B Lomholt; Mogens Kilian
Journal:  Acta Derm Venereol       Date:  2014-09       Impact factor: 4.437

6.  Propionibacterium acnes and the pathogenesis of progressive macular hypomelanosis.

Authors:  Wiete Westerhof; Germaine N Relyveld; Melanie M Kingswijk; Peter de Man; Henk E Menke
Journal:  Arch Dermatol       Date:  2004-02

7.  Ultrastructural findings in progressive macular hypomelanosis indicate decreased melanin production.

Authors:  G N Relyveld; K P Dingemans; H E Menke; J D Bos; W Westerhof
Journal:  J Eur Acad Dermatol Venereol       Date:  2008-02-04       Impact factor: 6.166

8.  Population genetic analysis of Propionibacterium acnes identifies a subpopulation and epidemic clones associated with acne.

Authors:  Hans B Lomholt; Mogens Kilian
Journal:  PLoS One       Date:  2010-08-19       Impact factor: 3.240

9.  Pan-genome and comparative genome analyses of propionibacterium acnes reveal its genomic diversity in the healthy and diseased human skin microbiome.

Authors:  Shuta Tomida; Lin Nguyen; Bor-Han Chiu; Jared Liu; Erica Sodergren; George M Weinstock; Huiying Li
Journal:  mBio       Date:  2013-04-30       Impact factor: 7.867

10.  Genotypic and antimicrobial characterisation of Propionibacterium acnes isolates from surgically excised lumbar disc herniations.

Authors:  Jess Rollason; Andrew McDowell; Hanne B Albert; Emma Barnard; Tony Worthington; Anthony C Hilton; Ann Vernallis; Sheila Patrick; Tom Elliott; Peter Lambert
Journal:  Biomed Res Int       Date:  2013-08-28       Impact factor: 3.411
View more
  12 in total

Review 1.  Acne, the Skin Microbiome, and Antibiotic Treatment.

Authors:  Haoxiang Xu; Huiying Li
Journal:  Am J Clin Dermatol       Date:  2019-06       Impact factor: 7.403

2.  Correlation between hemolytic profile and phylotype of Cutibacterium acnes (formerly Propionibacterium acnes) and orthopedic implant infection.

Authors:  Julia Lee; Kerryl E Greenwood Quaintance; Audrey N Schuetz; Dave R Shukla; Robert H Cofield; John W Sperling; Robin Patel; Joaquin Sanchez-Sotelo
Journal:  Shoulder Elbow       Date:  2019-08-09

3.  Prevalence of Flp Pili-Encoding Plasmids in Cutibacterium acnes Isolates Obtained from Prostatic Tissue.

Authors:  Sabina Davidsson; Jessica Carlsson; Paula Mölling; Natyra Gashi; Ove Andrén; Swen-Olof Andersson; Elzbieta Brzuszkiewicz; Anja Poehlein; Munir A Al-Zeer; Volker Brinkmann; Carsten Scavenius; Seven Nazipi; Bo Söderquist; Holger Brüggemann
Journal:  Front Microbiol       Date:  2017-11-16       Impact factor: 5.640

Review 4.  Over a Decade of recA and tly Gene Sequence Typing of the Skin Bacterium Propionibacterium acnes: What Have We Learnt?

Authors:  Andrew McDowell
Journal:  Microorganisms       Date:  2017-12-21

5.  Draft Genome Sequence of Propionibacterium acnes subsp. elongatum Strain Asn12.

Authors:  Andrew McDowell; Judit Hunyadkürti; Márta Magyari; Andrea Vörös; Balázs Horváth; Sheila Patrick; István Nagy
Journal:  Microbiol Resour Announc       Date:  2018-07-19

6.  Common skin bacteria protect their host from oxidative stress through secreted antioxidant RoxP.

Authors:  Tilde Andersson; Gizem Ertürk Bergdahl; Karim Saleh; Helga Magnúsdóttir; Kristian Stødkilde; Christian Brix Folsted Andersen; Katarina Lundqvist; Anders Jensen; Holger Brüggemann; Rolf Lood
Journal:  Sci Rep       Date:  2019-03-05       Impact factor: 4.379

Review 7.  Potential Role of the Microbiome in Acne: A Comprehensive Review.

Authors:  Young Bok Lee; Eun Jung Byun; Hei Sung Kim
Journal:  J Clin Med       Date:  2019-07-07       Impact factor: 4.241

8.  The Skin Bacterium Propionibacterium acnes Employs Two Variants of Hyaluronate Lyase with Distinct Properties.

Authors:  Seven Nazipi; Kristian Stødkilde-Jørgensen; Carsten Scavenius; Holger Brüggemann
Journal:  Microorganisms       Date:  2017-09-12

Review 9.  A Janus-Faced Bacterium: Host-Beneficial and -Detrimental Roles of Cutibacterium acnes.

Authors:  Holger Brüggemann; Llanos Salar-Vidal; Harald P M Gollnick; Rolf Lood
Journal:  Front Microbiol       Date:  2021-05-31       Impact factor: 5.640

10.  Porphyrin Production and Regulation in Cutaneous Propionibacteria.

Authors:  Emma Barnard; Tremylla Johnson; Tracy Ngo; Uma Arora; Gunilla Leuterio; Andrew McDowell; Huiying Li
Journal:  mSphere       Date:  2020-01-15       Impact factor: 4.389
View more

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