Anti-phage serum antibody responses and the outcome of phage therapy.

We examined the appearance of serum anti-phage antibodies in 25 patients with chronic sinusitis treated with phage therapy (PT). Approximately 30% of patients with weak antibody responses responded positively to PT, which was similar to the results of treatment achieved in a group of patients with high antibody production. In addition, there was no correlation between antibody level and the outcome of PT. These data, derived from a homogenous group of patients, confirm our earlier findings suggesting that the prognostic significance of serum anti-phage antibodies for the outcome of PT should be determined by relevant clinical trials.

Therapeutic potential of phages in autoimmune liver diseases.

Autoimmune liver disease (ALD) poses a difficult medical challenge, as there is a significant number of patients in whom current therapy offers questionable or no benefit, yet its side effects may be serious, including the development of malignancy. Bacterial viruses (phages) have been recognized increasingly as immunomodulators contributing to immune homeostasis and curbing inflammation. Accumulating data suggest that phages may be useful in immunotherapy of ALD. Phages have been shown to down-regulate the expression and/or production and activity of factors associated with hepatic injury [reactive oxygen species, Toll-like receptor (TLR)-4 activation, nuclear factor kappa B (NF-κB) activation, proinflammatory and procoagulant activities of platelets] and up-regulate the expression and/or production of factors demonstrated as playing a protective role [interleukin (IL)-10, IL-1 receptor antagonist].

The effect of pregnancy on humoral rejection in patients after vascularized organ transplantation.

The aim of the study was to evaluate the effect of pregnancy on the production of donor- and nondonor-specific anti-human leukocyte antigen antibodies (anti-HLA Abs) in organ allograft recipients. The study group included four pregnant kidney (RT) and four liver (LT) transplant recipients. The genotype of HLA class I (A, B) and class II (DR) antigens was assessed. Anti-HLA antibodies class I and II were evaluated between 36 and 40 weeks' gestation. Two different control groups consisted of the following: group I (n=8) with nonpregnant RT (n=6) and LT recipients (n=2), and group II with healthy pregnant women (n=10) with anti-HLA Abs detected between 38 and 41 weeks' gestation. The HLA genotype was determined in fathers of the fetuses from the study group and group II controls. Half of group II controls had donor-specific anti-HLA (A, B, and/or DR) Abs, while nondonor-specific anti-HLA Abs were detected in all subjects from that group. Anti-HLA Abs were found in all group II controls. In the study group, anti-HLA Abs were found in only two LT recipients and one RT recipient, but they were not confirmed as donor-specific. Anti-HLA antibodies were not detected in the study group, whereas six out of ten group II controls had anti-HLA Abs against the HLA of the child's father. Pregnancy in vascularized organ recipients does not trigger the mechanism of humoral rejection involving anti-HLA class I and II antibodies with a potentially adverse impact on graft function.

Application of microbiological quantitative methods for evaluation of changes in the amount of bacteria in patients with wounds and purulent fistulas subjected to phage therapy and for assessment of phage preparation effectiveness (in vitro studies).

PURPOSE: Quantitative microbiological studies may provide important information required for successful phage therapy (PT), however methods for PT monitoring of purulent wounds and fistulas has never been reported before. Therefore our goal was to determine and apply microbiological quantitative methods (MQMs) for monitoring experimental PT. METHODS: Samples from agar plates with growing bacteria were collected using dry and wet sterile compresses, or swabs. After shaking the sample in saline the amount of bacteria in suspension was determined. The method was standardized. The MQM using compress was applied for comparison of in vitro activity of phage preparations with other agents for wound rinsing. The usefulness of this swabbing method was tested in the Phage Therapy Unit for monitoring of experimental PT of patients with chronic wounds or purulent fistulas. RESULTS: Minimum, maximum and standard deviation values used for standardization of the studied method showed that data repeatability was good; thus the method was used for quantitation of bacteria taken both from plates in vitro and patients samples. Effectiveness of phage preparations was compared to gentamicin in vitro. Phages were as effective as antibiotics in reducing the amount of bacteria on agar plates, and this effect was not only due to simple mechanical removal of bacteria, but dependent on their antibacterial activity. We have also observed that the results of bacteria quantitation may correlate with the local status of a wound/fistula in a particular stage of PT. CONCLUSION: The standardized swabbing method of bacteria quantitation can be used for PT monitoring. Presented MQMs are simple and may help to monitor the therapy process and to decide on its duration, frequency and a kind of the phage applied. They can also be applied in other antibacterial treatment strategies.

The influence of external factors on bacteriophages--review.

The ability of bacteriophages to survive under unfavorable conditions is highly diversified. We summarize the influence of different external physical and chemical factors, such as temperature, acidity, and ions, on phage persistence. The relationships between a phage's morphology and its survival abilities suggested by some authors are also discussed. A better understanding of the complex problem of phage sensitivity to external factors may be useful not only for those interested in pharmaceutical and agricultural applications of bacteriophages, but also for others working with phages.

Eradication of Enterococcus faecalis by phage therapy in chronic bacterial prostatitis--case report.

The treatment of three patients suffering from chronic bacterial prostatitis who were qualified for an experimental phage therapy protocol managed at the Phage Therapy Unit in Wroclaw is described. They had previously been treated unsuccessfully with long-term targeted antibiotics, autovaccines, and laser biostimulation. Rectal application of phage lysates targeted against Enterococcus faecalis cultured from the prostatic fluid gave encouraging results regarding bacterial eradication, abatement of clinical symptoms of prostatitis, and lack of early disease recurrence.

[VEGF, TNF-alpha and IL-6 in the peritoneal fluid of women with unexplained infertility]. / VEGF, TNF-alpha i IL-6 w plynie otrzewnowym kobiet z nieplodnoscia niejasnego pochodzenia.

Unexplained infertility is an important problem in diagnosis and therapy in everyday gynecologist's practice. Looking for possible reasons of the unexplained infertility we studied the concentrations of selected cytokines (VEGF, TNF-alpha and IL-6) in the peritoneal fluid of women suffering from the unexplained infertility. We compared the results in the studied group with the control group and with the patients with endometriosis. Immunological disorders of the peritoneal fluid in endometriosis are thought to take part in its pathomechanism. Our results suggest that the levels of one of the main factors of endothelium proliferation (VEGF) in the peritoneal fluid from women with unexplained infertility and women with endometriosis are comparable. Concentration IL-6 and TNF-alpha in the peritoneal fluid in case of unexplained infertility and control group was lower than in the endometriosis patients.

Novel nociceptin analogues.

A series of new nociceptin analogues containing cysteine was tested for their nociceptive effects in tail-flick test on mice after icv injection. The cysteines were introduced in order to get irreversibly binding analogues based on the assumption that the cysteines in the ligand can interact with the cysteines from the receptor to form an S-S bridge. In vivo tests revealed that Cys1-nociceptin (1-13)-NH2 (Cys1-NC) is an antagonist, whereas Cys7-NC is an agonist. Gly1[Phe(p-NO2)]4-NC was less active indicating that the antagonist properties of Cys1-NC are associated with the presence of the sulfhydryl group of cysteine. The analogues D-Cys2 and Cys3 were also almost inactive.

[Current views on the etiopathogenesis of endometriosis]. / Wspólczesne poglady na etiopatogeneze endometriozy.

In the article contemporary knowledge concerning the ethiopathogenesis of endometriosis has been reviewed. The authors especially concentrate on immunological disorders in this disease, emphasizing the complexity of the development of endometriosis and multiple factors influencing its ethiopathogenesis.

Regulatory effects of lactoferrin and lipopolysaccharide on LFA-1 expression on human peripheral blood mononuclear cells.

The aim of this study was to investigate effects of human lactoferrin (hLF) with regard to LFA-1 expression on unstimulated and lipopolysaccharide (LPS)-activated human peripheral blood mononuclear cells (PBMC). The investigations were carried out on 30 healthy volunteers, males and females, 24-58 years old. We found that hLF, at an optimal dose of 5 microg/ml/10(6) cells in 24-hour culture, exerted regulatory effects on LFA-1 expression, depending on distribution of this molecule on cells in control cultures and on the effects of LPS. First, we revealed several patterns of LFA-1 distribution and density of this marker among studied individuals. The effects of LPS and hLF on LFA-1 expression patterns were differential. LFA-1 expression was stimulated by individual actions of LPS or hLF, additive or synergistic effects of both factors, it could be also inhibited by hLF alone or in combination with LPS. In about one third of cases no significant effects of LPS or hLF on LFA-1 expression were seen. Removal of monocytes from the PBMC population diminished LFA-1 expression in control cultures and abolished LPS- or hLF-elicited changes. The regulatory effects of hLF were also blocked by treatment of PBMC cultures with anti-tumor necrosis factor alpha (TNF-alpha) antibodies. Taken together, the data showed that hLF and LPS had immunoregulatory properties with respect to LFA-1 expression on human PBMC and that these actions were mediated by monocytes and TNF-alpha.

Oral treatment of rats with bovine lactoferrin inhibits carrageenan-induced inflammation; correlation with decreased cytokine production.

The aim of this study was to investigate an effect of oral treatment of rats with bovine lactoferrin (BLF) on carrageenan-induced inflammation. Rats were given 5 oral doses of BLF (10 mg each) on alternate days and 24 h after the last dose a carrageenan inflammation was induced in the hind foot. Control rats were given 0.9% natrium chloride (NaCl) or bovine serum albumin (BSA). The magnitude of the reaction was measured after 2 h (optimal response) and expressed as an increase of the foot pad thickness in milimeters. The evaluation of BLF effects on carrageenan reaction was supplemented by determination of the ability of spleen cell cultures to produce interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-alpha) upon lipopolysaccharide (LPS) induction using bioassays. The results revealed an inhibition of the carrageenan-induced inflammation in BLF-treated rats by 50 and 40% as compared to NaCl and BSA control groups, respectively. The inhibition was also associated with a substantial decrease in the ability of splenocytes to produce IL-6 in BLF-treated rats (94 and 83% as compared to NaCl- and BSA-treated groups). The LPS-induced TNF-alpha production was also decreased, although to a lesser degree (48 and 35%, respectively). The decreased ability of spleen cells to produce inflammatory cytokines in BLF-treated rats indicates that hyporeactivity of the immune system cells may be the basis for the inhibition of carrageenan-induced inflammation.

[Cytotoxic activity of NK cells in the presence of the peritoneal fluid fro women with unexplained infertility and with endometriosis related infertility]. / Aktywnosc cytotoksyczna komórek NK w obecnosci plynu otrzewnowego pobranego od kobiet z nieplodnoscia niejasnego pochodzenia oraz z nieplodnoscia zwiazana z endometrioza.

We examined peritoneal fluid obtained during laparoscopy in the aspect of its influence on cytotoxic activity of NK cells. We showed significant suppression of this activity in women with unexplained infertility. The suppression of NK activity was also present in women fertile and infertile both, with severe endometriosis. These results may indicate the immunological disorders in unexplained infertility and in endometriosis-related infertility.