IFN-α, IFN-ß, and IFN-γ Have Different Effect on the Production of Proinflammatory Factors Deposited in Weibel-Palade Bodies of Endothelial Cells Infected with Herpes Simplex Virus Type 1.

We demonstrated similarities and differences in the effects of IFN-α and IFN-ß compared to IFN-γ on the production of factors deposited in the Weibel-Palade bodies in cultures of endothelial cells (intact and infected with herpes simplex virus 1). IFN-α and IFN-ß reduced the content of von Willebrand factor, endothelin-1, and soluble P-selectin and increased IL-8 concentration in the culture medium of human umbilical vein endothelial cells. IFN-γ reduced the content of all studied factors in the endothelial cell culture medium. Possible mechanisms of these effects are discussed.

Effects of gold nanoparticles on erythrocyte hemolysis.

We studied hemolytic activity of gold nanoparticles added to the whole blood (ex vivo) and of nanoparticles coated and not coated with plasma components on erythrocytes in hypotonic medium (osmotic hemolysis) in vitro. Gold nanoparticles did not stimulate erythrocyte hemolysis after 4-h incubation with the whole blood ex vivo. Hemolysis tended to increase in the presence of small gold nanoparticles (5, 10, 20 nm) at the maximum concentration of 20 µM (by gold content) used in our study in comparison with the control. This tendency was detected during the 1st hour of the nanoparticles incubation with blood. Gold nanoparticles in the used concentrations (up to 20 µM of gold) coated with plasma components after preincubation with autologous plasma and nanoparticles without coating caused no osmotic hemolysis of erythrocytes in vitro.

[Human interferon modulates infected vascular endothelium function].

BACKGROUND: To study impact of interferon (IFN) alpha, beta and gamma on the Herpes simplex virus type 1 (HSV-1) infected endothelial cells functional activity related with participation in the inflammation development. MATERIALS AND METHODS: In the work endothelial cells isolated from umbilical vein were used. Intact and infected cultures were treated by interferon and in the dynamics of cultivation tested mediators in the cultural medium. RESULTS: All investigated interferons activated the production of IL-6. IFN alpha, beta activated the production of IL-8, while IFN gamma inhibited her. IFN alpha and gamma increased synthesis of nitrogen oxides and reduced the synthesis of endothelin-1, while IFN beta activated the production of endothelin-1. CONCLUSION: Infection of endothelial cells isolated from umbilical vein with HSV-1 does not alter the ability of interferon in modulating of proinflammatory cytokines, nitric oxide and endothelin-1 synthesis. It is obvious in the body modulation manifestations of innate immunity under the influence of exogenous interferon is implemented both intact and infected with HSV-1-vascular endothelium and nature modulation is determined by the type of IFN.

[Changing the protective properties of the receptor domain of protective antigen anthrax exotoxin, depending on the orientation of the presentation on nanoparticles].

Polysterene particles bearing on its surface recombinant protein receptor domain of protective antigen anthrax exotoxin, exposed in different orientations have been constructed. Particles with exposed COOH region of receptor domain induced the highest protective immunity in mice anthrax model (95%). We revealed that immunization with these particles causes a specific induction of Th1-response, characterized by increased levels ofcytokine TNF-alpha and IL-2.

[Differences in functional activity of cultivated human vascular endothelium received from different donors].

Endothelium of blood vessels in the organism is involved in carrying out numerous functions in normal and pathological processes. Development of the method of isolation and cultivation of endothelial cells has made it possible to model the processes occurring in vascular endothelium. Unlike continuous cell lines, research on primary cell cultures lead to wide variation in results. In this study, spontaneous production of markers characterizing functional activities of endothelium were compared in endothelium cultures derived from umbilical cords of 20 donors. It was found that, based on the production levels of all investigated markers after 3 hours of cell cultivation, these cultures can be divided into high- and low-producing. Analysis of cytokine profiles revealed that the level of spontaneous production of IL-1beta in these groups did not vary during cell cultivation up to 24 and 48 hours, whereas the levels of IL-6 and IL-8 production increased to 24 and 48 hours, and the difference between groups became leveled; the increase in production of TNFalpha occurred only in cultures of low-producing group. The increase in amounts of sP- and sE-selectin in cultural medium was observed only under cultivation of low-producing cultures, whereas the increase in sICAM-1 was noted under cultivation of highly-producing cultures; the increase of sPECAM-1 was revealed under cultivation of both highly- and low-producing cultures. So, the difference in the levels of this CAM between the groups remained. The levels of sVE-cadherin in cultural medium did not vary in the course of cell cultivation. The levels of nitrite reflecting the amount of NO were increased in cultural medium in all cultures, and the difference between the groups remained; concentration of endotelin-1 was increased, however the values of this marker in the cultural medium of several cultures were similar, therefore, it was not possible to create groups reflecting levels of its production. The levels of von Willebrand Factor were increased in cultural medium under cultivation of cultures of both groups, however the difference between the groups did not remain. The levels of matrix metalloproteinase-1 in cultural medium increased under cultivation of cell cultures. Hence, endothelial cultures from different donors differ in their ability to produce markers of functional activity, and reflect the features of cell donors. The results obtained allow modeling the processes occurring in vascular endothelium taking into account the individual characteristics of cultures, and suggest the possibility of a more thorough approach to evaluating the results obtained using primary endothelium cultures.

[Adhesion molecules expressed in vascular endothelial cells in natural immunity against viral infections].

Cell adhesion molecules (CAM) expressed in vascular endothelium ensure integrity of the endothelial layer, recruitment and transmigration of leukocytes. Being receptors of many viruses, they play a role in immune control and infectious processes. Monoclonal anti-ICAM-1 antibodies enhance infection of primary human umbilical vein endothelial cell (HUVEC) cultures with HIV-1 due to incorporation into virions. IFN-gamma activates expression of ICAM-1 on HIV-infected HUVEC and thereby promotes binding of this molecule to complementary molecules on a greater number of sensitive cells, virion transfer onto them, and broad dissemination of the virus. Recombinant human IFN-alpha, IFN-beta and IFN-gamma influence (activate, inhibit) CAM shedding from HUVEC both intact and infected wit HSV-1. Activated shedding in the blood stream due to competition between soluble and endothelial CAM slows down recruitment and transmigration of leukocytes, i.e. regulates inflammation. CAM incorporated in microparticles can influence a wide spectrum of pathological processes Endothelial CAM may be a target for the delivery of pharmaceuticals for the treatment of vascular (including infectious) pathology.

Lipophilic Prodrug of Methotrexate in the Membrane of Liposomes Promotes Their Uptake by Human Blood Phagocytes.

Previously, we showed that incorporation of methotrexate (MTX) in the form of a lipophilic prodrug (MTXDG) in 100-nm lipid bilayer liposomes of egg phosphatidylcholine can allow one to reduce toxicity and improve the antitumor efficiency of MTX in a mouse model of T-cell leukemic lymphoma. However, in our hemocompatibility tests in vitro, MTX liposomes caused complement (C) activation, obviously due to binding on the liposome surface and fragmentation of the C3 complement factor. In this work, we studied the interactions of MTX liposomes carrying stabilizing molecules phosphatidylinositol (PI), ganglioside GM1, or a lipid conjugate of N-carboxymethylated oligoglycine (CMG) in the bilayer with subpopulations of human blood leukocytes. Liposomes labeled with BODIPY-phosphatidylcholine were incubated with whole blood (30 min and 1 h, 37°C), blood cells were lysed with a hypotonic buffer, and the fluorescence of the liposomes bound but not internalized by the leukocytes was quenched by crystal violet. Cell suspensions were analyzed by flow cytometry. Incorporation of MTXDG dramatically enhanced the phagocytosis of liposomes of any composition by monocytes. Neutrophils consumed much less of the liposomes. Lymphocytes did not accumulate liposomes. The introduction of PI into MTX liposomes practically did not affect the specific consumption of liposomes by monocytes, while CMG was likely to increase the consumption rate regardless of the presence of MTXDG. The GM1 ganglioside presumably shielded MTX liposomes from phagocytosis by one of the monocyte populations and increased the efficiency of monocyte uptake by another population, probably one expressing C3b-binding receptors (C3b was detected on liposomes after incubation with blood plasma). MTX liposomes were shown to have different effects on TNF-α production by activated leukocytes, depending on the structure of the stabilizing molecule.

[Angiogenesis and viral infections].

The review summarizes the results obtained from a study of the mechanisms responsible for angiogenesis during embryogenesis and adulthood in various abnormalities and in infections caused by hepatitis C virus and herpes simplex virus.

[Human blood vascular endothelial dysfunction in HIV infection].

The review presents information on the impact of human immunodeficiency virus (HIV) on the coagulative system in blood vessels and on the potential of endothelial infection in the organism and in the culture of endothelial cells derived in vitro from blood vessels of different tissues of the organism. It also discusses information on the mechanisms responsible for angiogenesis and apoptosis of the endothelial cells cultured in vitro under the influence of HIV and its proteins.

[Effect of staphylococcal enterotoxin A on the development of Lewis lung carcinoma in mice]. / Vliianie stafilokokkovogo énterotoksina A na razvitie kartsinomy legkogo L'iuis u myshei.

The effect of Staphylococcus aureus enterotoxin A (SEA) was studied for its effect on the development of the Lewis carcinoma in mice. It was shown that administration of SEA immediately after the appearance of the primary node in mice after transplantation of tumour cells led to insignificant inhibition of the node growth and a slight decrease of tumour metastasizing into the lungs. Inoculation of mice after the appearance of the primary node with 1/microgram of SEA 5 times a week significantly increased their survival rate. The lack of the marked effect of SEA appears to be associated with the disturbance of the immune interferon system functioning in tumour-bearing mice, since the production of serum interferon induced by SEA in mice with tumours was considerably lower than in the intact ones.

[Effect of splenic cells sensitized with staphylococcal enterotoxin A on the metastasis of Lewis lung carcinoma in mice]. / Vliianie kletok selezenki, sensibilizirovannykh stafilokokkovym énterotoksinom A, na metastazirovanie kartsinomy legkikh L'iuis u myshei.

The effect of intact mouse spleen cells sensitized in vitro with staphylococcus aureus enterotoxin A (SEA) on spreading of mouse Lewis carcinoma was studied. A significant decrease in a number of metastases in the lungs and in the weight of the lungs was observed after multiple intrapulmonary inoculation of spleen cells treated with SEA for 6 hours. The effect was less marked after inoculation of the sensitized cells intraperitoneally or into the femoral muscle of the leg affected with the tumour. After multiple inoculations of the sensitized cells, the spleen cells of the treated animals develop the state of interferon hyporeactivity to SEA but not to PHA or NDV.

[Non-specific antitumoral activity of staphylococcal enterotoxins]. / Protivoopukholevaia nespetsificheskaia aktivnost' stafilokokkovykh énterotoksinov.

St. aureus enterotoxins A and B possess an antitumour effect. After intraperitoneal inoculation they decrease the size and in some cases prevent the development of the human hypernephroma in the cheek pouch of golden hamsters. The effect of enterotoxins may possibly consist in inducing the production of endogenous immune interferon which activates the host immune system and enhances the rejection of heterologous tumour cells.

The course of influrenza infection in mice with graft-versus-host reaction.

The course of influenza infection in mice with a developed graft-versus-host reaction (GVHR) was changed. Due to disturbances in the inflammatory process the pneumonia was delayed and less marked. Consequently, the infected mice died later than controls. Influenza virus reproduction in the lungs was more intensive and its persistence more prolonged. Interferon production in the lungs of mice with GVHR was similar to that in the controls.

Comparative study on the anticellular and antiviral effects of interferon and the haemopoietic stem cell inhibition factor.

Pretreatment with crude interferon preparations obtained from suspension cultures of bone marrow, spleen and thymus cells or from mouse L-cell cultures or with mouse serum interferon preparations did not change the colony-forming activity of bone marrow cells on syngeneic transplantation to lethally irradiated mice. Preparations of L-cell culture interferon, dialysed and purified by carboxymethyl-Sephadex (G-25) column chromatography, showed an inhibitory effect on exogenous colony formation by bone marrow cells. The results suggested the presence in crude interferon preparations of a substance either inhibiting the anticellular effect of interferon or stimulating colony formation. The factor produced by thymus cells following their treatment with antilymphocyte serum inhibited colony formation by bone marrow cells and, unlike interferon, possessed no antiviral activity when tested in cell cultures.

[Preparation and properties of the staphylococcal toxin causing the toxic shock syndrome]. / Poluchenie i nekotorye svoistva stafilokokkovogo toksina, vyzyvaiushchego sindrom toksicheskogo shoka.

A method is proposed for isolation and purification of the staphylococcal toxin causing the toxic shock syndrome (TSS). The method includes three steps: aggregation of protein from the cultural filtrate of Staphylococcus aureus strain 1169 in the presence of 0.025% sodium hexametaphosphate at pH 3.0; gel filtration of the concentrated material on the sephadex G75; ion-exchange chromatography on DEAE 32 cellulose. The proposed method permits to obtain the purified biologically active preparation of toxin with the yield about 40%. The obtained preparations are homogeneous in polyacrylamide electrophoresis and as analyzed by immunochemical methods. The mol mass of the isolated protein is 24 kD, it is not immunologically identical to staphylococcal toxins A-D and is lethal for New Zealand white rabbits and chinchilla rabbits. Interferon inducing activity of the protein is identical to the one of staphylococcal enterotoxin type A.

[New data on acid-labile alpha-interferon]. / Novoe o kislotlabil'nom alpha-interferone.

Reviews the information on the new concepts about the composition and properties of abnormal alpha-interferon-alpha-acid-labile interferon (alpha-ALI) detected in the blood of patients with AIDS, autoimmune, and other diseases. alpha-ALI includes alpha- and gamma-interferon which is responsible for acid lability. Increased content of serum alpha-ALI and its acid lability are poor prognostic signs in AIDS. A decrease in these values, concomitant with clinical improvement in some autoimmune diseases, indicates involvement of alpha-ALI in the pathogenesis of these diseases. Activating effect of gamma-interferon in vitro is demonstrated on different models, including HIV infection of human vascular endotheliocytes. The effect of alpha-ALI in patients is apparently determined by the ratio of its components alpha- and gamma-interferons.

[Spectrum of cytokines produced by blood leukocytes in infection of herpes simplex virus, type 1]. / Spektr tsitokinov, produtsiruemykh leikotsitami krovi pri infektsii virusom prostogo gerpesa tipa 1.

Three types of reaction of human blood leucocytes to herpes simplex virus 1 (HSV-1) were detected. The first reaction type, i.e. production of IFN-alpha, IFN-gamma, IL-1beta, IL-6 and TNFalpha but not of IL-2 or IL-4, denotes the primary body reaction to an infectious agent. The second reaction type is related with infection of activated HSV-1 leucocytes and is accompanied by an inhibited production of IFN-gamma, IL-6 and IL-8, which is targeted at suppressing the antivirus cell mechanisms. The third reaction type is associated with production of IFN-alpha, IFN-gamma, IL-beta, IL-6 and TNFalpha by blood leucocytes affected by HSV-1-infected leucocytes.

[Comparative study of interferon production in mice with the graft versus host reaction]. / Sravnitel'noe izuchenie produktsii interferona u myshei s reaktsiei transplantat protiv khoziaina.

Mice with graft versus host reaction (GVHR) show a decreased production of serum interferon and that produced by the bone marrow and spleen cells and blood leukocytes in vitro upon inoculation with Newcastle disease virus. Interferon induction with lipopolysaccharide of Flexner bacteria resulted in activation of production of serum interferon and that induced in spleen cell and blood leukocyte suspensions. Serum interferon production after administration of poly(I) . poly(C) was similar in mice with GVHR and controls.

[Interferons produced by human leukocytes after induction with influenza virus]. / Interferony, vyrabatyvaemye leikotsitami cheloveka posle induktsii virusom grippa.

Influenza virus induces in human leukocytes the production of 3 types of interferon: 1--acid-labile, produced up to 3 days; 2--acid-stable, produced for 24 hours; 3--acid-stable, produced up to 3 days and showing antiviral activity in heterologous culture of cow embryo lungs alone. The heterogeneity of properties of the 3 interferon types appears to reflect heterogeneity of molecular weights of its components.

[Feasibility of using human spleen cells to produce interferon]. / Vozmozhnost' ispol'zovaniia kletok selezenki liudei dlia produktsii interferona.

Spleen cells of subjects who died suddenly are capable of producing large amounts of interferon in response to induction with Newcastle disease virus (NDV) and Sendai virus (1-2.5 IU per 1000 cells). The optimal conditions for interferon production by these cells include using NDV, the H strain, as an inducer in a dose of 10 ID50/cell and fresh spleen cells stored no more than 24 hours at 4-6 degrees C in a concentration of 0.5-1.0 x 10(7)/ml, with the incubation time of 20-24 hours. It is recommended that spleen cells from subjects who died suddenly be used for human interferon production.