Human Interferon Preparations Research Articles

CURRENT ISSUES OF ANTIVIRAL THERAPY OF ARVI AND INFLUENZA IN MILITARY UNITS

Conducting preventive medical and epidemiological measures for influenza and ARVI in closed military units, including on ships, vessels and auxiliary units of the Navy, is a factor of increasing the combat effectiveness of forces of the Russian Army. In the structure of total morbidity of military personnel, acute respiratory viral infections account for 35–50%, however, during the young military reinforcement, the incidence of these infections in combination with community-acquired pneumonia can reach 70–80%. The risk of exceeding the epidemiological threshold for respiratory tract infections, the etiological basis of which is viral aggression, is especially important in newly formed military contingents, for units participating in local military conflicts and peacekeeping operations, as well as with a new recruitment of cadets at training centers. The use of non-specific antiviral drugs based on exogenous recombinant interferon -2b is one of the conditions for adequate prevention of influenza and ARVI in closed military units. The paper presents the results from the analysis of publications on the effectiveness and Viferon® safety in the treatment of influenza and ARVI. Viferon is a preparation of recombinant human interferon -2b with a complex of antioxidants in various dosage forms, used in the treatment and prevention of acute respiratory viral infections and influenza. Use of Viferon in various dosage forms is a reliable method of prevention and treatment of influenza and other acute respiratory viral infections in military forces, including on ships, vessels and auxiliary units of the Navy, at risk of emergency epidemiological situations due to ARVI and influenza, in conditions of limited contingent.

Значення наночастинок у транспорті інтерферону

The article analyzes the materials of scientific works on the preparation and use of preparations containing interferon and nanoparticles of metals or proteins. The application of a new innovative nanotechnology method using nanoparticles provides broad opportunities for improving the targeted delivery of drugs to the pathological process, implementing an optimal toxicity profile. The purpose of our work was to conduct an analysis of scientific works on the introduction into the medical practice of interferon complexes with nanoparticles of metals or proteins, to identify their advantage and provide a characteristic. It has been established that the nanocomplex obtained can enhance the effectiveness of interferon, reduce its side effects and have controlled release of the drug in the body. Among the antiviral nano-drugs, are actively involved in medical practice - interferon and its derivatives. The system of interferon is on the first line of protection of the organism from infection. Interferon acts on all viruses that enter the body. Taking into account the viral nature of some types of cancer, the interest in using IFN drugs in oncology has increased significantly after their high antiviral activity has been shown. Positive results have been found in the use of IFN in the treatment of chronic hepatitis B and C, condylomatosis and several tumor-like neoplasms, for example, AIDS-related measles, chronic myeloid leukemia, follicular lymphoma, T-cell lymphoma, carcinoma of the tumor, melanoma, and numerous renal cell carcinomas. Employees of the DNU ("State Center for Innovative Biotechnologies") are developing a new preparation of recombinant human interferon with low-dose cerium dioxide. The main purpose relied on this drug - the fight against viral infection and prolonged antitumor action. The named complex successfully runs preclinical and clinical studies. According to the results of experiments in vivo, scientists were able to prolong the action of interferon ox 2 to 5 days. The anti-herpetic activity of cerium dioxide nanoparticles on the model of genital herpes in cavy has been investigated. Numerous experiments indicated the prophylactic and therapeutic efficacy of the use of a complex of cerium dioxide and interferon nanoparticles in a concentration of 1.25 mM In the future, the continuation of work on the creation of a new pharmaceutical drug based on interferon and nanoparticles of the metal to optimize its effectiveness and reduce side effects.

A new contained human immunodeficiency virus type 1 host cell system for evaluation of antiviral activities of interferons and other agents in vitro

HIV-host infection systems in vitro are important in the pre-clinical assessment of anti-retroviral drug activity. The present report describes the development of a new HIV-host model comprised of an epithelial cell line of HeLa lineage (HeLa-1), transfected with expression vectors bearing tat and rev (TART) genes of HIV-1 as well as the CD4 receptor gene, and HIV-1 ΔTat/Rev, a biologically contained strain of HIV-1 deleted in tat and rev. Measurement of infectivity, by syncytium formation and reverse transcriptase assay, revealed that HeLa-1 is infected with HIV-1 ΔTat/Rev. This virus failed to productively infect the TART-deficient CD4-positive HeLa cells, confirming its contained, non-infectious nature. The HeLa-1/HIV-1 ΔTat/Rev system was used to measure the anti-retroviral activity of a human leukocyte-derived interferon (IFN-αn3) preparation, several nucleoside analogs, and protease inhibitors. The HeLa-1/HIV-1 ΔTat/Rev model provides a biologically contained system for the study of the HIV pathogenesis and the relative and combined therapeutic effects of anti-retroviral agents in vitro.

Insusceptibility of Cockayne syndrome-derived lymphocytes to plasminogen activator-like protease induction by ultraviolet rays and its abolition by interferon

Protease induced by ultraviolet rays (UV) has been extensively investigated in human cells. Plasminogen activator-like protease (PA) activity was induced soon after UV irradiation in peripheral lymphocytes derived from healthy donors. In contrast, UV-irradiated lymphocytes derived from Cockayne syndrome (CS) cases did not show marked protease inducibility. Epstein-Barr (EB) virus-transformed CS lymphoblastoid cells were also characterized by insusceptibility to UV-induction of PA. However, when CS-derived cells were treated with a human interferon (HuIFN) preparation prior to irradiation, induction of PA activity was detected, irrespective of the kind of HuIFN (α or γ). The results indicate the possibility of abnormal PA metabolism in CS-derived cells.

Enhancement of an anti‐tumor effect of interferon by dipyridamole in established human malignant melanoma cell lines

Enhancement of the anti-proliferative effect of human interferon (HuIFN) preparations (alpha, beta and gamma) by dipyridamole was detected in a human malignant melanoma cell line, MM-ICB, which we originally established. Cell growth was inhibited by HuIFN alone, but a marked increase in inhibition was noted in vitro and in vivo when dipyrydamole was added. Cellular DNA synthesis, as determined by 3H-deoxythymidine incorporation into the acid-insoluble cellular fraction, was more inhibited by combined treatment than by any of the agents used alone. Two other melanoma cell lines that we established, MM-2CB and MM-3CB, also exhibited sensitivity to combined treatment both in vitro and in vivo. Furthermore, the HMV-I and SEKI melanoma cell lines were susceptible to the combination. Even non-cytotoxic concentrations of dipyridamole could enhance the effect of HuIFN on MM-ICB, MM-2CB, and SEKI cells.

Establishment and characterization of a human cell strain, KT, with high sensitivity to UV-killing and to cell proliferation inhibition by interferon.

We have established a human cell line, designated KT, with high susceptibility to both cell proliferation inhibition by interferon and UV‐killing, from a metastatic breast carcinoma. A tumor marker, a pregnancy‐specific glycoprotein (Schwangerschaftsprotein 1; SP1), and carcinoma characteristics compatible with ductal carcinoma of the breast were seen in KT cells by electron microscopic observation. KT cells were slightly more resistant to X‐ray‐induced toxicity than fibroblastic cells, termed KS, from the scalp of the patient. But, KT cells had lower cloning efficiency after UV irradiation than did KS cells: D0 values of 1.5 J/m2 and 7.2 J/m2, respectively. KT cells also appeared more susceptible to human interferon (HuIFN) preparations (α, β, γ and natural or recombinant) than did KS cells, as measured by cell colony formation ability, proliferation rates, and [3H]deoxythymidine incorporation levels into acid‐insoluble cell materials. The sensitivity of KT cells to UV and HuIFN was greater than that of human RSa cells, a cell line with high sensitivity to both agents. KT cells had more capacity for UV‐induced DNA‐repair replication synthesis than did RSa cells, the capacity being much the same as that of KS cells. There was no significant difference in levels of antiviral activity induced by HuIFN and binding capacity for 125I‐labeled IFN‐αA between KT and KS cells. KT cells appeared refractory to cell proliferation inhibition by tumor necrosis factor (TNF) preparations.

Effects of human alpha-interferon on granulocyte-macrophage progenitor cells (CFU-GM) in vitro.

Two preparations of human interferon (IFN)-alpha were assessed for their influence on granulocyte-macrophage progenitor cells (CFU-GM) in vitro. Both highly purified human IFN-alpha Ly and recombinant IFN-alpha 2a suppressed CFU-GM colony formation in a dose-dependent manner using low-density bone-marrow target cells. Suppression of CFU-GM colony formation was accompanied by an increase in clusters. However, depletion of monocytes, T lymphocytes and B lymphocytes from low-density bone-marrow cells resulted in insensitivity of progenitor cells to IFN-alpha. These results demonstrate that the effects of human IFN-alpha on myeloid progenitor cells (CFU-GM) are mediated by accessory cells within the bone marrow.

Effects of human interferon on cellular response to UV in UV-sensitive human cell strains

Cells of human RSA cell line, with high senstivity to UV killing and low capacity for DNA repair, when pretreated with 1–100 units/ml of human interferon (HuIFN) preparations for more than 12 h before irradiation, acquired an enhancement of UV-induced DNA-repair relications synthesis in association with recovery from inhibition of total cellular DNA synthesis and UV survival. Prompt and transient induction of plasminogen activator activities was also found within 5 min after UV irradiation in the cells pretreated with HuIFN but not in the cells non-pretreated with HuIFN. The enhancement and induction effects of HuIFN were observed, irrespective of the kind of HuIFN preparation used (α, β or γ, and natural or recombinant) and in other UV-sensitive fibroblast cells which were derived from Cockayne syndrome and xeroderma pigmentosum fibroblasts (XP1KY). However, all of the enhancement of DNA-repair synthesis and the induction of plasminogen activator activities by HuIFN was suppressed by treatment with cycloheximide immediately after UV irradiation.

Comparative effects of α, β and γ human interferons on a HBsAg-producing human hepatoma cell line

The effects of all the three types of human interferons (alpha, beta and gamma) on a human hepatoma cell line with hepatitis B virus (HBV)-DNA sequences integrated into the host DNA and producing hepatitis B surface antigen (HBsAg) in culture medium were assayed. The aim of the present research was to test human interferon preparations in an in vitro system for hepatitis B virus, and to compare the observed effects. The results evidenced both the antireplicative activity principally showed by preparations of beta and gamma human interferons and the inhibition of HBsAg production by high concentrations of gamma human interferon.

15] Preparation and partial purification of human interferon 8

Publisher Summary This chapter discusses the preparation and partial purification of human interferon δ. The most surprising characteristic of this interferon is that it appears to induce the antiviral state only in cells trisomic for chromosome 21, which are more sensitive than diploid cells to the classical interferons. This suggests that the major role of this interferon may be other than the protection of cells against virus infection and that its antiviral effect is indeed incidental to its main (unknown) function. In this respect, it is worth noting that macrophage-activating factor (MAF) has been shown to be identical with IFN-γ, and that indeed this protein is more potent in activating macrophages than as an antiviral agent. Study of IFN-δ is not aimed at total purification, but instead the objective is to separate IFN-δ from IFN-γ to characterize it. The chapter describes simple techniques for achieving this separation. A novel interferon type, present in supernatants from human peripheral blood mononuclear leukocytes (PBML) induced with phytohemagglutinin and teleocidin, may be easily separated from the main component (IFN-γ) by simple chromatographic procedures. The activity is easily detected on human cells trisomic for chromosome 21 but not on other cell lines commonly used for the assay of interferon.

11] Large-scale production and recovery of human immune interferon

Publisher Summary This chapter discusses the development of a method for large-scale production of human immune interferon based on induction scheme with the addition of a cell fractionation process and a novel method for product recovery. A 10- to 15-fold enhancement in interferon yield and a 50-fold increase in purity of crude product were accomplished by nylon wool column fractionation of lymphocytes combined with simultaneous reduction of autologous plasma protein levels prior to induction. The production method described is suitable for large-scale preparation of natural human immune interferon. Its principal advantages include greater interferon yield and purity without substantial increase in process costs. The process provides a method for the production of both higher and more consistent yields of interferon. A major factor in the higher yield from nylon wool fractionated buffy coat cells is the prolongation of the active production period compared to unfractionated cell preparations. Higher purity without concomitant yield loss results from removal of about 70% of buffy coat plasma, a step which results in reduced yields when using unfractionated cells. This sequential application of nylon wool fractionation and protein removal steps combines to provide 50-fold improvement in initial purity, which leads to higher purification yields and a reduction in purification costs.

89] Radioimmunoassay for detection of changes in cell surface tumor antigen expression induced by interferon

This chapter describes radioimmunoassay for the detection of changes in cell surface tumor antigen expression induced by interferon. The interferons are multifaceted biological modifiers that exert antiviral, antiproliferative, and several different immunomodulatory actions. Their ability to increase the amount of tumor antigens on the surface of human carcinoma cells would have potential utility in enhancing the in vivo detection and/or therapy of tumors with monoclonal antibodies in clinical trials. These antigens are recognized by a panel of monoclonal antibodies that are generated in laboratory with membrane-enriched fractions from liver metastases from patients with primary breast carcinoma. Partially purified preparations of human interferon have been shown to increase the amount of HLA-A,B,C and β 2 -microglobulin on the surface of several different human cell types. A radioimmunoassay with live cells is used to determine whether human recombinant leukocyte (alpha) A interferon (IFN-αA) could increase the presentation of tumor antigens on the surface of human breast and colon carcinomas as detected by monoclonal antibodies. The results suggest that recombinant human leukocyte interferon can increase the expression of tumor antigens on the surface of human breast and colon carcinoma cells.

Results of space experiment program “interferon”

The results of the biological space experiment “Interferon” performed by two international cosmonaut teams (26 May 1980, and 16 May 1981) aboard space laboratory Solyut-6 are reported: (1) Human lymphocytes separated from blood of healthy donors and placed into “Interferon I” equipment could be kept for 7 days in suspension culture under spaceflight conditons. Interferon production could be induced in human lymphocytes by preparations of different origin: virus, synthetic polyribonucleotides, bacterial protein and plant pigment. An increased lymphocyte interferon production in space laboratory compared to ground control was observed. (2) Human interferon preparations and interferon inducers placed in space laboratory at room temperature for 7 days maintained their biological activity. (3) A decrease of induced interferon production and natural killer activity of lymphocytes isolated from peripheral blood of cosmonauts was observed on the 1st day on Earth after 7-days spaceflight.

Effects of human interferon preparations on neutrophil function.

We have studied effects of two partially purified human leukocyte (alpha) interferon (IFN) preparations (PIF-A and PIF-B) and a highly purified fibroblast (beta) IFN on the functional activity of normal human neutrophils (PMNs). In vitro, PIF-B conferred a significant and dose-dependent enhancement of chemiluminescence (CL) induced both by phagocytosis and a soluble stimulus, f-Met-Leu-Phe, and decreased killing of Staph. aureus. In contrast, PIF-A caused only a slight inhibition of bactericidal activity and had no effects on CL. beta-IFN had no effects on either bactericidal activity or CL. Migration under agarose was decreased with all of the IFN but phagocytosis and release of enzymes was not affected. PMNs from seven patients treated with PIF-A for multiple myeloma exhibited increased CL responses but no other PMN functions were affected. The findings that human IFN preparations affect PMN functions indicate that high-dose IFN therapy of immunocompromised patients should be carefully evaluated for the possibility of increased infectious complications.

Studies on safety problem by skin tests using human beta interferon preparation.

Skin reactivities to human fibroblast-derived interferon (HuIFN-beta) preparation were studied in 20 healthy subjects in order to establish a method that detects hypersensitivity to the preparation. In response to intracutaneous injection of 0.02 ml of a 3 X 10(6) international reference units (IU)/ml of HuIFN-beta, the subjects developed immediate wheal-and-erythema type cutaneous reactions followed by erythemas maximum in diameter at 12 and 48 h. The immediate reactions were macroscopically similar to those observed in IgE-mediated cutaneous allergic reactions were macroscopically similar to those observed in IgE-mediated cutaneous allergic reactions elicited by intradermal injection of allergens. Little or no such reactions occurred with test solutions in concentration of less than 2 X 10(5) IU/ml and with solutions containing proteins of bovine origin, possible contaminants of the proteins in the preparation. These results suggest that the immediate cutaneous response to the solution of 3 X 10(6) IU/ml of HuIFN-beta is an essentially nonspecific reaction to HuIFN-beta or substances intimately associated with HuIFN-beta. For prediction of IgE-mediated hypersensitivity, a skin test with a solution containing HuIFN-beta in concentration of less than 2 X 10(5) IU/ml seems to be applicable. A test with a solution containing neonatal calf serum in concentration of 10-100 micrograms/ml may also be necessary as well as with the HuIFN-beta preparation.

Evaluation of absorption with Limulus amebocyte lysate to remove contaminating endotoxin from interferon and lymphokine preparations

α and β human interferon (IFN) preparations and lymphokines (supernatants of PHA-stimulated blood lymphocytes) were deliberately contaminated with endotoxin (20 ng/ml) and subsequently rendered endotoxin-free by absorption with Limulus amebocyte lysate (LAL). Absorption with LAL did not appreciably affect the antiviral activity of IFN and lymphokines in 8 experiments and caused a 30–50% reduction in two. The capacity of these agents to stimulate natural killer cell activity and monocyte cytotoxicity was not consistently modified by absorption on LAL. When the chemotactic activity of lymphokine for monocytes was measured, the maximal number of monocytes induced to migrate and the maximal active lymphokine concentration were not affected by absorption with LAL. LAL-treated lymphokines, however, showed a prozone phenomenon, presumably related to the release of chemotaxis inhibitor(s) from the LAL gel.

Determination of human interferons α, β and γ in plasma and/or pharmaceuticals

The fact that human Interferon is inactivated with protein as such as trypsin and pepsin has been noted, and a new method for quantitative determination was established. It was revealed that the decomposing substance of the human Interferon obtained by an action upon kallikrein or trypsin, under fixed conditions, hydrolyzed a substrate peptide MCA. By this method, a sensitive new method for quantitative determination of favorable reproducibility in human Interferon preparations and in human plasma was established.

Comparative analysis of the influences of human gamma, alpha and beta interferons on human multipotential (CFU-GEMM), erythroid (BFU-E) and granulocyte-macrophage (CFU-GM) progenitor cells.

Preparations of human interferon (HuIFN) immune (gamma) (2 X 10(7) units/mg protein), HuIFN leukocyte (alpha) (1.4 X 10(8) units/mg protein) and HuIFN fibroblasts (beta) (10(6) U/mg protein) were assessed for their influence on colony formation of human hematopoietic progenitor cells: colony forming unit-granulocyte, erythroid, macrophage, megakaryocyte (CFU-GEMM), burst forming unit-erythroid (BFU-E), day 7 colony forming unit granulocyte-macrophage (CFU-GM) and day 14 CFU-GM. Colony formation by CFU-GEMM and BFU-E was suppressed equally by the three preparations of HuIFN, but colony formation by CFU-GM was suppressed differentially. CFU-GM were, on the whole, more responsive to HuIFN gamma than HuIFN alpha, and HuIFN beta was least effective. HuIFN alpha, but not HuIFN gamma or HuIFN beta, suppressed colony formation from CFU-GM without also suppressing the total number of colonies plus clusters. This was due to an increase in the numbers of clusters formed in the presence of HuIFN alpha. The suppressive influence on colonies from CFU-GM by the preparations of HuIFN and the enhancement of clusters by HuIFN alpha was apparently equal for colonies and clusters of neutrophils, eosinophils, macrophages and neutrophils plus macrophages. The suppressive effects of HuIFN gamma were inactivated by a monoclonal antibody to HuIFN gamma and the suppressive and enhancing effects of HuIFN alpha were inactivated with a heteroantiserum to HuIFN alpha. Depletion of monocytes, T lymphocytes and B lymphocytes from the target bone marrow cells had no influence on the effects of the preparations of HuIFN. These results demonstrate that the effects of HuIFN gamma and HuIFN alpha are due to the HuIFN themselves and that these actions on the hematopoietic progenitor cells are probably not mediated through monocytes and/or lymphocytes.

Stimulation of human natural killer cytotoxicity and protection of mice from infection due to herpes simplex virus by recombinant human leukocyte interferon.

Destruction of virus-infected cells in the absence of antibody (natural killer cytotoxicity [NKC]) has been correlated with resistance to infection due to herpes simplex virus (HSV). Two preparations of human leukocyte interferon produced by recombinant DNA bacteria--IFLrA and IFLrD--were demonstrated to increase NKC in a dose-dependent fashion in vitro. IFLrA was more potent than was IFLrD in this assay. When administered intraperitoneally in combination with human leukocytes to one-week-old mice, IFLrA and IFLrD significantly increased survival (to 47.6% [P less than 0.01] and 23.8% [P less than 0.05], respectively) after an otherwise uniformly lethal challenge of HSV. Differences between interferon immunostimulating activities at similar antiviral concentrations may be important in vivo in animals and in clinical settings.

The anticellular and protein-inducing activities of human gamma interferon preparations are mediated by the interferon.

The anticellular and protein-inducing activities of human gamma interferon preparations are mediated by the interferon.