The immunomodulating properties of a diphtheria bacterial vaccine in in vitro experiments

In this investigation lymphocytes sensitized with Corynebacterium diphtheriae antigens obtained from carriers and convalescents were used. The new diphtheria bacterial vaccine Codivac, in contrast to other comparable preparations (diphtheria toxoid, staphylococcal toxoid, staphylococcal vaccine, levamisole), was found to produce a more direct effect by modulating the levels of T-lymphocytes, depending on their initial levels in the patient. Codivac, together with other preparations, can be used for the study of the problems of immunostimulation and immunocorrective therapy.     

Multicomponent vaccine VP-4 in the therapy of allergic diseases

In the present study the results of the polycomponent vaccine B[symbol: see text]-4 use for the therapy of patients with bronchial asthma (BA) and latex allergy were generalized. The vaccine was introduced by the nasal-subcutaneous or nasal-oral administration simultaneously with the basic therapy. Te studies were conducted first on limited groups of patients, then in the course of the State Trial with the use of placebo control. Excellent and good effect lasting for 1 year and over was registered in 36 patients (66.7%) out of 54 BA patients receiving the vaccine by the intranasal-subcutaneous method. Immunotherapy produced no positive effect in 13 patients (24.1%). Out of 35 examined patients receiving the vaccine by the intranasal oral method, excellent and good effect was registered in 26 patients (74.2%). No effect was registered in 4 patients (11.4%). In the group of 28 patients receiving placebo simultaneously with the basic therapy positive dynamics in the course of the disease was observed only in 3 patients. Treatment with polycomponent vaccine B[symbol: see text]-4 led to a prolonged (to a year and more) decrease in the frequency and severity of exacerbations, contributed to the prolongation of remissions and to a decrease in the amount of administered medicinal preparations, especially systemic corticosteroids. Immunotherapy ensured the correction of the content of lymphocyte subpopulations with markers CD3, CD4, CD72 and a rise in the titers of antibodies to antigens contained in the preparation. The use of therapeutic polycomponent vaccine B[symbol: see text]-4 for the treatment of patients with latex allergy ensured the state of prolonged remission in this group of patients. On the basis of our investigations we believe that the use of the therapeutic polycomponent vaccine B[symbol: see text]-4 may be included into the basis therapy of allergic diseases.     

The reactogenicity and efficacy of a dried cell-free staphylococcal vaccine in treating chronic bronchitis]

The data obtained in the controlled trial of the immunotherapy of chronic bronchitis (CB) patients with lyophilized acellular staphylococcal vaccine developed at the Mechnikov Research Institute for Vaccines and Sera (USSR Acad. Med. Sci.), Moscow, are presented. The patients were divided at random into two groups; of these, one group received the vaccine and the other, placebo (0.9% sodium chloride solution). The preparations were injected subcutaneously simultaneously with traditional treatment. The vaccine was well tolerated by the patients. In the blood of the patients receiving the vaccine, in contrast to those receiving placebo, a significant increase in the level of specific antibodies determined in the passive hemagglutination test and IgG was noted. When following the remote consequences of the disease for a year after the course of immunotherapy, a significant decrease in the occurrence of aggravations, an increase in the duration of remission and a decrease in the duration of sick leaves were registered. These data indicate that the immunotherapy of CB with lyophilized acellular staphylococcal vaccine is a promising method of preventing relapses.     

Protective efficacy of combined administration of the multicomponent vaccine and the immunomodulator myelopid in experimental infections in mice

The influence of myelopid (MP) on the protective activity of polycomponent vaccine VP-4 prepared from the antigens of opportunistic bacteria was studied on experimental infections of mice, caused by Klebsiella pneumoniae, Salmonella typhimurium and Staphylococcus aureus. In staphylococcal and Klebsiella infections the joint administration of vaccine VP-4 and MP produced more pronounced protective effect than each of these preparations, introduced alone. The protective action of vaccine VP-4 was specially enforced by MP in cases of local staphylococcal infection. Recommendations on the joint use of two or more immunomodulating agents are possible only on the basis of the experimental substantiation of their effect in definite infections.     

Identification of vaccine candidate antigens of Staphylococcus aureus by serological proteome analysis

In this study we applied serological proteome analysis (Klade, C. S. et al. Proteomics 2001, 1, 890-898) for identification of bacterial vaccine candidate antigens. First, approximately one hundred sera from healthy individuals and patients suffering from Staphylococcus aureus infections were screened for antibodies against staphylococcal lysates and recombinant proteins representing surface antigens. Two pools (healthy donors, patients) each consisting of five sera with the highest antiproteinaceous IgG reactivity were selected. Second, S. aureus COL was grown under different conditions and the number of antigens expressed was monitored by Western blot analysis. Third, surface proteins were enriched by digesting the bacterial cell wall under isotonic conditions and subsequent removal of protoplasts. These protein preparations were resolved by two-dimensional electrophoresis (2-DE) (pI 4-7). 2-DE immunoblotting using the preselected serum pools at 1:10 000-1:100 000 dilutions revealed a number of highly immunogenic staphylococcal proteins. Twenty-one spots were isolated by preparative 2-DE, and analysed by matrix-assisted laser desorption/ionization mass spectrometry and tandem mass spectrometry sequencing of tryptic peptides. This led to the identification of 15 proteins including known and novel vaccine candidates. Seroreactivity of several antigens including serine-aspartate repeat containing protein D, immuno-dominant staphylococcal antigen and a novel 309 amino acid lipoprotein was independently confirmed by enzyme-linked immunosorbent assay and Western blot analysis of purified recombinant proteins. In conclusion, serological proteome analysis proved to be a powerful tool for the identification of novel staphylococcal antigens, which provide a basis for rational vaccine design.     

The effect of repeated administration of staphylococcal immune preparations on the development of local staphylococcal infection in mice

The study is concerned with the effect of repeated administration of staphylococcal immunopreparations on the development of a suppurative-inflammatory focus in the foot of the mouse. Subcutaneous administration of large doses of the antigenic complex of the staphylococcus (ACS) obtained by aqueous extraction, antiphagin and native anatoxin failed to induce an increase in sensitivity to staphylococcus. In some cases, the extent of development of the suppurative-inflammatory focus in the mice which had been given these preparations was less than in the control; this is suggestive of their protective effect. When comparing, on this model, the ACS preparations and corpuscular vaccine produced from poorly and highly virulent strains, we observed a more pronounced protective effect in the preparations from the poorly virulents strains. The extent of oedema was greater than in the control when adsorbed anatoxin was administered. The administration of staphylococcal preparations with a therapeutical purpose after staphylococcus infection caused a significant decrease in the size and intensity of manifestation of the suppurative-inflammatory focus in the foot. The model of limb oedema enabled us to reveal the sensitizing and protective effect of the preparations under study.     

Assessment of the immune status of chronic bronchitis patients in the remission phase during prophylactic treatment with levamisole and staphylococcal vaccine

The work presents the results of the study of the immune status in 18 patients with chronic bronchitis at the stage of remission, subjected to prolonged prophylactic treatment with the inhalations of levamisole solution, and in 16 patients receiving the inhalations of staphylococcal vaccine. These results indicate that the inhalations of levamisole have proved to be more effective than the inhalations of staphylococcal vaccine. In patients treated with levamisole an essential decrease in the ratio of theophylline-resistant to theophylline-sensitive E-rosette-forming lymphocytes has been noted. The determination of the sensitivity of lymphocytes and neutrophils to incubation with levamisole by means of the rosette-formation test makes it possible to prognosticate, taking into account the initial level of spontaneous E-rosette formation of these cells, the effectiveness of the prophylactic treatment of patients with chronic bronchitis. The determination of the sensitivity of the cells to incubation with staphylococcal toxoid is ineffective for the prognostication of the result of the treatment of such patients with staphylococcal vaccine.     

Prevention and immunotherapy of staphylococcal infections with bacterial vaccines

Pathogenesis of staphylococcal infection both local and systemic is associated with many pathogenicity factors, which in foreign literature are called virulence factors of Staphylococcus aureus, which were studied as potential candidates for vaccine development. Much difficulties are related to use of known experimental models, which virtually do not allow to determine direct appropriate effect by survival of animals, as well as to data about absence of correlation between increase of antibody titers in animals and protective effect of studied preparations. Despite the importance of the problem of prevalence and severity of staphylococcal infection and intensive research in order to determine protective components able to protect from infection caused by S. aureus, there are no licensed prophylactic preparations with proven efficacy.     

Optimization of the therapy of atopic dermatitis by means of immunotherapy

Taking into account disturbances in the functioning of the immune system in atopic dermatitis (AD) and the potentiating role of staphylococcal and other infections, the possibility of the optimization of the therapy of AD with the use of preparations having immunomodulating action and immunogenic activity is proposed. In the complex therapy of AD in children we used polycomponent vaccine Immunovac B-4, introduced intranasally and orally. Under the influence of immunotherapy the clinical characteristics of the patients had pronounced positive dynamics. A considerable decrease in the spread of the process, the degree of its severity and subjective symptoms was noted shortly after the course of vaccine treatment. Simultaneously the SCORAD index dropped from 64.5 to 39.4. During the later period of observation further decrease in the severity of the course of AD in children occurred, and the minimal characteristics were observed in 6 months of observation. At that time the SCORAD index fell to 19.9 +/- 1.34. The volume of pharmacotherapy and the number of acute respiratory infections considerably decreased, the positive dynamics of the quantitative and qualitative composition of the intestinal microflora was noted. The prolonged immunotherapeutic effect of the polycomponent vaccine made it possible to recommend the vaccine for the optimization of the therapy of AD.     

Experimental investigation of preparations for the immunotherapy of staphylococcal infections

A comparative experimental study was carried out of antigenic staphylococcal preparations developed in the USSR and Czechoslovakia for the immune therapy of chronic staphylococcal infection. The efficiency of the preparations was unequivocally confirmed using the rabbit staphylococcal sepsis model. The immunogenicity of tested strains was shown not to always correlate with their virulence. Preparations obtained by means of aqueous extraction from mildly virulent immunogenic strains exhibited greater protective activity than those prepared from highly virulent strains. The PCA phenomenon did not differ significantly provided the tested preparations were administered at doses which ensured equal protective action.     

Use of a lysoenzyme preparation from Streptomyces recifensis subsp. lyticus 2435 for isolating DNA from staphylococcal cells

It was shown that the preparation 2435 from Streptomyces recifensis subsp. lyticus, including a complex of bacteriolytic and concomitant enzymes provided lysis of thick staphylococcal suspensions within 15 to 20 minutes under optimal conditions after preliminary treatment of the cells with 0.1 M cystein-HCl. A procedure was developed for isolating DNA from the cells of staphylococci and other microorganisms based on enzymatic lysis. In terms of major physicochemical properties, the preparations of DNA were not inferior to the preparations of DNA isolated by the classical Marmur technique with Kirbi's deproteinization and had transforming activity. The developed procedure for isolation of DNA with using the lysoenzyme preparation widened the possibilities of investigating the genetics of staphylococci and other microorganisms.     

Effect of antibiotic therapy and vaccine therapy on the phagocytic reaction in mice infected with Staphylococcus

The time course of changes in the activity, intensity and completeness of phagocytosis with leukocytes of the peritoneal exudate was studied on mice with experimental staphylococcal infection treated with rifampicin, lincomycin and inactivated staphylococcal vaccine used alone or in combination. It was shown that immunization of the animals with inactivated staphylococcal vaccine promoted stimulation of the phagocytic defense. Rifampicin and lincomycin applied therapeutically induced a decrease in the activity, intensity and completeness of phagocytosis. It should be noted that rifampicin had a less pronounced inhibitory effect than lincomycin. The combined use of vaccine and antibiotics with therapeutic purposes promoted an increase in phagocytosis as compared to the use of the antibiotics alone. The combined therapy sometimes resulted in completeness of phagocytosis making it reach the control values (the 10th and 15th days, rifampicin and vaccine). It should be noted that a more pronounced stimulation of the activity, intensity and completeness of the phagocytosis was observed with the use of the combination of rifampicin and the vaccine.     

The comparative characteristics of the immunomodulating properties of protein A from Staphylococcus aureus of different origins

The immunomodulating properties of highly purified staphylococcal protein A and its analog obtained by gene engineering techniques have been compared with those of commercial preparations. The comparison has shown that the differences observed in this investigation may be explained by the presence of admixtures of staphylococcal nature in commercial preparations. The preparations of highly purified staphylococcal and recombinant protein A stimulate humoral immune response and the processes of phagocytosis and do not show mitogenic activity with respect to T cells. The conclusion on the identity of the immunomodulating activity of the preparations of natural and recombinant protein A has been made.     

Immunological activity of a Proteus vaccine administered to volunteers simultaneously with a pyoimmunogen and adsorbed staphylococcal anatoxin

The comparative study of the immunological activity of Proteus vaccine prepared from soluble antigenic complexes was made after the immunization of volunteers with this vaccine used in the form of a single preparation and in combination with pyoimmunogen (Pseudomonas aeruginosa vaccine) and/or adsorbed staphylococcal toxoid. The injection of the vaccine in the form of a single preparation and in different combinations increased the ingestion of Proteus cells by neutrophils. The injection of Proteus vaccine simultaneously with pyoimmunogen and staphylococcal toxoid ensured the intensive phagocytosis of staphylococci. All combinations with Proteus vaccine, used in this investigation, stimulated the intensive formation of antibodies to Proteus vaccine strain and Re-glycolipid. Proteus vaccine introduced in combination with adsorbed staphylococcal toxoid essentially stimulated the synthesis of anti-alpha-staphylolysin.     

The methods for correcting with specific and nonspecific immunomodulators the immunopathological processes arising from the administration of a staphylococcal vaccine]

In mice immunized with staphylococcal vaccine the arresting of graft-versus-host reaction under the influence of small doses of staphylococcal vaccine, hyperimmune antistaphylococcal serum, cyclophosphamide, antilymphocytic serum has been demonstrated. Small doses of staphylococcal vaccine stimulated the production of antibodies to staphylococci and dermal extract in the animals, previously immunized with this vaccine, with the simultaneous suppression of cell-mediated immune reactions to both antigens. Immunosuppressing agents have been found to inhibit humoral and cell-mediated immune response to microbial antigen and dermal extract. No influence of vermox and levamisole on the outcome of the graft-versus-host reaction has been registered; the latter preparation has been found to intensify cell-mediated immune reactions to microbial and tissue antigens.     

Effect of lincomycin and staphylococcal vaccine on the course of experimental staphylococcal sepsis

Therapeutic efficacy of lincomycin used alone and in combination with inactivated staphylococcal vaccine and the effect of these agents on synthesis of antibodies and their content in blood serum were investigated. Lincomycin was shown to inhibit septic processes in the host. After its administration the number of the pathogens in the blood and organs markedly decreased. At the same time, lincomycin lowered antibody synthesis in the lymphoid organs and the content of alpha-antitoxins in blood serum. The use of lincomycin in combination with inactivated staphylococcal vaccine promoted an increase in the number of the antibody forming cells in the spleen and lymph nodes and the content of the antibodies to the staphylococcal alpha-toxin in blood serum of the animals with staphylococcal sepsis.     

Effect of antibiotic and vaccine therapies on the succinate dehydrogenase and phosphatase activity of liver cells of mice infected with Staphylococcus

Changes in the activity of succinate dehydrogenase (SDH), total and acid phosphatase (TP and AP) were studied in treatment of laboratory animals with rifampicin, lincomycin and with inactivated staphylococcal vaccine used alone or in combinations. It was shown that immunization of the animals with inactivated staphylococcal vaccine under conditions of experimental staphylococcal infection promoted stimulation of the enzyme activity. Rifampicin and lincomycin used for the treatment of such animals lowered the activity of the enzymes. The suppressing effect of the antibiotics increased with an increase in the period of their use. It should be noted that the inhibitory effect of rifampicin on the activity of SDH, TP and AP was less pronounced than that of lincomycin. The combined use of the vaccine and antibiotics for the treatment of the animals promoted an increase in the enzyme activity as compared to the use of the antibiotics alone. Sometimes the activity of SDH, TP and AP reached the control levels in such animals or the levels observed in the animals treated with the vaccine alone. Stimulation of the enzyme activity was more pronounced when the vaccine was used in combination with rifampicin.     

Use of therapeutic polycomponent vaccine "Immunovac VP-4" for the treatment of chronic bacterial prostatitis

A complex of clinical and laboratory examinations of patients with bacterial chronic prostatitis was carried out. The patients were divided into two groups: 55 patients treated by the method of immunotherapy and 20 patients (controls) undergoing basic therapy. Immunotherapy was carried out with the use of polycomponent vaccine VP-4 containing Staphylococcus aureus, Klebsiella pneumoniae, Proteus vulgaris and Escherichia coli antigens. Vaccinal therapy was indicated in cases with infection focus associated with opportunistic bacteria, a prolonged torpid course of prostatitis and when different schemes of etiotropic therapy proved to be ineffective. The vaccine was introduced in 4 or 5 subcutaneous injections at an interval of 5-7 days. The study revealed that in cases of chronic prostatitis accompanied by microecological and immune disturbances the use of complex therapy allowed to achieve good clinical and laboratory results in 91% of the patients. In the patients undergoing immunotherapy the elimination of opportunistic bacteria from the urogenital tract and the restoration of affected microflora occurred.     

Immunological reactivity of patients with acute infectious destruction of the lungs in vaccination with a staphylococcal vaccine

The work presents the results of the study of the immune status in 32 patients with acute infectious destructions of the lungs; of these, 17 patients were immunized with staphylococcal vaccine, which made it possible to achieve a positive clinical effect in 16 patients, thus shortening their sick-leave. The injection of the preparation was shown to increase the number of E-rosette-forming cells, "active" E-rosette-forming cells, theophylline-resistant and theophylline-sensitive lymphocytes.     

Effect of an autovaccine on the course of an experimental staphylococcal infection]

The effect of autovaccine on the state of cellular immunity in mice with staphylococcal infection was studied. The maximum decrease of staphylococcal dissemination in internal organs, espeically in the lungs, as well as an increase in the intensity of phagocytosis by peritoneal macrophages were observed after the administration of the vaccine by the method of inhalation. The intranasal administration of the vaccine also proved to be more effective than subcutaneous injection. The cumulation of immune response was more pronounced after the aerosol administration of autovaccine, especially in cases of pathological processes in the respiratory organs.