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PubMed2007

Biofilm formation by Propionibacterium acnes on biomaterials in vitro and in vivo: impact on diagnosis and treatment.

Propionibacterium acnes is found increasingly as a cause of delayed infection, usually involving implanted biomaterials. Despite susceptibility to common antibiotics, such infections are very difficult to treat and usually require surgical removal of the device. Three clinical isolates of P. acnes were assessed for ability to adhere to titanium, surgical steel and silicone, with and without a plasma conditioning film. After adherence, the biomaterials were then incubated for a further 6 days and examined for biofilm development. All three isolates adhered to all three biomaterials similarly. Importantly, we were able to demonstrate biofilm formation, including production of exopolymer similar in appearance to the polysaccharide intercellular adhesin of Staphylococcus epidermidis. A case summary also demonstrated failure to eradicate P. acnes infection in a hydrocephalus shunt after prolonged treatment. The removed shunt showed obvious biofilm formation, initially obscured by exopolymer when viewed by environmental scanning electron microscopy. Biofilm development by P. acnes explains the difficulties encountered in clinical management of such infections.

Bacterial Adhesion↗
PubMed2001

Role of monocyte chemoattractant protein-1 in Propionibacterium acnes-induced pulmonary granulomatosis.

The inflammatory process in granulomatous disorders such as sarcoidosis is mainly the consequence of delayed hypersensitivity induced by causative antigens. Propionibacterial DNA was isolated recently by PCR from human sarcoidosis tissue. Hence, we developed a model using sensitized rabbits for T cell-mediated pulmonary granulomatosis induced by Propionibacterium acnes (P. acnes) and investigated the role of monocyte chemoattractant protein-1 (MCP-1) in the pathogenesis of the granuloma formation in vivo. Intravenous injection of P. acnes into sensitized rabbits induced massive pulmonary granulomas on day 3. Maximum levels of MCP-1 in sera and bronchoalveolar lavage fluid (BALF) were detected on day 1 and preceded recruitment of monocyte/macrophages and T cells. In BALF, monocyte chemotaxis peaked 1 day after P. acnes challenge, and T cell chemotaxis peaked 3 days after P. acnes challenge. Anti-MCP-1 IgG inhibited monocyte chemotaxis by 80.2% and T cell chemotaxis by 35.7%. Phenotypic analysis of migrating T cells revealed that activated and memory T cells (CD26(+)/CD45RO(+)) but not naive cells were preferentially attracted to BALF. Administration of MCP-1 antiserum in vivo inhibited the development of granulomas in both size 59.9% reduction and number 28.6% reduction, the number of infiltrating leukocytes in BALF, and the expression of adhesion molecules on leukocytes in peripheral blood and BALF. Our data indicate that MCP-1 plays important roles in granuloma formation by attracting and activating specific types of cells in this model. Furthermore, results suggest that the rabbit model resembles human angiocentric granulomatosis and would be useful for investigating the immunopathogenesis of human pulmonary granulomatosis.

Animals↗
PubMed1986

Effects of diethylstilbestrol on Propionibacterium acnes immunomodulation: inhibition of macrophage activation and antitumor activity.

Exposure of mice to diethylstilbestrol (DES) inhibited Propionibacterium acnes-induced antitumor activity in vivo against the B16F10 subcutaneous tumor. The inhibitory effect was associated with inhibition by DES of the characteristic P. acnes induced splenomegaly and changes in splenic and peritoneal macrophages (M phi) cell populations. The characteristic P. acnes induced reductions in M phi alkaline phosphodiesterase I (APD) ectoenzyme activity and in total RNA synthesis, proposed biochemical markers of tumoricidal M phi, were partially or completely reversed in DES-treated mice. As predicted from these in vivo and in vitro results, DES treatment significantly decreased P. acnes activation of M phi antitumor activity in vitro against B16F10 melanoma and Lewis lung carcinoma cells. These data suggest a macrophage activation defect may be involved in the reduced resistance that DES-treated animals exhibit to a variety of neoplastic and microbial challenges.

Animals↗
PubMed1999

Functional roles of MCP-1 in Propionibacterium acnes-induced, T cell-mediated pulmonary granulomatosis in rabbits.

The immunological manifestation of granuloma formations in humans largely depends on the delayed-type hypersensitivity response. We investigated the involvement of monocyte chemoattractant protein-1 (MCP-1) in a rabbit model of T cell-mediated pulmonary granulomatosis. Intravenous injection of Propionibacterium acnes (P. acnes) into sensitized rabbits induced massive and diffuse pulmonary granulomas. Levels of MCP-1 in sera and bronchoalveolar lavage fluids (BALF) peaked before the granuloma formation reached the peak (on days 1 and 3 after challenge, respectively). Chemotactic activities toward monocytes and T cells in BALF were inhibited by anti-MCP-1 IgG by 80 and 36%, respectively. The phenotypic analysis of the migrating T cells revealed that activated and memory T cells rather than naive cells were preferentially attracted to the BALF. Administration of anti-MCP-1 antiserum inhibited the development of granuloma formation in both size and number, the numbers of infiltrating leukocytes in BALF, the expression of adhesion molecules on peripheral monocytes/T cells, and on macrophages/T cells in BALF, and the production of TNF-alpha in the lung. Anti-MCP-1 resulted in a trend toward decreased level of IL-1beta in the lung. The inhibition of the production of these cytokines appeared to be induced indirectly through the inhibition of the recruitment of macrophages that produce these cytokines. The results suggest important roles of MCP-1 in the development of granuloma formation in this model through the attraction and activation of specific types of cells.

Animals↗
PubMed2002

In situ localization of Propionibacterium acnes DNA in lymph nodes from sarcoidosis patients by signal amplification with catalysed reporter deposition.

Sarcoidosis is a systemic granulomatous disease of unknown aetiology. Many genomes of Propionibacterium acnes and P. granulosum have been detected in lymph nodes from patients with sarcoidosis. In situ localization of propionibacterial genomes in sarcoid lymph nodes may help to establish an aetiological link between sarcoidosis and these indigenous bacteria. Formalin-fixed and paraffin-embedded biopsy samples of lymph nodes from nine patients with sarcoidosis, nine patients with tuberculosis, and nine patients with non-specific lymphadenitis as controls were examined by quantitative real-time PCR (QPCR) for P. acnes and by in situ hybridization (ISH) that used catalysed reporter deposition (CARD) for signal amplification with digoxigenin-labelled oligonucleotide probes that complemented 16S rRNA of P. acnes. The signals per 250 micro m(2) of tissue sections were counted from inside and outside the granulomas of sarcoidosis and tuberculosis and from control lymph nodes. The number of genomes by QPCR was examined for correlation with the mean signal count by ISH with CARD. In sarcoid samples, one or several signals were detected in the cytoplasm of some epithelioid cells in granulomas and of many mononuclear cells around granulomas. The mean signal counts were higher (p < 0.001) in granulomatous areas than in other areas of sarcoid lymph nodes. Even in their non-granulomatous areas, counts were higher than in granulomatous areas (p = 0.0023) and non-granulomatous areas (p < 0.001) of tuberculous lymph nodes and control lymph nodes (p = 0.0071). Correlation between the results by QPCR and ISH with CARD was significant (r = 0.86, p < 0.001). The accumulation of P. acnes genomes in and around sarcoid granulomas suggests that this indigenous bacterium may be related to the cause of granulomatous inflammation in sarcoidosis.

Animals↗
PubMed1998

Structural characterization of the entire 1.3S subunit of transcarboxylase from Propionibacterium shermanii.

Transcarboxylase (TC) from Propionibacterium shermanii, a biotin-dependent enzyme, catalyzes the transfer of a carboxyl group from methylmalonyl-CoA to pyruvate in two partial reactions. Within the multisubunit enzyme complex, the 1.3S subunit functions as the carboxyl group carrier. The 1.3S is a 123-amino acid polypeptide (12.6 kDa), to which biotin is covalently attached at Lys 89. We have expressed 1.3S in Escherichia coli with uniform 15N labeling. The backbone structure and dynamics of the protein have been characterized in aqueous solution by three-dimensional heteronuclear nuclear magnetic resonance (NMR) spectroscopy. The secondary structure elements in the protein were identified based on NOE information, secondary chemical shifts, homonuclear 3J(HNHalpha) coupling constants, and amide proton exchange data. The protein contains a predominantly disordered N-terminal half, while the C-terminal half is folded into a compact domain comprising eight beta-strands connected by short loops and turns. The topology of the C-terminal domain is consistent with the fold found in both carboxyl carrier and lipoyl domains, to which this domain has approximately 26-30% sequence similarity.

Amino Acid Sequence↗
PubMed1993

The polyphosphate- and ATP-dependent glucokinase from Propionibacterium shermanii: both activities are catalyzed by the same protein.

The glucokinase (EC 2.7.1.63) from Propionibacterium shermanii phosphorylates glucose using inorganic polyphosphate (poly(P)) or ATP as the phosphate donor. In this investigation, we have purified the glucokinase to homogeneity, using two methods and show that the polyphosphate and ATP-dependent glucokinase activities eluted as a single protein. The protein peak is shown to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, reverse phase HPLC, and N-terminal sequence analysis. The purified protein eluted as a single peak from gel filtration and hydrophobic interaction HPLC columns and was found to display both the poly(P) and ATP glucokinase activities. Likewise, the two activities comigrated on a native isoelectric focusing gel. In addition, two analogues of ATP with different reactive groups displayed different inhibition patterns with respect to ATP and poly(P). The 2',3'-dialdehyde of ATP, whose reactive group is the dialdehyde of the ribose ring, showed competitive and noncompetitive patterns with respect to ATP and poly(P), respectively. While, 5'-p-fluorosulfonylbenzoyl adenosine, whose reactive sulfonyl fluoride group is related to the gamma-phosphoryl group of ATP, displayed competitive inhibition patterns with both ATP and poly(P). These observations provide evidence that the polyphosphate and ATP-dependent glucokinase activities of P. shermanii are the catalytic properties of a single enzyme and that the two substrates may have different binding sites on the enzyme with a common phosphorylating center.

Adenosine Triphosphate↗
PubMed1994

Role of vascular nitric oxide synthase in endotoxin shock of Propionibacterium acnes-sensitized rats.

The sensitivity of animals to endotoxin differs significantly between species. Thus, factors that determine the susceptibility to endotoxin may play important roles in the pathogenesis of septic shock. In order to determine the mechanism responsible for susceptibility to endotoxin, the effect of lipopolysaccharide (LPS) on the circulatory status of Propionibacterium acnes (PA)-sensitized rats was studied. Following the intravenous administration of a low dose of LPS, the arterial blood pressure of PA-treated rats, but not of normal animals, progressively decreased; the PA-sensitized animals died of circulatory shock within 7 h of LPS administration. N omega-nitro-L-arginine (NA) reduced the depressor effect of LPS by an L-arginine-inhibitable mechanism. Administration of LPS markedly increased the level of the inducible type of nitric oxide (NO) synthase in various tissues, including the aorta, of PA-treated rats but not of control animals. LPS also increased plasma levels of nitrate plus nitrite and aortic levels of cGMP. Dexamethasone inhibited the de novo synthesis of NO synthase in the aorta and other tissues and reduced the depressor effect of LPS. These and other findings suggest that induction of nitric oxide synthase in resistant arteries might underlie the pathogenesis of LPS-induced hypotension in PA-sensitized animals and the mechanism responsible for the susceptibility to endotoxin.

Amino Acid Oxidoreductases↗
PubMed1994

Reactions of hydrogen peroxide with superoxide dismutase from Propionibacterium shermanii--an enzyme which is equally active with iron or manganese--are independent of the prosthetic metal.

Propionibacterium shermanii contains a single constitutive superoxide dismutase (SOD) which is active with either iron or manganese incorporated in the same protein moiety. Copper and cobalt can also be incorporated by the bacteria in the active center of the SOD under conditions of metal deficiency, but in this case the enzyme is enzymatically inactive. In contrast to other bacterial SODs, the Fe-SOD of P. shermanii remains highly resistant to inactivation by hydrogen peroxide, as does Mn-SOD. Both SOD types cannot be distinguished by their inactivation patterns. Incubation with hydrogen peroxide results in a concentration- and time-dependent decrease in tryptophan fluorescence, independent of the metal present in the active center. Moreover, the Fe-SOD shows a time-dependent decrease in spin concentration after addition of hydrogen peroxide, which reflects alterations in the environment of the metal rather than a reduction of Fe3+ to Fe2+. No obvious correlations exist, however, between these effects and the enzymatic activity of the enzyme. The resistance of the SODs from P. shermanii to inactivation by hydrogen peroxide seems to be caused by the fact that a tryptophan residue near the metal-chelating histidine-75--which is present in all Fe-SODs being rapidly inactivated by this agent--is exchanged for valine.

Amino Acid Sequence↗
PubMed1997

Effect of Val 73 --> Trp mutation on the reaction of "cambialistic" superoxide dismutase from Propionibacterium shermanii with hydrogen peroxide.

The H2O2 inactivation of the "cambialistic" superoxide dismutases from Propionibacterium shermanii, which is active with either iron or manganese at the active site, has been studied in the native and Val 73 --> Trp mutant enzymes. The wild-type iron-containing form of this enzyme is much more resistant to treatment with H2O2 with respect to the other metal-specific Fe superoxide dismutase isoenzymes. After incubation with high amounts of H2O2 the enzyme maintains more than 40% of the initial activity. The activity of the Val 73 --> Trp mutant drastically decreases to less than 5% of the initial activity after incubation with hydrogen peroxide. Amino acid analysis of the H2O2-treated mutant enzyme evidenced the loss of the Trp 73 residue which is shown to play a critical role in the stabilization of the monomer fold of the enzyme. On the other hand, the manganese-containing wild-type and mutant enzymes were completely resistant toward H2O2 demonstrating the specific role of iron in the inactivation process.

Binding Sites↗
PubMed1995

Metal uptake of recombinant cambialistic superoxide dismutase from Propionibacterium shermanii is affected by growth conditions of host Escherichia coli cells.

We constructed the complete nucleotide sequence coding for the cambialistic superoxide dismutase from Propionibacterium shermanii by ligation of a synthetic linker to a polymerase chain reaction amplification product obtained using degenerate primers. We set up an expression system yielding large amounts of recombinant superoxide dismutase in the cytoplasm of Escherichia coli and purified the enzyme from cells grown in a complex medium. The physicochemical properties of the recombinant enzyme were identical to those of the natural protein. Under anaerobic conditions the enzyme produced in an iron-supplemented medium incorporated iron as metal cofactor, while the enzyme purified from cells grown under aerobic conditions contained a variable amount of iron and manganese depending on metal availability. Functional equivalence of the two metals in this superoxide dismutase variant was indicated by independence of enzyme activity from Fe/Mn ratio.

Amino Acid Sequence↗
PubMed1996

Singlet oxygen (1 delta g) generation from coproporphyrin in Propionibacterium acnes on irradiation.

Although singlet oxygen has been postulated to be a highly reactive and toxic intermediate, there has been no evidence of considerable generation of singlet oxygen in vivo level except for special cases. In this work, we firstly measured the near-infrared emission spectra corresponding to the O2(1 delta g) --> O2(3 epsilon g-) transition of singlet oxygen of cutaneous Propionibacterium acnes (P. acnes) porphyrin under laser excitation. A comparison of the singlet oxygen production of coproporphyrin, which is produced predominantly from P. acnes, with that of other photosensitizers revealed coproporphyrin to be a highly efficient singlet oxygen generator under ultraviolet light A irradiation on the skin. These results suggest that singlet oxygen can be generated on the skin surface from P. acnes porphyrin under ultraviolet irradiation and induce serious damage to the skin.

Coproporphyrins↗
PubMed1994

Propionic acid production by immobilized cells of a propionate-tolerant strain of Propionibacterium acidipropionici.

Cells of the propionate-tolerant strain Propionibacterium acidipropionici P200910, immobilized in calcium alginate beads, were tested for propionic and acetic acid production both in a semidefined laboratory medium and in corn steep liquor in batch, fed-batch, and continuous fermentation. Cell density was about 9.8 x 10(9) cells/g (wet weight) of beads, and beads were added to the medium at 0.1 g (wet weight) beads/ml. Beads could be reused for several consecutive batch fermentations; propionic acid production in the tenth cycle was about 50%-70% of that in the first cycle. In batch culture complete substrate consumption (glucose in semidefined medium, lactate in corn steep liquor) and maximum acid production were seen within 36 h, and acid yields from the substrate were higher than in free-cell fermentations. Fed-batch fermentations were incubated up to 250 h. Maximum propionic acid concentrations obtained were 45.6 g/l in corn steep liquor and 57 g/l in semidefined medium; this is the highest concentration achieved to date in our laboratory. Maximum acetic acid concentrations were 17 g/l and 12 g/l, respectively. In continuous fermentation of semide-fined medium, dilution rates up to 0.31 h-1 could be used, which gave higher volumetric productivities (0.96 g l-1 h-1 for propionic acid and 0.26 g l-1 h-1 for acetic acid) than we have obtained with free cells. Corn steep liquor shows promise as an inexpensive medium for production of both acids by immobilized cells of propionibacteria.

Acetates↗
PubMed1996

Fed-batch fermentation with and without on-line extraction for propionic and acetic acid production by Propionibacterium acidipropionici.

Fed-batch propionic and acetic acid fermentations were performed in semi-defined laboratory medium and in corn steep liquor with Propionibacterium acidipropionici strain P9. On average, over four experiments, 34.5 milligrams propionic acid and 12.8 milligrams acetic acid were obtained in about 146 h in laboratory medium with 79 milligrams glucose added over five feeding periods. The highest concentration of propionic acid, 45 milligrams, was obtained when the glucose concentration was not allowed to drop to zero. In corn steep liquor 35 milligrams propionic acid and 11 milligrams acetic acid were produced in 108 h from 59.4 milligrams total lactic acid provided as seven feedings of corn steep liquor. Extractive fed-batch fermentations were conducted in semi-defined medium using either flat-sheet-supported liquid membranes or hollow-fiber membrane extraction to remove organic acids from the culture medium. As operated during the course of the fermentation, these systems extracted 25% and 22% of the acetic acid and 36.5% and 44.5% of the propionic acid, respectively, produced in the fermentation. Total amounts of acids produced were about the same as in comparable nonextractive fermentations: 30-37 milligrams propionic acid and 13 milligrams acetic acid were produced in 150 h. Limitations on acid production can be attributed to limited substrate feed, not to failure of the extraction system.

Acetic Acid↗
PubMed1982

Immune reaction of tumor-bearing mice to Propionibacterium acnes and the antitumor effect of the bacteria.

The relationship between immunological reaction to Propionibacterium acnes (PA) and the antitumor effect of the injected bacterium was investigated. The aim was to determine whether the strength of the immune reaction to the bacterium can be used to predict its antitumor effectiveness. C3Hf/Sed mice received SC injections (right thigh) of viable cells of a methylcholanthrene-induced fibrosarcoma. When the tumor grew to 5 mm, the hosts received 350 micrograms PA IV as the antitumor treatment. Cellular immunity (footpad test) to PA was assayed in one group of these mice 14 days later, and in the other anti-PA agglutinins were determined 28 days later. The PA injection cured 22 of 58 mice in the first, and 20 of 46 mice in the second group. Footpad reaction and agglutinin titers to PA in cured mice were not statistically different from those in mice eventually killed by the tumor. Therefore, the strength of the immune reaction to PA in tumor-bearing mice could not be used to predict the antitumor effectiveness of the bacterium.

Animals↗
PubMed1983

Immunochemotherapy of breast cancer with Propionibacterium granulosum.

Eighty-two patients with metastatic breast cancer were treated with adriamycin, cyclophosphamide, and 5-fluorouracil (FAC). These patients were divided by randomization into two groups. One was receiving cytostatic drugs only, while another received the identical cytostatic medication plus additional therapy consisting of IV infusions of gamma-radiation-sterilized cells of Propionibacterium granulosum strain KP-45. Metastases were localized in six different sites and/or organs. Multiple parameters for monitoring changes in metastatic lesions and laboratory tests were performed serially in all patients. The addition of P. granulosum to standard (FAC) chemotherapy of metastatic breast cancer influenced metastases in the liver. It also resulted in increased survival time within the observation period. Possible mechanisms of immunomodulating activity of P. granulosum in this disease are discussed.

Adult↗
PubMed1987

The influence of killed Propionibacterium granulosum on experimental infection with Escherichia coli.

Mice infected subcutaneously with 5 X 10(3) viable cells of Escherichia coli ATCC 25922 incorporated in liquified agar developed a systemic infection. Increasing bacterial numbers could be recovered from the liver at several days following infection. Ultimately, the animals died 6 days after infection. Treatment of mice with 1 mg killed Propionibacterium granulosum KP-45 lead to an increased spleen weight at day 7 after intraperitoneal injection. Hence, the animals were highly susceptible to the lethal action of endotoxin. They were, however, markedly protected against infection with E. coli, since definitely lower bacterial counts were found in the liver of pretreated mice in comparison to controls.

Animals↗
PubMed1984

Inhibition of Lewis lung carcinoma in mice by local microwave hyperthermia combined with immunomodulating Propionibacterium granulosum KP-45.

C-57 BL/6 mice with Lewis lung carcinoma were treated 2 weeks after tumor implantation with local microwave hyperthermia (2450 MHz, tumor temperature 43.5 degrees C, 30 min) and/or intratumoral or intraperitoneal injection of 1 mg cell walls of Propionibacterium granulosum KP-45. Tumor growth, survival time of the animals, and the delayed skin hypersensitivity to oxazolone were followed up, as well as the 3H-thymidine uptake of tumorous tissues and the number of lung metastases. The combined treatment of microwave hyperthermia with immunomodulating P. granulosum KP-45 resulted in significantly stronger inhibition of tumor growth than with each of these methods alone. The number of lung metastases could be significantly lowered, and the skin reactivity to oxazolone remained enhanced during the whole observation period (over 70 days after tumor implantation). The implications of the test observation are discussed.

Animals↗