Photodynamic Inactivation of Mycobacterium tuberculosis Using Alluminium Phthalocyanine.

In a series of in vitro experiments, the optimum regimes of laser treatment were determined for effective photodynamic inactivation of Mycobacterium tuberculosis at a constant dose of aluminum phthalocyanine. Reference laboratory drug-susceptible strain H37Rv and clinical isolates of M. tuberculosis with varying degrees of resistance to antibiotics were used. Suspensions of M. tuberculosis were incubated with aluminum phthalocyanine in a concentration of 5 µg/ml and then subjected to photodynamic inactivation with high- or low- intensity laser irradiation at λ=662 nm at various parameters of light power density. Mycobacteria survival rate was assessed by CFU assay on solid media. It was shown that at the specified dose of the photosensitizer, the photodynamic inactivation of mycobacterium was characterized by inhibition and complete cessation of their growth depending on the dose density of the laser energy. Effective photodynamic inactivation started from a light dose density of 46.9 J/cm2 at a radiation power of 0.01 W and from 56.25 J/cm2 at a radiation power of 0.1 W. Photodynamic inactivation at low laser power is more effective against drug-susceptible strains of M. tuberculosis.

[Efficiency of lymphotropic intermittent chemotherapy in the complex treatment of patients with progressive pulmonary tuberculosis].

The study has established that during a progressive tuberculous process, immunological changes appear as significant increased proinflammatory cytokine production that leads to progressive destructive processes in the lung parenchymal lesion areas. Inclusion of lymphotrophic chemoimmunotherapy with roncoleukin into the complex therapy of patients with progressive pulmonary tuberculosis permits the optimization of the results of treatment in patients with progressive pulmonary tuberculosis due to the target delivery of antituberculous and immunoactive agents.

[Characteristics of A-interferon-generated dendritic cells in patients with pulmonary tuberculosis].

The phenotype of the dendritic cells (DC) generated from the adhesion fraction of mononuclear cells in the presence of GM-CSF and alpha-interferon was studied in patients with pulmonary tuberculosis. Despite the absence of significant differences in the count of mature CD83+DCs in the groups of patients (n = 38) and healthy donors (n = 30), elevated CD14(+)-monocyte levels and few activated CD25(+)-DCs were indicative of the impaired process of DC maturation/generation in patients with pulmonary tuberculosis, particularly in a subgroup of patients with a low T-cell proliferative response against PPD (PPD-anergy, n = 10). The patients with tuberculosis showed the lower relative levels of CD11c(-)-CD123(+)-DC and the normal levels of myeloid CD11c(+)D123(-)DCs. However, in patients with PPD-anergy, the content of myeloid CD11c(+)CD123(-)-DCs was significantly higher than that in PPD-reactive patients. Moreover, the patients with PPD-anergy were characterized by the elevated peripheral blood levels of CD14+CD16(+)-monocytes, which was associated with the high suppressive activity of monocytes (r(s) = 0.53; p < 0.05). The impaired process of DC generation/maturation in patients with pulmonary tuberculosis is believed to be associated with the changes in the phenotypic and functional properties of monocytes and to be a cause of an inadequate antigen-specific response in tuberculous infection.

Autologous bone marrow cells in the treatment of cirrhosis of the liver.

The data characterizing tolerance and efficiency of autologous bone marrow cells in the treatment of patients with cirrhosis of the liver are presented. Injection of autologous bone marrow cells was not associated with the development of adverse reactions. Cell therapy of patients with compensated cirrhosis arrested asthenic syndrome, reduced cytolysis, increased the level of serum albumin and platelet count. Ultrasonic examination revealed reduction of portal hypertension (the area of the spleen and the portal vein lumen decreased). In patients with decompensated cirrhosis, a positive response presenting as reduction of the disease severity (by 1.9 points) was observed in 48.6% cases. Positive shifts in these patients were associated with a decrease of ALT and AST levels, reduction of laboratory signs of cirrhosis, increase in platelet count, and reduction of the asthenic syndrome. Hence, therapy with autologous bone marrow cells is safe and, according to preliminary results, can be regarded as a new approach to the treatment of patients with cirrhosis of the liver.

Use of interferon-alpha-induced dendritic cells in the therapy of patients with malignant brain gliomas.

Clinical and immunological analysis of the efficiency of combined immunotherapy with the use dendritic cells for the treatment of malignant glioma of the brain was carried out. Dendritic cells generated in the presence of granulocyte-macrophage CSF and IFN-alpha retain their functional characteristics in patients with gliomas, which suggests the possibility of their use for the treatment of malignant tumors (glioma) of the brain. Combined therapy using interferon-induced dendritic cells was associated with generation of antigen-specific immune response during vaccinations. The results indicate satisfactory tolerance of combined immunotherapy using dendritic cells and the absence of toxic side effects at the stage of adoptive immunotherapy and at the stage of vaccinations with dendritic cells. Clinical trials showed that vaccinations with dendritic cells included into combined immunotherapy improved the quality of life and survival of patients with malignant gliomas.

[Interleukin-2 in the correction of T-cell anergy in patients with pulmonary tuberculosis].

The clinical and immunomodulating effects of lymphotropic administration of interleukin-2 (IL-2) were studied in the combine treatment of patients with pulmonary tuberculosis. The patients with tuberculosis were shown to have the low levels of monocytes with the intracellular expression of tumor necrosis factor-alpha (TNF-alpha) and the high count of CD14+ CD16+ monocytes with the intracellular expression of IL-10. The changes in the monocytic link were most pronounced in patients with PPD-induced anergy appeared as the low proliferation and production of alpha-interferon (alpha-INF). During clinical trials, 19 patients received tuberculostatic therapy in combination with IL-2 (Roncoleukin) (a study group) whereas 16 patients had tuberculostatic therapy alone (a control group). The administration of Roncoleukin statistically significant increased a proliferative response to PPD and normalized the count of CD14+ CD16+ monocytes with anti-inflammatory and immunosuppressive activities. The restoration of a PPD response was recorded more frequently in the study group than in the control one (75% vs 30%; p = 0.045). The magnitude of positive X-ray changes was also higher in the study group than in the control one (63% vs 25%; p = 0.026). The findings suggest the clinical and immunomodulating effects of IL-2 (Roncoleukin) in the combined therapy for tuberculosis.

[T-cellular energy deficiency in the pathogenesis of immunodeficiency in pulmonary tuberculosis]. / T-kletochnaia anergiia v patogeneze immunnoi nedostatochnosti pri tuberkuleze legkikh.

The phenotypic and functional properties of T cells were evaluated in patients with tuberculin anergy and a possible role of anergic T cells in the development of immune deficiency in pulmonary tuberculosis (PT) was studied. The profound decrease (more than 50%) depressed T-cell proliferation in PPD-stimulated cultures was shown to be recorded in 44% (59/134) patients with PT. PPD hyporesponsiveness was associated with the low proliferation of anti-CD3 and SEB-stimulated proliferation of a healthy donor's mononuclear cells evidencing the enlargement of anergic T cells with a suppressive activity in PT. A PPD-stimulated response in healthy donors was under the negative control of CG25-positive cells. In the PPD-anergic patients, there was a significant increase in CD4+CD25+ T cells that were inversely correlated with the PPD-induced proliferative response. The development of tuberculin anergy was more pronounced in patients with drug resistance. The intensity of a PPD-stimulated response in patients with tuberculin anergy may be restored in the presence of exogenous interleukin-2.

[Antigen-specific immunotherapy as a component of combined therapy for malignant brain tumors]. / Antigenspetsificheskaia immunoterapiia v kompleksnom lechenii bol'nykh zlokachestvennymi opukholiami golovnogo mozga.

The purpose of the present research was to study immunity in the course of complex treatment for malignant gliomas of the brain and to evaluate extracorporeal antigen-specific immunotherapy (EASIT), a pilot procedure which was carried out according to an approved protocol. Initially, lowered HLA-DR+ monocyte count and in vitro inhibition of proliferative activity were reported in all patients. Inductive EASIT started in early postoperative period aborted immune disturbances caused by surgery. In 1998-2000, the procedure was performed in 33 patients with anaplastic astrocytoma (AA) (20) and glioblastoma (GB) (13). Mean dose of cell infusion was 2.43(0.18 x 109/patient and was well tolerated. There are 22 survivors and 9 patients died (GB--4 and AA--5; overall mortality--29%). Mean relapse-free survival was 14.2 mo (22); stable remission during 12-18 mo--37.5% (3/8)(GB) and 64% (9/14) (AA) Complete rehabilitation of immunity was generally reported 12 mo after the course of EASIT. Hence, complex treatment (surgery + EASIT) enhanced its efficacy in the management of brain tumors.

[Characteristics and immunologic changes in patients with malignant brain tumors]. / Kharakteristika i mekhanizmy immunnykh narushenii u bol'nykh so zlokachestvennymi opukholiami golovnogo mozga.

The investigation was concerned with assaying immunity and evaluating the role played by monocytes and tumor cells in the formation of T-cell dysfunction in malignant glioma (MG). The study group included 28 patients with anaplastic astrocytomas (n = 18) and glioblastomas (n = 10). MG patients showed significant changes in the numbers of CD16+ NK-cells and HLA-DR monocytes as well as lowered levels of HLA-DR expression on monocytes and proliferative response of T-lymphocytes as compared with both standard and alternative pathways of activation. Monocytes and macrophages suppressed T-cell activity due to production of prostaglandins E2 in such patients. Enhanced immunosuppression was also reported in 24-hour supernatants of tumor cells. Immune disorders were shown to involve apoptosis-independent mechanisms. Hence, despite the immune privilege of the brain, immunocompetent cells crossed blood-brain barrier and counteracted with tumor cells. As a consequence, monocyte function and cellular cooperation dropped while production of immunosuppressive factors rose, and T-cell dysfunction was brought about through apoptosis-independent mechanisms.

[Efficiency of the use of peppermint (Mentha piperita L) essential oil inhalations in the combined multi-drug therapy for pulmonary tuberculosis]. / Effektivnost' ispol'zovaniia ingaliatsiia éfirnogo masla miaty perechnoi legkikh.

The essential oil of peppermint (Mentha piperita L.) has been found to have an in vitro pronounced and equal antimycobacterial effect in doses of 300 and 600 micrograms/ml, respectively. The use of its inhalations (upon 20-min heat evaporation into the room atmosphere for 2 months) as a supplement to combined multidrug therapy for pulmonary tuberculosis has indicated their significantly high positive effect in terms of abacillation (by 26.8 and 58.5% with doses of 0.01 and 0.005 ml/m3, respectively). This was followed by earlier positive X-ray changes in the lung and by attenuation of the intoxication syndrome. The findings suggest that peppermint essential oil may be used in combined multidrug therapy in patients with disseminated and infiltrative pulmonary tuberculosis.

[T cell subsets undergoing apoptosis and anergy in patients with pulmonary tuberculosis]. / Subpopuliatsionnaia prinadlezhnost' T-kletok, podverzhennykh anergii i apoptozu u bol'nykh tuberkulezom legkikh.

T-cell apoptosis and anergy as possible causes of impaired antigen specific responses and their subpopulation targets in patients with pulmonary tuberculosis were investigated. A decrease in PPD-stimulated proliferative responses were revealed in 43% of the examinees. The impaired PPD response was shown to be associated with both increased lymphocytic apoptosis and the arrest of cell cycle progression. CD4 and CHD8 T cells underwent apoptosis in PPD-stimulated cultures. Whereas a moderate apoptosis of CD4 cell could occur in PPD-reactive patients, accelerated apoptosis of CD8 cells developed only in PPD-unresponsive group. Both T-cell subpopulations displayed a decreased count of cells in S,G2/M phases of a cell cycle. Similar to apoptosis, the anergy of CD8 T cells was typical of PPD-unresponsive patients. Elevated apoptosis and anergy of CD4 and CD8 T cells in vitro were accompanied by a decline in the proportion of T cells and their subpopulations in patients with impaired PPD responses.

[Angiogenin and its role in angiogenesis]. / Angiogenin i ego rol' v angiogeneze.

The review is devoted to angiogenin, one of the factors that induce formation of blood vessels, which is unique among them in that it is a ribonuclease. Consideration is given to the tertiary structure of human angiogenin; the catalytic and cell-receptor binding sites, their significance for angiogenic activity; the human angiogenin gene structure, chromosomal localization, and expression; the specificity of angiogenin as a ribonuclease and abolishment of protein synthesis; the nuclear localization of angiogenin in proliferating endothelial cells and its significance for angiogenic activity; angiogenin binding to a cell-surface actin as a plausible mechanism of inducing neovascularization (enhancement of plasminogen activation by actin with angiogenin, stimulation of the cell-associated proteolytic activity by angiogenin; promotion of the cultured cells invasiveness); modulation of mitogenic stimuli in endothelial, smooth muscle, and fibroblast cells by angiogenin. The importance of angiogenin as an adhesive molecule for endothelial and tumor cells is discussed too, as well as the modulation of tubular morphogenesis by bovine angiogenin, prevention of tumor growth in vivo by angiogenin antagonists, prospects of the use of angiogenin and angiogenin-encoding recombinant plasmids and vaccinia virus in therapeutic practice.

[Involvement of nitric oxide in the development of tuberculin anergy in patients with pulmonary tuberculosis]. / Uchastie oksida azota v razvitii tuberkulinovoi anergii u bol'nykh tuberkulezom legkikh.

The production of nitric oxide (NO) and the magnitude of an antigen-specific proliferative response of the human lymphocytes stimulated by M. tuberculosis antigen [a purified protein derivative (PPD)] were investigated. PPD-reactive T lymphocytes were found in the peripheral blood of healthy donors. Normal values (mean values, the range of the minimum and maximum values) of PPD-induced proliferation and NO production were determined. Patients with pulmonary tuberculosis were found to have different levels of PPD-stimulated proliferation and NO production. The lymphocytes are shown to preserve their PPD reactivity in patients with normal NO production whereas the PPD-induced proliferative response was significantly decreased in those with high NO production. Patients with reduced tuberculin reactivities and high NO production were less responsive to treatment. The findings suggest that nitric oxide is involved in the development of tuberculin anergy with pulmonary tuberculosis.

[Immunomodulating effect of locoregional cytokine therapy in patients with pulmonary tuberculosis]. / Immunokorrigiruiushchii éffekt lokoregional'noi tsitokinoterapii u bol'nykh tuberkulezom legkikh.

The immunomodulating effect of local and regional cytokine therapy was studied in patients with different forms of pulmonary tuberculosis. The signs of immunosuppression either preserve or progress with tuberculostatic chemotherapy (a control group). The inclusion of cytokine therapy into a treatment regimen for patients with pulmonary tuberculosis (an experimental group) assures complete correction of immune disorders in 30% of patients, as manifested by a significant increase in the absolute count of T lymphocytes, in the relative content of CD8 cells and monocytes with HLA-DR antigen expression. The most pronounced effect was achieved in patients with a fibrocavernous form of pulmonary tuberculosis. Immunological correction was accompanied by positive clinical and laboratory changes. Arrested intoxication, improved X-ray pattern of the lung, and ceased bacterial isolation were recorded in the patients. Positive clinical and immunological changes suggest that it is expedient to include local and regional cytokine therapy as part of treatment in patients with pulmonary tuberculosis.

[Specific features of immunity in patients with different forms of pulmonary tuberculosis]. / Osobennosti immuniteta u bol'nykh s razlichnymi formami tuberkuleza legkikh.

The cellular immunity was studied in 59 patients with pulmonary tuberculosis. The development of tuberculous infection was ascertained to be accompanied by decreases in the relative counts of CD9, CD8, and CD72 lymphocytes, as well as monocytes, expressing class II histocompatibility antigens (DR). The patients with tuberculosis were found to have suppressed proliferative T-cell activity and IL-2 production, moderately decreased IL-1 production and increased TNF alpha secretion.

[Secondary immunodeficiency of medical staff contacting with tuberculous and nonspecific infection]. / Vtorichnyi immunodefitsit meditsinskogo personala, kontaktiruiushchego s tuberkuleznoi i nespetsificheskoi infektsiei.

The present paper analyzes immunological parameters and the incidence of secondary immunodeficiency (SID) in physicians and medium-levelled medical staff contacting with tuberculous and non-specific infection. Suppressed cell immunity was recorded in 44% of the medical staff of a pulmonary surgical tuberculosis hospital, with increased length of service there was a rise in the number of patients diagnosed as having immunodepression. The clinical manifestations of SID were recorded in 56% in this group and they were most pronounced in a group of long-working personnel. The proportion of persons with immunodepression proved to be twice higher among nurses than among physicians. Nurses are at the highest risk for immunopathological states. This common occurrence of SID among medical staff is an indicator to make an obligatory regular immunological examinations of the staff for the prevention and immunotherapy of SID.

[Use of xenogeneic spleen perfusate in the complex treatment of patients with destructive pulmonary tuberculosis]. / Primenenie perfuzata ksenoselezenki v kompleksnom lechenii bol'nykh destruktivnym tuberkulezom legkikh.

The first experience gained in the use of xenospleen perfusate in the management of 13 patients with disseminated fibrous-cavernous tuberculosis is presented. The disease was accompanied by an evident intoxication and respiratory insufficiency. Traditional ways of treatment failed to be effective in all the patients, thus indicating an intravenous drop by drop administration of xenogeneic spleen perfusate in the dose of 10 ml per kg body weight. The antiinflammatory (immunomodulation) effect manifested itself in the body temperature drop, less intoxication and infiltration processes in the lungs, as well as a better hemogram.