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Cytotoxic activity of Propionibacterium acnes and other skin organisms.

Culture supernatants from four species of skin micro-organisms were tested against VERO (monkey kidney cells) and skin fibroblasts for cytotoxic activity. Cytotoxic activity was produced by the three species of Propionibacterium tested (P. acnes, P. avidum and P. granulosum), and this activity was highest when cultures were grown in the presence of glucose. In contrast, Staphylococcus epidermidis was devoid of cytotoxic activity whether grown in the presence of glucose or not. The agent responsible for the cytotoxic activity was heat stable, of low molecular weight and removable from supernatants by ether extraction. These properties, coupled with the finding that the levels of cytotoxicity are directly proportional to the concentrations of propionate measured in samples, suggests that propionate is the agent responsible for the cytotoxicity of the culture supernatants. Pure propionate and the salts of other carboxylic acids (CI to C5) were tested at the same concentrations and showed different degrees of cytotoxicity depending on their chain length. Propionate may have an important role in the aetiology of the disease acne vulgaris.

Acne Vulgaris↗

Studies of the extracellular proteolytic activity produced by Propionibacterium acnes.

Of several commercial media tested, trypticase soya both containing 0.4% (w/v) D-sorbitol was superior as a growth medium for the production of extracellular proteinase by Propionibacterium acnes (strain P-37). Extracellular proteinase, production of which was shown to be growth-associated by both batch and continuous culture studies, was partially purified by 70% (NH4)2SO4 saturation. Sephadex G-75 chromatography and ion exchange on DEAE-Sephadex A-50. It was shown to be a heterogeneous mixture of at least three molecular species of enzyme. Proteinase I was inhibited by EDTA (10(-3) mol/l) and PMSF (5 millimol/l) and stimulated by CaCl2 (190% at 10(-3) mol/l). It had a molecular weight of 20 to 30000 and a broad pH optimum from 6.5 to 7.5. Proteinase II was an alkaline proteinase with a molecular weight of 30 to 40000 which was not significantly inhibited by EDTA (10(-2) mol/l) nor stimulated by CaCl2. Proteinase III represented a minor proportion of the recovered proteolytic activity, had a molecular weight of 20 to 30000 and was most active in the alkaline pH range. This enzyme was inhibited by EDTA (10(-4) mol/l) and PMSF (5 millimol/l), and stimulated by CaCl2 (250% at 10(-2) mol/l).

Calcium Chloride↗

Comparative in vitro activities of retapamulin (SB-275833) against 141 clinical isolates of Propionibacterium spp., including 117 P. acnes isolates.

Using the NCCLS agar dilution method, we studied the in vitro activity of retapamulin (SB-275833) against 141 clinical isolates of Propionibacterium species, including seven multiresistant strains, and found retapamulin to be the most active agent among those tested with MICs of < or = 1 microg/ml against all isolates.

Anti-Bacterial Agents↗

Investigation of the component of Propionibacterium acnes (Corynebacterium parvum) responsible for macrophage activation.

Mouse peritoneal macrophages, activated with Propionibacterium acnes (Corynebacterium parvum) in vivo, exhibited altered morphological characteristics, increased acid phosphatase activity, and release of [EH]thymidine-labeled DNA from tumor cells in vitro. Comparison with macrophages from mice injected with a control organism, P. jensenii, showed that the morphological changes, but not acid phosphatase, correlated with the development of tumoricidal activity. Investigation of the microbial component responsible for this activity indicated it was heat stable and refractory to extraction by methanol-chloroform, dilute acid, butanol, or a serial combination of extraction procedures. When the cells were disrupted by mechanical breakage, no activity could be found in the cell wall, soluble cytoplasm, or particulate cytoplasm. These results suggest that intact organisms are required for macrophage activation, and they may resolve conflicting reports on the nature of the immunostimulating activity of P. acnes.

Acid Phosphatase↗

Synergistic lysis of erythrocytes by Propionibacterium acnes.

Sheep and human erythrocytes, partially processed by Staphylococcus aureus or Clostridium perfringens, were susceptible to lysis in the presence of Propionibacterium acnes. P. acnes liberated a lipase that was detected on Tween 80 agar and also on phospholipase C-precipitated egg yolk agar. Such a lipase might have contributed in the process of an intensified cellular lysis. Similar reactions were attempted with Lactobacillus acidophilus, known to possess a nondiffusible lipase, and failed to produce any such reactions. The synergistic reactions, between P. acnes and C. perfringens, were compared with The classical CAMP reaction in an attempt to find a correlation with the established membrane composition of the erythrocytes involved. Synergistic reactions observed do seem to reflect the membrane composition. Such findings, besides being contributory to an understanding of the role of these organisms in the process of pathogenesis, are of importance in the elucidation of molecular organization of biomembranes. Detailed studies, involving a large number of representative anaerobic bacteria, may also help provide an avenue in anaerobic species identification.

Anaerobiosis↗

Identification of Propionibacterium acnes and related organisms by precipitin tests with trichloroacetic acid extracts.

The serological identification of Propionibacterium acnes, P. granulosum, and P. avidum, using trichloroacetic acid extracts, is described. With antisera prepared against reference strains, the method has been tested on 142 strains recently isolated from human skin. All except two of the strains could be identified serologically, and there was excellent agreement between the serological results and the fermentation pattern of the strains. Two serological types of P. acnes and two of P. avidum were identified, but only one of P. granulosum.

Antigens, Bacterial↗

Propionibacterium acnes and the pathogenesis of progressive macular hypomelanosis.

BACKGROUND: Progressive macular hypomelanosis is a common hypopigmentation mainly on the central parts of the trunk, predominantly in young adults, especially women. It is often mistaken for pityriasis versicolor and pityriasis alba. It occurs in all races and has been described in many parts of the world. We discovered follicular red fluorescence restricted to lesional skin. We suspected a relation with a porphyrin-producing bacteria residing in sebum of the pilosebaceous duct, and we therefore performed a study in 8 patients. Observation In all biopsy specimens taken from lesional skin of 8 women, we could demonstrate gram-positive bacteria in the pilosebaceous duct, and a mild perifollicular lymphocytic infiltrate was seen. In all but 1 patient, Propionibacterium acnes was yielded from cultured biopsy specimens taken from follicular lesional skin. Healthy follicular skin did not show bacteria in histological sections, and cultures did not yield anaerobic bacteria. CONCLUSIONS: There seems to be a relation between the presence of P acnes and the hypopigmented macules. We propose that a factor is produced by these strains of P acnes, which interfere with melanogenesis. Based on these observations, we are undertaking a clinical trial to find a treatment for this troubling, intractable disease.

Adolescent↗

Chronic postoperative endophthalmitis associated with Propionibacterium acnes.

We present two cases of Propionibacterium acnes endophthalmitis that confirm the association of this organism with a delayed onset and chronic and indolent intraocular inflammation following extracapsular cataract extraction with posterior chamber intraocular lens implantation. The clinical, microbiologic, and histopathologic features of these cases suggest that the organism can be sequestered in the capsular bag, which may make this entity difficult to diagnose. We conclude that complete capsulectomy may be required if inflammation persists after conventional treatment for endophthalmitis, which may include pars plana vitrectomy, partial capsulectomy, and antibiotic therapy. Sterile postoperative uveitis is a diagnosis of exclusion. We stress the importance of appropriate microbiologic investigation in all cases of postsurgical inflammation, even if the onset is delayed until several months after surgery.

Aged↗

Contamination of K-Sol corneal storage medium with Propionibacterium acnes.

K-Sol is a widely used corneal preservation medium. It has an excellent record for corneal storage and no previous report of contamination. During September 1988, on five of our K-Sol-stored donor rims Propionibacterium acnes grew. Cultures of unopened K-Sol also yielded P acnes. This report reviews previous contaminations of corneal storage media and the special characteristics of P acnes that enabled it to survive precautions taken against infection. The detection of P acnes was not obvious, and we emphasize the need for appropriate culturing of all donors, both aerobically and anaerobically, and close scrutiny of delayed inflammation in postoperative patients.

Chondroitin↗

Immunohistochemistry of the inflammatory response in Propionibacterium acnes endophthalmitis.

Specimens were obtained from two patients with culture-proven Propionibacterium acnes endophthalmitis who had undergone vitrectomy. Wright's and Giemsa stains were performed using cytospin preparations of the dilute vitreous and revealed a predominance of polymorphonuclear leukocytes (80% to 90%). The remaining inflammatory cells in the vitreous were mostly macrophages (10% to 15%); very few lymphocytes were present (less than 5%). Immunohistochemical studies using monoclonal antibodies confirmed the paucity of lymphocytes. Most lymphocytes were CD4+ helper/inducer T cells. Almost no CD8+ suppressor/cytotoxic T lymphocytes or B lymphocytes were found. The inflammatory response in these two patients is most characteristic of acute inflammation and consistent with an underlying bacterial infection, despite a clinical picture of persistent, low-grade inflammation. Infection with P acnes has been shown to inhibit CD8+ T cells and may play a role in the persistent inflammation in cases of P acnes endophthalmitis.

Aged↗

Torulopsis candida (Candida famata) endophthalmitis simulating Propionibacterium acnes syndrome.

Four months after undergoing extracapsular cataract extraction with implantation of a posterior chamber intraocular lens, a 74-year-old woman developed granulomatous anterior uveitis. Although she initially responded well to corticosteroid therapy, she experienced multiple recurrences on discontinuation of this therapy. Slit-lamp examination showed the ocular inflammation to be associated with white cortical material within the lens capsular sac. She underwent removal of the implant as well as the lens capsular sac. Anaerobic culture yielded no organisms, but fungus cultures yielded Torulopsis candida. Histopathologic and electron microscopic studies showed large numbers of yeast sequestered within the lens capsular sac and mild granulomatous inflammation around the sac. Torulopsis candida is occasionally isolated from specimens as a contaminant, but has not yet been shown to produce human disease. The case reported herein documents potential pathogenicity of Torulopsis candida and reveals the importance of organisms other than anaerobic bacteria in causing delayed and localized intraocular inflammation that is virtually identical to Propionibacterium acnes infection.

Aged↗

Isolation and characterization of proline iminopeptidase from Propionibacterium freudenreichii ATCC 9614.

A dimeric, 90 kDa subunit intracellular proline iminopeptidase from Propionibacterium freudenreichii ATCC 9614 was purified to homogeneity by chromatography on hydroxyapatite, Sephacryl 200, Phenyl Superose and Mono Q. The enzyme was specific on Pro-p-nitroanilide and Pro-X dipeptides. It hydrolyzed 2 fragments of hormone oligopeptides with an N-terminal proline: bradykinin, f2-7 and substance P, f4-11. A number of oligopeptides containing 5-11 amino acids residues and proline at the penultimate position from N-terminus or other internal position were not hydrolyzed. The enzyme was most active at pH 7-7.5 and at 37-40 degrees C but it retained 9% of maximal activity at pH 5.5 and >12% of maximal activity at 10 or 60 degrees C. The enzyme was inhibited strongly by the serine protease inhibitor 3,4-dichloroisocoumarin, and stimulated markedly by 1 mol/l of NaCl. The results indicate that the enzyme may lead to the accumulation of proline from dipeptides and oligopeptides during the ripening of cheese.

Aminopeptidases↗

Expression and role of CD14 in mice sensitized to lipopolysaccharide by Propionibacterium acnes.

Propionibacterium acnes-primed mice develop an IFN-gamma-dependent hypersensitivity towards LPS. Since CD14 plays a key role in LPS-induced cell activation the regulation and function of CD14 in this sensitization process were studied in IFN-gamma R-/- and the respective wild-type (wt) mice. In unprimed mice, CD14 (mRNA and protein) was either absent (liver) or only weakly expressed in organs (spleen, lung) and in plasma. Priming with P. acnes led to a moderate, mainly IFN-gamma-dependent up-regulation of CD14. LPS challenge of unprimed mice induced an IFN-gamma-independent increase in CD14 mRNA and CD14 protein. LPS challenge of P. acnes-primed mice induced a strong CD14 overexpression. This response was completely absent in IFN-gamma R-/- mice and is therefore strictly IFN-gamma-dependent. The requirement for CD14 in LPS hyper-responsiveness was assessed by comparing CD14-/- and the respective wt mice with respect to their ability to produce TNF and IFN-gamma, two recognized indices of LPS activity. LPS challenge without priming led to a weaker cytokine reaction in CD14-/- than in wt mice. However, priming with P. acnes enhanced the cytokine response to LPS in both wt and CD14-/- mice, although in the latter absolute levels of cytokines were lower. Therefore, hyperreactivity to LPS is characterized by an up-regulation of CD14, but the sensitization by P. acnes is not CD14 dependent.

Animals↗

Propionibacterium acnes isolated from synovial tissue and fluid in a patient with oligoarthritis associated with acne and pustulosis.

This report describes the case of a patient with a 14-month course of severe oligoarthritis associated with acne. Pure cultures of Propionibacterium acnes were isolated from synovial tissue and synovial fluid specimens collected from the same joint after a 4-month interval. After 2 months of treatment with roxithromycin 300 mg/day, rifampicin 1,200 mg/day, and a nonsteroidal antiinflammatory drug (NSAID), followed by 4 months of treatment with azithromycin 1 gm/week and an NSAID, the synovitis persisted. Cultures of skin lesions and synovial fluid at this time were negative. Although P acnes has previously been isolated from bone specimens obtained from patients with osteitis associated with acne, this is the first report of the isolation of this microorganism from the synovial tissue of a patient with arthritis associated with acne. Our findings raise the question of the role of P acnes in the pathogenesis of arthritis associated with acne.

Acne Vulgaris↗

Chronic destructive oligoarthritis associated with Propionibacterium acnes in a female patient with acne vulgaris: septic-reactive arthritis?

Propionibacterium acnes is an anaerobic bacillus implicated in certain chronic arthritides. This report describes an HLA-B27+ 17-year-old woman with acne vulgaris who presented with rapidly destructive arthritis in the left shoulder as well as an evolving left subclavicular adenopathy. One year later, arthritis was detected in the left knee; the inflammatory synovial fluid was sterile. Growth of P acnes was observed in cultures of the shoulder synovium and lymph nodes, but polymerase chain reaction was negative for Borrelia, Chlamydia, and Ureaplasma DNA. Three months of treatment with amoxicillin and rifampicin led to clinical disappearance of the oligoarthritis, but arthritis recurred in the left knee after discontinuation of therapy. On biopsy, bacteria were undetectable in the knee synovium, but chronic arthritis was evident histologically. Antibiotics were reintroduced for 12 months and were again effective against the clinical symptoms. Although the asymmetry, histologic features, arthritis-acne association, and genetic predisposition of this chronic destructive oligoarthritis would seem to indicate a reactive arthropathy, the isolation of P acnes from 2 distinct specimens prompted us to propose calling this a case of septic-reactive arthritis, which is further supported by the absence of progression after antibiotic therapy and the persistence of the rheumatism. To our knowledge, this is the first demonstration of the efficacy of prolonged antibiotic therapy on the joint manifestations of chronic rheumatism associated with acne.

Acne Vulgaris↗

Construction and characterization of ack knock-out mutants of Propionibacterium acidipropionici for enhanced propionic acid fermentation.

Propionibacterium acidipropionici produces propionic acid from glucose with acetic acid, succinic acid, and CO2 as byproducts. In this work, inactivation of ack gene, encoding acetate kinase (AK), by gene disruption and integrational mutagenesis was studied as a method to reduce acetate formation in propionic acid fermentation. The partial ack gene of approximately 750 bp in P. acidipropionici was cloned using a PCR-based method with degenerate primers and sequenced. The deduced amino acid sequence had 88% similarity and 76% identity with the amino acid sequence of AK from Bacillus subtilis. The partial ack gene was used to construct a linear DNA fragment with an inserted tetracycline resistance cassette and a nonreplicative integrational plasmid containing a tetracycline resistance gene cassette. These DNA constructs were then introduced into P. acidipropionici by electroporation, resulting in two mutants, ACK-Tet and TAT-ACK-Tet, respectively. Southern hybridization confirmed that the ack gene in the mutant ACK-Tet was disrupted by the inserted tetracycline resistance gene. As compared to the wild-type, the activities of AK were reduced by 26% and 43% in ACK-Tet and TAT-ACK-Tet mutants, respectively. The specific growth rate of these mutants was reduced by approximately 25% to 0.10/h (0.13/h for the wild-type), probably because of reduced acetate and ATP production. Both mutants produced approximately 14% less acetate from glucose. Although ack disruption alone did not completely eliminate acetate production, the propionate yield was increased by approximately 13%.

Acetate Kinase↗

Dynamics of mixed cultures of Lactobacillus plantarum and Propionibacterium shermanii.

The mixed culture of Lactobacillus plantarum and Propionibacterium shermanii grown anaerobically in glucose minimal medium exhibits features typical of a commensal interaction even though a number of complicating factors, such as a large maintenance requirement of L. plantarum and inhibition of growth of P. shermanii at low pH, are present. A simple mathematical model of the system is presented and is shown to reproduce rather well some of the features of the continuous mixed culture system in both steady-state and transient situations.

Bacteriological Techniques↗

Propionibacterium acnes prosthetic valve endocarditis: a case of severe aortic insufficiency.

Propionibacterium acnes rarely causes systemic disease. Few cases of P. acnes endocarditis have been reported. This report describes a 63-year-old man who presented with severe congestive heart failure. He had prosthetic valve endocarditis which resulted in severe acute aortic insufficiency. During surgery he was found to have complete disruption of the aorta and left ventricle with a false aneurysm encompassing the circumference of the aortic annulus. Cultures of the valve grew P. acnes. Thus, although P. acnes is a rare cause of endocarditis, it may pursue a very aggressive course, especially in the setting of a prosthetic valve.

Aortic Valve↗