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[Comparative studies of the paraspecific immunostimulation (paramunization) by bacterial lysates in bacterial infection models and in the cytotoxicity test].

Bacterial lysates of different bacterial strains (E. coli, B. bronchiseptica, P. haemolytica) were prepared by heating, acid- and alkaline-hydrolysis. Lysates were tested for their immunostimulating effect in bacterial infection models and with chromium 51 test demonstrating spontaneous (natural) cytotoxicity. Lysate production was standardized by protein- and Lps-determination. The alkaline-hydrolysis reduced toxicity of Lps and increased the content of soluble bacterial protein. Heating and acid-hydrolysis did not alter bacterial suspensions with respect to Lps-toxicity and protein-content. Mice infected with P. aeruginosa, P. multocida, E. coli and L. monocytogenes (5-10 LD50) had a significantly longer survival time after prophylactic immunostimulation with bacterial lysates than control animals. No protection was observed in immunostimulated mice infected with Erysipelothrix rhusiopathiae. In the Pseudomonas infection model, bacterial lysates prepared by alkaline-hydrolysis had a 10 times higher immunostimulating effect than lysates prepared by acid-hydrolysis or heating. Bacterial lysates stimulated spontaneous cytotoxicity of natural mouse peritoneal killer cells after intraperitoneal application. Whole bacterial lysates had a higher NK-activity as their corresponding purified lipopolysaccharide portion.

Adjuvants, Immunologic↗

Multiplex PCR for screening of integrons in bacterial lysates.

Bacterial integrons are a useful PCR amplification target in epidemiological surveys of bacterial antibiotic resistance, and a variety of primers have been published. We describe multiplex PCR methodology to test for classes 1, 2 and 3 integron-associated integrases in boiled lysates of Gram-negative bacteria. We report on performance in Acinetobacter spp. (n=50), Enterobacteriaceae (n=76), Pseudomonas aeruginosa (n=15), Bacteroidesspp. (n=69), and in undifferentiated mixed cultures derived from perineal swabs (n=50) and endotracheal aspirates (n=8). This method achieved 100% sensitivity and specificity in simple lysates made from a range of bacteria, without requiring DNA extraction, and is recommended as an efficient screening tool for surveys of integron cassettes.

Acinetobacter↗

[Interferon induction in lymphocytes using a bacterial lysate].

A polyvalent bacterial lysate (IRS 19) could be shown to induce interferon (IFN) in peripheral mononuclear leukocytes isolated from human and mouse blood by density gradient centrifugation. Upon stimulation with optimal doses of the inducer, the cells of both species produced 1000 to 2000 IU IFN/ml when cultured for up to 72 hours. In both systems the interferon was released to the culture supernatants in a first wave from one to four hours after addition of the inducer and in a second beyond the 16th hour of incubation increasing up to 72 hours, this time, reaching a 10 fold higher titer.

Animals↗

Open comparative, randomized controlled clinical study of a new immunostimulating bacterial lysate in the prophylaxis of upper respiratory tract infections.

One hundred and fourteen patients with a medical history of recurrent upper respiratory tract infections (at least four episodes in the year preceding this open comparative study) were randomly assigned to three groups of 38 patients each. The first group received, by sublingual route, Ismigen, a new immunostimulating lysate (Polivalent Mechanical Bacterial Lysate, PMBL) obtained by mechanical lysis of 48 billion bacteria commonly responsible for upper respiratory tract infections; the second group received an oral immunostimulating lysate (CLBL) obtained by chemical lysis of 36 billion bacteria. A third group served as control and did not receive any immunostimulating treatment (Control NT). One tablet a day of PMBL was given to the first group for the first ten days during three consecutive months; the patients of the second group received one capsule a day of CLBL with the same treatment schedule. At the end of the treatment period the patients of the 3 groups were followed up for 3 months. The primary end point was the number of acute upper respiratory tract infections (URTIs) that occurred during the three months of treatment and three months of follow-up. Secondary endpoints were: the number of patients free from disease, the duration of infectious episodes, the number of working days lost because of disease, the need for a concomitant antibiotic treatment, and the safety of the two treatments. During the treatment period the mean number (+/- SD) of URTIs per patient was 0.34 (0.48) in the PMBL group, 1.0 (0.83) and 1.23 (0.77) in the CLBL and Control NT groups, respectively. Results of PMBL treatment were significantly better (p < 0.05) than the results in the other two groups; CLBL was not significantly different from the control group. In the three months of follow-up, the mean number (+/- SD) of URTIs per patient was: 0.42 (0.55) in the PMBL group, 0.92 (0.67) in the CLBL group, and 1.55 (0.88) in the Control NT group. The PMBL group was significantly better than the other two (p < 0.05). During the 6-month study, significantly more patients of the PMBL group remained free from respiratory infections in comparison to the other two groups. As regards other secondary end points (duration of infectious episodes and number of working days lost), the mean values of the PMBL group were statistically significantly lower than those of the other two groups, in both the treatment and follow-up periods. No PMBL treated patient needed concomitant administration of an antibiotic, while 9 patients of the CLBL group received such treatment (p < 0.05). No patient reported adverse events.

Adjuvants, Immunologic↗

Effect of oral bacterial lysates on serum immunoglobulins.

The level of serum immunoglobulins IgG, IgA, IgM and IgE has been studied before and after oral immunotherapy with a bacterial lysate in 88 patients with bronchial asthma, repeated respiratory infection and 12 cases of IgA deficiency. A significant increase in IgA has been observed in 9 patients presenting initially a decreased IgA serum level. In 3 patients without response to the standard treatment an increase in IgA was achieved increasing the dosage of oral bacterial lysate. Oral bacterial lysates could be an useful immunomodulating agent in repeated respiratory infections associated or not with IgA deficiency.

Adjuvants, Immunologic↗

[Multicenter study of the immunostimulating activity of a bacterial lysate].

The Authors report the results obtained within a multicentrical trial, weighing the immunostimolant effect of bacterial lysate on 157 patients. The drug (bacterial lysate) has induced an immunitary reaction, making significantly higher either the salivary IgA values or the serum IgA, IgG and IgM values. Also excellent tolerability is peculiar to this new immunostimolant.

Adjuvants, Immunologic↗

Orally administered bacterial lysate Broncho-Vaxom for the treatment of common variable immunodeficiency.

Broncho-Vaxom (B-V) is a lysate of eight bacterial pathogens of the respiratory tract with immunomodulatory properties. It is used in prophylaxis of respiratory tract infections. We conducted an open, placebo controlled, cross-over trial of B-V treatment in patients with common variable immunodeficiency (CVID). B-V or placebo were given for the period of four months either in winter or spring period. No significant improvement of clinical state of patients during the period of B-V treatment compared with the period when placebo was administered was observed. When subjective comparison of the health state of patients with the same period of the previous year was assessed, significant improvement after B-V treatment compared to placebo was recorded. Evaluating serum immunoglobulin levels a significant increase in serum IgA level was observed after B-V treatment. The results warrant further studies of B-V treatment in CVID patients.

Adjuvants, Immunologic↗

Enhanced lymphocyte stimulation by bacterial lysates after treatment of probands.

The proliferative response of lymphocyte cultures upon the addition of a mixture of antigens from the lysates of different bacteria (Paspat) to the culture medium was investigated using lymphocytes of groups of probands that investigated using lymphocytes of groups of probands that had been treated with the i.c. test preparation (T.P.1, group I), the lyophilized test preparation given orally (T.P.2, group II), a commercially available lyophilized bacterial lysate of similar composition given orally (T.P.3, group III), or with placebo (group IV). Lymphocyte cultures were set up on day 0 (before treatment), day 40 and day 70 (after treatment). The results show, that T.P.1 and T.P.2 produce a dose dependent proliferative response in lymphocyte cultures with a peak reactivity between 70 and 210 micrograms/ml. The degree of stimulation obtained with the bacterial lysate in different concentrations is increased in groups I and II over the stimulation obtained before treatment. Groups I and II which were treated with T.P.1 (i.c.) and T.P.2 (orally) are significantly different in their response to the bacterial lysate from the groups treated with T.P.3 or placebo on days 40 and 70. Evidence is presented that the stimulation obtained is predominantly a proliferation of T-cells.

Antigens, Bacterial↗

Bacterial lysates and ribosomes as inducers of specific immune responses: a comparative study.

A bacterial lysate (OM-85 BV), a preparation of purified bacterial ribosomes (D53) and a placebo were tested for ability to induce the local appearance of specific antibody-containing cells. The three compounds were given orally to 90 children who required tonsillectomy. Surgery was carried out after 1 month of therapy. Frozen-cut sections of each tonsil were tested in indirect immunofluorescence. Cells containing antibodies directed to Streptococcus pneumoniae, Streptococcus pyogenes, Haemophilus influenzae or Klebsiella pneumoniae were enumerated. Lowest values were noted in the placebo group. Slightly higher numbers were observed after treatment with OM-85 BV, but significant increases were noted only for the elevated numbers of specific antibody-containing cells observed after D53 therapy. Bacterial ribosomal preparations thus contribute efficient induction of specific local immune responses in man.

Adjuvants, Immunologic↗

[Prospective placebo-controlled double-blind study using a bacterial lysate in infections of the respiratory tract and ENT region in children].

94 children suffering from frequent infections of the respiratory tract and of the ear, nose and throat were treated under double-blind conditions with either a bacterial lysate (n = 45) or a placebo (n = 49). During the 6 months of the trial both treatments brought about a significant decrease in the incidence and duration of these infections as well as in the duration of concomitant antibiotherapy in comparison to the corresponding prior 6-month reference period. As these positive results recorded under the bacterial lysate and the placebo could not be differentiated statistically, the influence of meteorological and epidemiological factors as well as of the age of the children is discussed.

Adjuvants, Immunologic↗

Oral immunization with polyvalent bacterial lysate and infection with Streptococcus pneumoniae: influence on interferon-gamma and PMN elastase concentrations in murine bronchoalveolar lavage fluid.

We recently demonstrated that oral immunization with a polyvalent bacterial lysate (Paspat oral) significantly reduces mortality rates in mice, infected with Streptococcus pneumoniae or influenza A virus. In this study it is demonstrated that oral immunization with the same bacterial lysate reduces the intrapulmonary inflammatory reaction to infection with S. pneumoniae, assessed by measurement of PMN elastase in bronchoalveolar lavage fluid. Furthermore, it is demonstrated that oral immunization with Paspat oral increases intrapulmonary IFN-gamma concentrations.

Administration, Oral↗

An open-label, prospective study of an oral polyvalent bacterial lysate (Luivac) in the treatment of recurrent respiratory tract infections in Thai patients.

An open-label, non-comparative study was performed in the Department of Otolaryngology, Siriraj Hospital, Bangkok, Thailand, to assess the safety, tolerability, acceptability and efficacy of an oral polyvalent bacterial lysate (Luivac) in the treatment of recurrent respiratory tract infections (RTIs) in Thai patients. Thirty-three patients were included in this study, 18 males and 15 females, with a mean age of 34.0 +/- 14.7 years. The mean number of RTIs during the 12-month period preceding the study was 9.5 per patient. During the study each patient received one tablet of Luivac daily for 28 days followed by a treatment-free period of 28 days. This was followed with another 28 days on Luivac, after which there was a 28-day treatment-free follow-up period. This study lasted 4 months with five scheduled patient visits (V1-V5). Laboratory studies were done at baseline (V1) and after treatment (V4), which included complete blood count and serum immunoglobulins (IgA, IgE, IgG and IgM). The incidence of all adverse events was 15.2% and no case was related to the studied drug. There were no clinical relevant changes in laboratory parameters after treatment. The reduction rate of RTIs per month at the end of the study period was 63.5% when compared to the average RTIs rate per month during the 12 months preceding the study. A comparison of the first study period (V1-V3) and the second study period (V3-V5) showed a reduction in duration of RTIs (23.1%), in the clinical infection score (17.5%), in the number of antibiotics used (2.1%), in the number of symptomatic treatments (3.5%), and in the number of days absent from school or work (50.0%). Overall tolerability and acceptability were assessed as very good and good in 96.8% of the patients. This study suggests that oral polyvalent bacterial lysate (Luivac) was safe and also showed a tendency to be effective in preventing RTIs in Thai patients with or without risk factors for recurrent RTIs. Other clinical advantages were reduction in the severity and duration of infection as well as in reduction of the cost of treatment and the number of days absent from school or work.

Administration, Oral↗

Immunomodulation of allergic autocytotoxicity in bronchial asthma by a bacterial lysate--Broncho-Vaxom.

The direct and antibody-dependent allergic autocytotoxicity (ACT) response, mediated by food antigens and its immunoregulation with bacterial lysate of the eight most common pathogens of the upper respiratory tract--Broncho-Vaxom (BX), was investigated in fifteen bronchial asthma patients and eight normal control individuals. Under the described experimental conditions, the BX inhibits ACT response in vitro. In analyzing the mechanism of this effect, the enhancement of T suppressor cells by BX was under consideration.

Adjuvants, Immunologic↗

In vivo effect of an immunostimulating bacterial lysate on human B lymphocytes.

The aim of the present study is to investigate in humans the mechanism by which the oral vaccine Polyvalent Mechanical Bacterial Lysate (PMBL) can rapidly mobilize specific immune response and evaluate the efficacy of its immunostimulating activity in preventing recurrent infections of the upper respiratory tract (URTIs) in a group of patients with a medical history of URTI recurrence. Patients received, by sublingual route, PBML, an immunostimulating lysate obtained by mechanical lysis of the most common bacteria responsible for upper respiratory tract infections. The treatment was administered for 10 consecutive days/month for 3 consecutive months. After the end of the treatment period the patients were followed up for an additional 3 months. The frequency of IgM memory B cells and the expression of the activation marker CD25 in peripheral blood lymphocytes were measured using the flow cytometric method before the start and at days 30 and 90 of the treatment cycle. To correlate clinical results to immunological parameters, the patients were monitored at different time-points during the treatment and at the end of follow-up period. The results showed that PMBL exerts a therapeutic and preventing effect in acute and recurrent infections of the upper respiratory tract and that this effect correlated with the activation and enhancement of both IgM memory B lymphocytes (CD24+/CD27+ cells) and IL2 receptor-expressing lymphocytes (CD25+ cells) involved either in humoral or cellular immunity.

Adjuvants, Immunologic↗

Protection against influenza A virus infection in mice by oral immunization with a polyvalent bacterial lysate.

A study is presented which investigated whether oral immunization with a polyvalent bacterial lysate (Paspat oral) can sufficiently enhance cell-mediated defense mechanisms to protect mice against influenza A virus infection. It was found that oral immunization reduced mortality due to influenza A infection with 15-70%, depending on the quantity of virus administered and and the moment of infection. Cyclosporin A severely reduced the protective effect of oral immunization, suggesting that a major effect of oral immunization in these studies is T-cell activation. The effect of oral immunization on macrophageal activity was evaluated by measuring cyclic-AMP in alveolar macrophages (AMs) obtained by bronchoalveolar lavage. Before infection, basal activity levels of AMs in immunized mice were significantly lower than in controls. Five days after infection, however, basal activity level of AMs in immunized mice was significantly higher than AM activity in controls. Stimulation of AMs with PGE2 significantly reduced cellular activity in both groups, before and after infection. However, cellular activity of AMs from immunized animals was less reduced than cellular activity of control macrophages. Activity of AMs of immunized animals was significantly more reduced by histamine than activity of control macrophages. It is concluded that oral immunization with Paspat oral stimulates T-cell-dependent immune mechanisms, resulting in protection against influenza A virus infection in mice.

Administration, Oral↗

Protective effects of orally administered, Klebsiella-containing bacterial lysates in mice.

The efficacy, as oral vaccines, of hepta- and mono-valent, Klebsiella-containing bacterial lysates and a number of control preparations was tested in mice. The preparations were administered during two periods of four days each, interrupted by an interval of 3 days. Fourteen days after the first dose, the animals were challenged either intraperitoneally (i.p.; peritonitis/sepsis model) or intranasally (i.n.; pneumonia model). Animals treated with low doses of Klebsiella lysate, in the form of either a 7-valent lysate or a Klebsiella monolysate, showed enhanced survival in both the peritonitis/sepsis and the pneumonia models. Hexa- and tetra-valent preparations without Klebsiella were not protective in the models tested. Furthermore, it was found that the protection is accompanied by priming for Klebsiella-specific IgG responsiveness (probably at the T cell level) and by significant IgA anti-Klebsiella serum antibody levels in about one third of the animals. The oral efficacy of Klebsiella-containing lysates suggests the presence of an adjacent component that directs Klebsiella antigen(s) to follow a selective intestinal pathway which renders them immunogenic. The identity of this component is under investigation.

Administration, Oral↗