Recombinant Interferon-alpha Research Articles

Safety of aerosol inhalation of recombinant human interferon alpha 2b injection(P.putida)and excipient hydroxyethyl starch 40 to healthy SD rats

Objective To investigate the effects of recombinant human interferon alpha 2b injection(P.putida)and hydroxyethyl starch inhalation on physiological indexes and tissues and organs of SD rats, such as lungs, trachea and bronchus, and thus to explore the safety of the drug and excipients for inhalation. Methods Rats were randomly divided into two groups(hydroxyethyl starch 40 group and recombinant human interferon alpha 2b group).5 million IU/ml recombinant human interferon alpha 2b injection or 6% hydroxyethyl starch 40 injection were given respectively, which were diluted with 0.9% sodium chloride injection in a radio of 1: 2.Both groups were given aerosol inhalation for 30 minutes once a day for 14 days.Within 14 days after administration, the behavior of rats and the degree of toxicity were continuously observed.During the drug administration and recovery period, the clinical signs of the rats in each group were observed daily, and the body weight and food consumption were measured once a week.At the end of drug administration and the end of recovery period, hematology test, blood biochemical test, gross anatomical observation, organ weight determination, and histopathological examination were performed. Results The mass median aerodynamic diameter(MMAD)of the drug mist was 3.56 μm, and the geometric standard deviation(GSD)was 1.84 μm, which met the particle size requirements of arrival and deposition in the lower respiratory tract via atomization inhalation.No abnormality in clinical signs, body weight and food consumption, hematology and blood biochemical indexes were found in hydroxyethyl starch 40 group and recombinant human interferon alpha 2b group.No abnormal histopathological changes were observed in oral mucosa, tongue, nasal cavity(paranasal sinus), larynx, trachea, main bronchial tube, lung, heart, liver, spleen, kidney, reproductive system and other organs or tissues examined. Conclusion Results suggest the drug particle size of recombinant human interferon alpha 2b injection(pseudomonas)can reach the lower respiratory tract and deposit in the lower respiratory tract.Continuous atomization inhalation for 14 days has a good safety, and the excipient hydroxyethyl starch 40 has a high safety. Key words: Recombinant human interferon alpha 2b; Injection; Aerosol inhalation; Hydroxyethyl starch 40; Safety

Systems of intraganglia and intraneural control over latent alpha-herpesvirus in sensory ganglia of peripheral nervous system

The article presents modern data on interferon‑dependent mechanisms of control of latent alpha herpesvirus in the sensory nerve ganglia of the peripheral nervous system. The data on intraganglionar control mechanisms presented by resident natural killers, natural killer T‑lymphocytes and cytotoxic T‑cells that are part of the satellite cells of the nerve node, and the system of intraneuronal defense contained in the centripetal fibers and presented by cascade system TLR‑3, which leads to the production of pre‑immune interferons‑alpha, ‑beta and ‑lambda with a visorathic action, thus counteracting the centripetal transneural spread of the virus from the periphery to the central nervous system. The joint functioning of the interferon‑dependent systems of intraganglionar and intraneuronal control of alpha herpes viruses ensures the maintenance of the latent state of the pathogen, and the disruption of their work is associated with the risk of reactivation of the virus from latency with the development of clinical symptoms. At the same time, lack of intraganglionar cellular immunity is associated with frequent herpetic eruptions on the skin and mucous membranes, while a deficiency of intraneuronal TLR‑3‑dependent immunity leads to the development of herpes virus lesions of the central nervous system — myelitis, encephalitis, and meningitis. It supports the well‑known clinical fact, that herpetic encephalitis often develops in patients without a history of frequent specific exanthema, and individuals with frequent herpetic eruptions on the skin and mucous membranes may never suffer from viral lesions of the brain and spinal cord. New knowledge about the mechanisms of retaining alpha herpesviruses in the latent state of the sensory ganglia can help in the selection of rational therapy for immunocompromised patients with episodes of herpesvirus neuroinfections with the inclusion of drugs of natural, recombinant and lymphoblastoid human alpha interferons and various inducers of interferonogenesis of chemical or natural origin.The article will be useful, first of all, to neurologists, neurosurgeons, infectious diseases specialists, clinical immunologists, pathologists, physiologists.

High level extracellular production of recombinant human interferon alpha 2b in glycoengineered Pichia pastoris: culture medium optimization, high cell density cultivation and biological characterization.

The present study was aimed at design of experiments (DoE)- and artificial intelligence-based culture medium optimization for high level extracellular production of a novel recombinant human interferon alpha 2b (huIFNα2b) in glycoengineered Pichia pastoris and its characterization. The artificial neural network-genetic algorithm model exhibited improved huIFNα2b production and better predictability compared to response surface methodology. The optimized medium exhibited a fivefold increase in huIFNα2b titre compared to the complex medium. A maximum titre of huIFNα2b (436mgl-1 ) was achieved using the optimized medium in the bioreactor. Real-time capacitance data from dielectric spectroscopy were utilized to model the growth kinetics with unstructured models. Biological characterization by antiproliferative assay proved that the purified recombinant huIFNα2b was biologically active, exhibiting growth inhibition on breast cancer cell line. Culture medium optimization resulted in enhanced production of huIFNα2b in glycoengineered P. pastoris at both shake flask and bioreactor level. The purified huIFNα2b was found to be N-glycosylated and biologically active. DoE-based medium optimization strategy significantly improved huIFNα2b production. The antiproliferative activity of huIFNα2b substantiates its potential scope for application in cancer therapy.

Let-7f microRNA expression within established bladder cancer subtypes and upregulation with recombinant interferon-alpha.

462 Background: Let7f is a microRNA (miR) that may act as a tumor suppressor in multiple human malignancies. In patients with BCG-refractory non-muscle invasive bladder cancer, detectable let7f levels in the urine of patients undergoing treatment with adenoviral vector-mediated IFNα gene therapy (clinical trial setting) were found to be prognostic of treatment response at 12 months. Here we sought to define the association of let7f with clinical bladder cancer phenotypes within The Cancer Genome Atlas (TCGA) cohort and characterize the role of let7f in preclinical models. Methods: Using miRNA sequencing data from the published TCGA 2018 bladder cohort (n = 412), let7f miR expression was characterized and stratified by molecular subtype (basal-squamous, luminal, luminal-infiltrated, luminal-papillary, and neuronal). Kaplan-Meier analyses were performed to compare survival (upper vs lower median, by let7f expression level). The murine urothelial carcinoma cell line MB49 was cultured in colonies of 105 and in vitro administrations of 100U/mL of recombinant mu-IFNα were performed with cells harvested at 0, 2, 8, 12, and 24 hours. Let7f miR expression levels were then determined by qRT-PCR. Results: Within the TCGA cohort, let7f expression levels were found to be lower in the more aggressive basal-squamous (median = 12.8), luminal-infiltrated (median = 12.9), and neuronal (median = 12.4) subtypes relative to the luminal (median = 13.2) and luminal-papillary (median = 13.3) subtypes (p < 0.001). Survival analysis demonstrated a trend towards improved survival in the high let7f expressers, but this did not reach statistical significance. In MB49 cells treated with mu-IFNα, there was a time-dependent increase in let7f miR expression suggesting that IFNα may play a role in upregulation of this potential tumor suppressor. Conclusions: Let7f miR is downregulated in more aggressive subtypes of MIBC. IFNα upregulates let7f expression in MB49 cells, and the inability to do so may comprise an escape mechanism for IFNα-treated tumors. Let7f miR may serve as a novel biomarker to identify aggressive phenotypes and treatment failures for bladder tumors treated with IFNα-based gene therapy.

Corrective Effect of Gentabiferon-S on Weaned Piglet Immune Status and Its Effectiveness in Prevention of Intestinal Infections

The objective of the survey was to study the corrective effect of the Gentabiferon-S preparation on the immune status of the weaned piglets and its effectiveness in prevention of intestinal infections. A positive effect of the preparation on the morphological and immunological variables expressing the increased levels was ascertained. The red blood cell count, the relative lymphocyte count, the relative and absolute T–lymphocyte counts, the relative and absolute B-cell counts, the complement system activity, the lysozyme level, the neutrophil phagocytic activity, the number of phagocytes, the phagocytic index, the spontaneous and stimulated NBT test, the neutrophil activation reserve and index, and the amount of total immunoglobulins increased by 10.8, 16.5, 16.8 and 14.0%, 21.0 and 20.0%, 23.5, 66.8, 15.9, 38.2, 16.2, 11.0 and 21.7%, 13.3 and 17.1%, and 9.7%, respectively. A high (100%) effectiveness of the preparation in prevention of the intestinal infections is ensured by the presence of the antibiotic with broad-spectrum antimicrobial activity and the porcine recombinant alpha and gamma interferon.

Long-Term Efficacy and Safety of Recombinant Interferon Alpha-2 Vs. Hydroxyurea in Polycythemia Vera: Preliminary Results from the Three-Year Analysis of the Daliah Trial - a Randomized Controlled Phase III Clinical Trial

BackgroundRecombinant Interferon Alpha-2a (r-IFNα) is a potent immunomodulating agent, which has been used off-label for the treatment of polycythemia vera (PV) for more than three decades and has been demonstrated to induce high rates of clinical, hematological and molecular responses. Only few studies have compared efficacy and safety of r-IFNα vs. hydroxyurea (HU), which is considered first line therapy for PV patients > 60 years in most parts of the world. However, recent studies have provided encouraging results for the treatment of PV with r-IFNα compared to HU irrespective of age (R. Hoffmann 2016; H. Gisslinger 2018).AimsTo examine the difference in efficacy and safety of low-dose r-IFNα in PV patients ≤ 60 or > 60 years of age compared to HU > 60 years of age.MethodsNinety newly diagnosed or previously phlebotomized PV patients only (WHO 2008) were enrolled in the DALIAH trial (NCT01387763). All patients provided written informed consent. Patients ≤ 60 years were randomized (I:I) to r-IFNα-2a (Pegasys®) or to r-IFNα-2b (PegIntron®) whereas patients > 60 years were randomized (I:I:I) to either r-IFNα-2a, r-IFNα-2b or to HU. The starting dose of r-IFNα-2a and r-IFNα-2b was 45 or 35 µg/week, respectively. The HU dose was 500 to 2000 mg/day. Patients randomized to r-IFNα who presented with major thrombosis or platelets > 1500 109/L received HU from inclusion and until normalization of the platelet count. Efficacy assessment consisted of the clinicohematological and the molecular response rates by intention to treat analysis (ITT) using the European Leukemia Net (ELN) 2009 criteria. JAK2V617F analysis was performed by qPCR. Groups were compared by Fisher's Exact Test.ResultsThree-year analysis was available in all but five patients (n=85) at time of abstract submission (Table 1). The analysis was performed after a median of 36 months (range: 33-39 months). The median treatment dose was 684 mg/day (IQR: 131 - 942) for HU, 51 μg/week (IQR: 30-90 μg/week) and 54 μg/week (IQR: 30-66 μg/week) for r-IFNα-2a age ≤ 60 and > 60, respectively, and 41 μg/week (IQR: 29-45 μg/week) and 36 μg/week (IQR: 31-37 μg/week) for r-IFNα-2b age ≤ 60 and > 60.The overall clinicohematological response rate (ORR) was 68% (13/19) for HU, 42% (14/33) for r-IFNα ≤ 60 years and 39% (13/33) for r-IFNα > 60 years. The partial clinicohematological response rate (PHR) and the complete clinicohematological response rate (CHR) was 53% (10/19) and 16% (3/19) for HU, 9% (3/33) and 33% (11/33) for r-IFNα ≤ 60 years and 9% (3/33) and 30% (10/33) for r-IFNα > 60 years. Neither the ORR, CHR nor the PHR was significantly different between the three groups. Maintenance of CHR from first occurrence to data analysis after 36 months was 11% (2/19) for HU, 21% (7/33) for r-IFNα ≤ 60 years and 18% (6/33) for r-IFNα > 60 years.Forty-seven JAK2V617F positive patients were available for molecular response analysis after 36 months of therapy. A partial molecular response (PMR) was detected in 21% (4/19) of HU treated patients and in 24% (7/29) of r-IFNα treated patients ≤ 60 years and 18% (6/33) of r-IFNα > 60 years. Notably, 7% (2/29) of the r-IFNα treated patients ≤ 60 years obtained a complete molecular response (CMR). The median JAK2V617F reduction from baseline was 38% (IQR: 31-63%) for HU, 79% (IQR: 59-92%) for r-IFNα ≤ 60 years and 73% (IQR: 49-97%) for r-IFNα > 60 years. There was no statistically significant difference in the PMR between groups.Discontinuation of treatment for any reason after 36 months of therapy was 21% (4/19) for HU, 52% (17/33) for r-IFNα ≤ 60 years and 45% (15/33) for r-IFNα > 60 years. Toxicity related discontinuation was 5% (1/19) for HU and 30% (10/33) for both r-IFNα ≤ 60 and > 60 years. Grade 3-4 AEs occurred in 32% (6/19) of HU treated patients, 27% (9/33) in r-IFNα treated patients ≤ 60 years and in 42% (14/33) r-IFNα treated patients > 60 years. SAEs were reported in 21% (4/19) for HU, 9% (3/33) for r-IFNα ≤ 60 years and 24% (8/33) for r-IFNα > 60 years. The numbers of grade 3-4 AEs as well as SAEs were comparable between groups.ConclusionAfter 36 months of therapy CHR was non-significantly higher in PV patients treated with r-IFNα compared to HU by ITT, irrespective of age. Also, maintenance of CHR was longer for r-IFNα. However, ORR was non-significantly higher for HU. PMR was almost similar between the three groups but the median JAK2V617F reduction was greater for r-IFNα. Toxicity related discontinuation from study therapy was higher for r-IFNα compared to HU. DisclosuresStentoft:Bristol-Myers Squibb: Research Funding; Merck Sharp&Dohme: Research Funding. Hasselbalch:Novartis: Research Funding.

Attach great importance to innovative research and clinical application of recombinant human interferon alpha 1b in pediatrics

As the first class Ⅰ novel genetic engineering medical product in China, recombinant human interferon alpha 1b (IFNα1b) shows unique biological significance and clinical value.Especially in the field of pediatrics, IFNα1b has been used in the treatment of viral pneumonia, viral hepatitis, bronchiolitis, herpetic angina, hand-foot-mouth disease and some tumors in children and shown definite efficacy and good safety.It is also used in the prevention and treatment of virus-related wheezing and major infectious diseases, such as influenza.With a greater application prospect, IFNα1b plays an important role in promoting children′s health in China.Now, IFNα1b′s gene and molecular characteristics, new delivery methods, and the frontier clinical research will be discussed. Key words: Interferon α1b; Child; Virus; Inhalation; Wheezing; Influenza

Cepeginterferon alfa-2b in the treatment of chronic myeloproliferative diseases

A comparative evaluation of the effectiveness of different therapeutic strategies in patients with polycythemia vera (PV) and essential thrombocythemia (ET). Patients with PV or ET, diagnosed according to the criteria WHO 2016 were included in the study. The primary endpoint - 6 months of therapy (clinical-hematological and molecular responses). The secondary endpoint - 12 months of therapy (clinico-hematologic, molecular, histological responses). Sixty three patients were included in the analysis: the first group consisted of 33 patients who received the therapy with ce-pegiterferone alpha-2b (ce-pegalpha-INF-α-2b), 10 of them received previous treatment; the second group - 23 patients btained hydroxycarbamide; the third group - 7 patients were treated with recombinant interferon alpha therapy (rINFα). In comparison groups, differences in age were revealed: patients receiving hydroxycarbamide therapy were older. Phlebotomy occurred in 36% of patients in the first group, 9% in the second group, and 14% in the third group. By the 6th month of therapy, 43% of the patients receiving the ce-pegalpha-INF-α-2b had complete clinical-hematologic response, 36% had partial clinical-hematologic remission and stabilization of the disease was established in 21% cases. No disease progression occured. By the 12th month of therapy, statistically significant differences in terms of efficacy between the different therapeutic groups (p = 0.2462, Fisher's exact test). In all three groups, the allelic load of JAK2V617F decreased: from 50 to 19%, from 22.3 to 15.8%, from 50 to 7.19%, respectively. The lower the allele load positively correlated with better response to therapy, which was observed in all analyzed groups. Hematologic adverse events (AEs) were more frequently observed in patients receiving ce-pegalpha-INF-α-2b therapy. Local reactions developed on 3-7 days of therapy as a hyperemic macula at the injection site. Both these reactions and hair loss did not influence on patient's condition. In the second group (patients with hydroxycarbamide therapy) there were changes in the skin and mucous membranes: dry skin, stomatitis, and in older patients new keratomas appeared. The flu-like syndrome was the most common adverse event associated with the therapy of ce-pegalpha-INF-α-2b, which fully relived during the first month of therapy. There was only one case with the flu-like syndrome we observed at the 11th month of therapy. As a rule, the biochemical blood test changes did not influence on patient's condition, were mostly associated with dietary violations, had a tendency to self-resolution and did not require medical interventions. Serious AEs were reported in one case - pulmonary embolism in a patient treated with rINFα. The reasons for the therapy discontinue in group 1: toxic hepatitis, intolerance, by the request of the patient, inadequate efficacy of therapy; in group 2: skin toxicity, in group 3: thromboses. Treatment of ce-pegalpha-INF-α-2b in patients with PV and ET is highly effective - the most patients pbtained clinical and hematological responses. There were no statistically significant differences in these parameters in comparison with hydroxycarbamide and rINFα. The use of the ce-pegalpha-INF-α-2b had an acceptable safety profile. The estimated therapeutic dose should be calculated according to body weight. To reduce the frequency of hematologic AE, titration of the drug dose is required.

PO-040 Development of a tunable form of interferon alpha for in vivo cancer gene therapy

IntroductionThe immune system is a double-edge sword in cancer. On the one hand, it exerts immunosurveillance to eradicate transformed cells that occasionally appear in the body; on the other hand, cancer cells can recruit immune cells endowed with pro-tumorigenic activity.Our lab previously developed a strategy for targeted gene-based delivery of interferon alpha (IFNa) to tumours by tumour infiltrating monocytes/macrophages, which induces robust anti-cancer responses in several experimental models without inducing strong IFN responses in normal tissues as compared to systemic administration of recombinant IFNa. Whereas a sustained output could ensure long-term protection from tumour recurrence, it may raise concerns for long-term side effects, especially in case of cancer eradication.To overcome this issue, we are developing inducible strategies to control the amount of IFNa secreted in the tumour microenvironment.Material and methodsBy fusing a destabilising domain (DD) to a protein of interest (POI) the former can confer its instability to the latter. This destabilisation can be rescued in a reversible and dose dependent manner with the addition of a small molecule specifically binding to the DD. To apply this technology to our strategy we have designed and in vitro tested different fusion proteins of IFNa (DD-IFNa). We also developed improved DD-IFNa with the addition of flexible and/or cleavable linkers and selected them for their capacity to be stabilised in a dose dependent manner in presence of their specific ligand in vitro.Results and discussionsThrough this approach, we have identified effective fusion proteins with low basal activity and high fold induction upon ligand treatment. These novel tunable forms of IFNa are functional and their specific activity are comparable to the wild type cytokine in inducing IFN responsive genes.Based on these promising in vitro results we are now translating these new platforms in vivo to test their efficacy in inducing anti-tumour responses in melanoma, colon and glioma models of cancer.ConclusionIn the perspective of clinical translation our approach can be used in the future to switch on/off the levels of IFNa in a tunable and personalised fashion for cancer eradication.

NEW DISCOVERIES IN THE MECHANISMS OF INTERFERON-DEPENDENT CONTROL OF LATENT ALPHA-HERPESVIRUS IN SENSORY GANGLIA. REVIEW

Relevance. The study of the mechanisms of immune surveillance over latent alpha herpesvirus in the sensory nerve ganglia is an important task of clinical immunology, since the data obtained during such studies may help to develop effective approaches to immunotherapy of recurrent and/or severe forms of herpes infections by restoring lost immune control over latent pathogens. Objective: to conduct a systematic review of the results of studies devoted to the study of mechanisms of intraganglionar and intraneuronal protection over latent alpha herpesvirus in sensory nerve ganglia, to develop a modern concept of immune surveillance in human alpha herpesvirus infections. Methods. Review of scientific publications in the international electronic science databases PubMed, Embase and Scopus was conducted on the keywords «alpha herpesviruses», «Toll-like receptor type 3», «natural killers», «natural killer T-cells», «intraganglionar immunity», «intraneuronal immunity» for the full time available (1972-2014). Results. The article presents modern data on interferon-dependent mechanisms of control of latent alpha herpesvirus in sensory nerve ganglia. The system of the intraganglionar control presented by resident natural killers, natural killer T-lymphocytes and cytotoxic CD8+ T-cells that are part of the satellite cells of the nerve node, and the system of intraneuronal defense contained in the centripetal fibers and presented by cascade system TLR-3 , the activation of which promote the production of interferons type I. Genetic or acquired abnormalities in intraganglionar and intraneuronal immunity systems are the cause of the development of recurrent and/or severe forms of infections caused by alpha herpesviruses in humans. Preparations of natural and recombinant alpha interferons and interferonogens are potentially useful agents in the target immunotherapy of disorders in intraganglionar and intraneuronal immunity in immunocompromised patients with severe forms of alpha herpesvirus infections. Conclusions. In the human body there are highly organized intraganglionar and intraneirononal immune control systems for latent alpha herpesviruses. The activity of these systems ensures the long stay of the virus in a latent state in the sensory nerve ganglia, and the disturbanses of the implementation of such mechanisms is responsible for reactivation of the virus from the latent state with subsequent damage to the skin, mucous membranes and/or the central nervous system.

NEW DISCOVERIES IN THE MECHANISMS OF INTERFERON-DEPENDENT CONTROL OF LATENT ALPHA-HERPESVIRUS IN SENSORY GANGLIA. REVIEW

Relevance. The study of the mechanisms of immune surveillance over latent alpha herpesvirus in the sensory nerve ganglia is an important task of clinical immunology, since the data obtained during such studies may help to develop effective approaches to immunotherapy of recurrent and/or severe forms of herpes infections by restoring lost immune control over latent pathogens.
 Objective: to conduct a systematic review of the results of studies devoted to the study of mechanisms of intraganglionar and intraneuronal protection over latent alpha herpesvirus in sensory nerve ganglia, to develop a modern concept of immune surveillance in human alpha herpesvirus infections.
 Methods. Review of scientific publications in the international electronic science databases PubMed, Embase and Scopus was conducted on the keywords «alpha herpesviruses», «Toll-like receptor type 3», «natural killers», «natural killer T-cells», «intraganglionar immunity», «intraneuronal immunity» for the full time available (1972-2014).
 Results. The article presents modern data on interferon-dependent mechanisms of control of latent alpha herpesvirus in sensory nerve ganglia. The system of the intraganglionar control presented by resident natural killers, natural killer T-lymphocytes and cytotoxic CD8+ T-cells that are part of the satellite cells of the nerve node, and the system of intraneuronal defense contained in the centripetal fibers and presented by cascade system TLR-3 , the activation of which promote the production of interferons type I. Genetic or acquired abnormalities in intraganglionar and intraneuronal immunity systems are the cause of the development of recurrent and/or severe forms of infections caused by alpha herpesviruses in humans. Preparations of natural and recombinant alpha interferons and interferonogens are potentially useful agents in the target immunotherapy of disorders in intraganglionar and intraneuronal immunity in immunocompromised patients with severe forms of alpha herpesvirus infections.
 Conclusions. In the human body there are highly organized intraganglionar and intraneirononal immune control systems for latent alpha herpesviruses. The activity of these systems ensures the long stay of the virus in a latent state in the sensory nerve ganglia, and the disturbanses of the implementation of such mechanisms is responsible for reactivation of the virus from the latent state with subsequent damage to the skin, mucous membranes and/or the central nervous system.

Philadelphia chromosome-negative classical myeloproliferative neoplasms: revised management recommendations from European LeukemiaNet.

This document updates the recommendations on the management of Philadelphia chromosome-negative myeloproliferative neoplasms (Ph-neg MPNs) published in 2011 by the European LeukemiaNet (ELN) consortium. Recommendations were produced by multiple-step formalized procedures of group discussion. A critical appraisal of evidence by using Grades of Recommendation, Assessment, Development and Evaluation (GRADE) methodology was performed in the areas where at least one randomized clinical trial was published. Seven randomized controlled trials provided the evidence base; earlier phase trials also informed recommendation development. Key differences from the 2011 diagnostic recommendations included: lower threshold values for hemoglobin and hematocrit and bone marrow examination for diagnosis of polycythemia vera (PV), according to the revised WHO criteria; the search for complementary clonal markers, such as ASXL1, EZH2, IDH1/IDH2, and SRSF2 for the diagnosis of myelofibrosis (MF) in patients who test negative for JAK2V617, CALR or MPL driver mutations. Regarding key differences of therapy recommendations, both recombinant interferon alpha and the JAK1/JAK2 inhibitor ruxolitinib are recommended as second-line therapies for PV patients who are intolerant or have inadequate response to hydroxyurea. Ruxolitinib is recommended as first-line approach for MF-associated splenomegaly in patients with intermediate-2 or high-risk disease; in case of intermediate-1 disease, ruxolitinib is recommended in highly symptomatic splenomegaly. Allogeneic stem cell transplantation is recommended for transplant-eligible MF patients with high or intermediate-2 risk score. Allogeneic stem cell transplantation is also recommended for transplant-eligible MF patients with intermediate-1 risk score who present with either refractory, transfusion-dependent anemia, blasts in peripheral blood > 2%, adverse cytogenetics, or high-risk mutations. In these situations, the transplant procedure should be performed in a controlled setting.

Efficacy of nystatin vaginal effervescent tablets and nifuratel tablet combined with recombinant human interferon alpha 2b suppository in the treatment of candida vaginitis

Objective To explore the efficacy of nystatin vaginal effervescent tablets, nifuratel tablet combined with recombinant human interferon alpha 2b suppository in the treatment of candida vaginitis. Methods 124 patients with candida vaginitis were selected as the research subjects.The patients were randomly divided into two groups according to different treatment.The control group was treated with nystatin vaginal effervescent tablets + nifuratel tablet, the study group was given recombinant human interferon alpha 2b suppository on the basis of the control group.The symptoms, clinical curative effect, quality of life score, adverse reaction rate were observed and compared in the two groups. Results There were no statistically significant differences between the two groups in symptom and physical signs before treatment(all P>0.05). After treatment, the vulvar itching, burning pain, abnormal leucorrhea, redness symptoms and signs of the study group were (0.95±0.44)points, (1.02±0.51)points, (1.13±0.62)points, (1.09±0.56)points, which were significantly lower than those of the control group[(1.42±0.98)points, (1.28±1.02)points, (1.38±1.17)points, (1.37±1.12)points, t=2.529, 2.512, 2.505, 2.618, all P<0.05]. The total effective rate of the study group was 95.2%, which was significantly higher than 74.2% of the control group (Z=-2.839, P<0.05). After treatment, the quality of life scores of physiological function, bodily pain, mental health, emotional function and social function in the study group were (80.16±9.23)points, (84.22±9.95)points, (87.49±10.46)points, (89.27±12.35)points, (88.21±11.74)points, which were significantly higher than those in the control group[(67.58±5.34)points, (71.49±7.28)points, (73.52±7.61)points, (76.83±8.32)points, (75.51±8.16) points, t=9.728, 8.577, 9.036, 7.039, 7.537, all P<0.05]. The incidence rate of adverse reactions of the study group was significantly lower than that of the control group(χ2=10.978, P<0.05). Conclusion Nystatin vaginal effervescent tablets, nifuratel tablet combined with recombinant human interferon alpha 2b suppository in the treatment of candida vaginitis can significantly improve clinical symptoms, improve the therapeutic effect, and improve the quality of life of patients with significant clinical value, it is worthy of reference. Key words: Candidiasis, vulvovaginal; Nystatin; Nifuratel; Interferon alpha 2b

Recombinant interferon alpha 2b in rheumatoid arthritis: good antigen for rheumatoid arthritis antibodies.

Aim of the studyInterferon alpha-induced arthritis and activation of the type 1 interferon pathway during rheumatoid arthritis (RA) has been well documented but the underlying mechanism remains unclear. This study addressed the binding specificity of antibodies with recombinant interferon alpha 2b (rIFN α-2b) in sera from different RA patients. Utilization of anti-hrIFN α-2b antibodies as a probe for estimation of interferon α-2b concentration in RA patients’ synovial fluid (SF) was also investigated.Material and methodsBinding specificities of antibodies from the sera of 60 RA patients and 35 controls subjects were studied by direct binding, inhibition ELISA, and quantitative precipitation titration. Inhibition ELISA was also used to estimate patients’ SF interferon α-2b concentrations.ResultsRA IgG from patients’ sera showed strong recognition to hrIFN α-2b in comparison to commercially available interferon (IFN α-2b) (p < 0.05) or the gene encoding this interferon (IFN α-2b gene) (p < 0.05). The affinity of RA antibodies for rIFN α-2b (1.10 × 10–7 M) was found to be high as assessed by Langmuir plot. No significant difference in the level of interferon α in the SF of RA patients was observed as compared to the healthy controls.ConclusionsrIFN α-2b presents unique epitopes that might explain the possible antigenic role in the induction of RA antibodies and anti-rIFN α-2b antibodies represent an alternative immunological probe for the estimation of interferon α in the SF of RA patients.

De-immunized and Functional Therapeutic (DeFT) versions of a long lasting recombinant alpha interferon for antiviral therapy

Interferon α (IFN-α) exerts potent antiviral, immunomodulatory, and antiproliferative activity and have proven clinical utility in chronic hepatitis B and C virus infections. However, repeated IFN-α administration induces neutralizing antibodies (NAb) against the therapeutic in a significant number of patients. Associations between IFN-α immunogenicity and loss of efficacy have been described.So as to improve the in vivo biological efficacy of IFN-α, a long lasting hyperglycosylated protein (4N-IFN) derived from IFN-α2b wild type (WT-IFN) was developed. However, in silico analysis performed using established in silico methods revealed that 4N-IFN had more T cell epitopes than WT-IFN. In order to develop a safer and more efficient IFN therapy, we applied the DeFT (De-immunization of Functional Therapeutics) approach to producing functional, de-immunized versions of 4N-IFN.Using the OptiMatrix in silico tool in ISPRI, the 4N-IFN sequence was modified to reduce HLA binding potential of specific T cell epitopes. Following verification of predictions by HLA binding assays, eight modifications were selected and integrated in three variants: 4N-IFN(VAR1), (VAR2) and (VAR3). Two of the three variants (VAR1 and VAR3) retained anti-viral function and demonstrated reduced T-cell immunogenicity in terms of T-cell proliferation and Th1 and Th2 cytokine levels, when compared to controls (commercial NG-IFN (non-glycosylated), PEG-IFN, WT-IFN and 4N-IFN).It was previously demonstrated that N-glycosylation improved IFN-α pharmacokinetic properties. Here, we further reduce immunogenicity as measured in vitro using T cell assays and cytokine profiling by modifying the T cell epitope content of a protein (de-immunizing). Taking into consideration the present results and previously reported immunogenicity data for commercial IFN-α2b variants, 4N-IFN(VAR1) and 4N-IFN-4N(VAR3) appear to be promising candidates for improved IFN-α therapy of HCV and HBV.

The Effect of Initial Molecular Profile on Response to Recombinant Interferon Alpha (rIFNα) Treatment in Early Myelofibrosis

IntroductionSuccessful treatment of early myelofibrosis (eMF), defined as patients (pts) with low or intermediate-1 (int-1) risk and ≥15% hematopoietic foci in the marrow biopsy, with rIFNα has been reported, but effects of driver and epigenetic mutations on outcomes were not considered. We now report results of our single center phase II study of 30 pts with primary (PM) or secondary (SMF) eMF, correlating the baseline molecular profile with response to rIFNα treatment.MethodsWe used WHO 2008 and IWG-MRT criteria for the diagnosis of PM and post-polycythemia vera MF (pPV MF) or post-essential thrombocythemia MF (pET MF), respectively. Prognosis was assessed using the Dynamic International Prognostic Scoring System (DIPSS). Response classifications were complete response (CR), partial response (PR), clinical improvement (CI), stable disease (SD), progressive disease (PD). Evaluation included history, examination, spleen size (cm below left costal margin), CBC and differential counts, routine chemistries, liver, renal, and thyroid function tests, marrow biopsy, cytogenetic and molecular genetic evaluations. Marrow specimens were stained routinely for morphology, reticulin, and collagen; fibrosis was graded using the European consensus system.JAK2, CALR and MPL mutations were detected and quantified by amplicon based next generation sequencing (NGS). NGS of 45 genes recurrently mutated in myeloid malignancies was performed on 25 of 30 baseline specimens, including ASXL1, IDH1/2, EZH2 and SRSF2, previously defined as high molecular risk (HMR) genes.Descriptive statistics were calculated and Fisher’s exact test used to explore the relationship between mutations and various results. Pts received either rIFNα-2b or peg-rIFNα-2a starting at a 5x105 to 1x106 units SC 3X wk or 45 to 90 mcg weekly, respectively; dose increase dependent upon tolerance and/or response.ResultsOf 96 pts with PM or SMF, 30 met study criteria, including 16 women and 14 men (med. dx age 58 yrs). Twenty-one had PM, 7 pPV MF, and 2 pET MF. Twenty-one pts were low risk, 9 int-1. Twenty-two were JAK2V617F(+), 6 CALR(+), and 2 MPL(+). All 7 pPV MF pts were JAK2(+). One pET MF was JAK2(+) and 1 CALR(+). Of PM pts, 14 were JAK2(+), 5 CALR(+), and 2 MPL(+).Med. rx duration has been 5.6 yrs and is ongoing. Clinical toxicity, grade 1 or 2, included mild depression, dry skin, cough, and myalgia. Hem. toxicity included anemia (grade 1-2: 17 pts), leukopenia (grade 1-2: 9 pts), and thrombocytopenia (grade 1-2: 10; grade 3: 2 pts). In these cases, rIFNα dose was temporarily reduced/interrupted until cytopenias resolved. One pt developed hyperthyroidism; treatment was discontinued.Of 30 pts, 2 (7%) had CR, 9 (30%) PR, 4 (13%) CI, 7 (23%) SD. Four (13%) had PD, and 4 (13%) died. Nine of 21 low risk pts had CR or PR, compared with 2 of 9 int-1. A small spleen at baseline was associated with better response in the absence of HMR abnormalities. Of 18 pts with slight or no splenomegaly, 11 had CR or PR, and of the non-responders, 2 had a HMR. Of 12 pts with significant or moderate splenomegaly, none had CR or PR, including 2 with a HMR. Follow up biopsies in 25 pts (83%) a reduction in reticulin fibrosis in 5, increase in 9, and stable in 10. Decrease in cellularity occurred in 12. There was no correlation between decrease in JAK2 allele burden, response, fibrosis, or spleen size.There was no correlation between driver mutations and response (Table). Pts with a HMR had worse responses; of 4 such pts, 0 responded, 1 remained stable, 1 progressed and 2 died. Specifically, pts with ASXL1 mutations seemed to do worse than those without it, although sample size precludes a definitive statement (p=0.19).ConclusionThis is the first study considering the effect of baseline mutation status on response to rIFNα treatment in eMF pts. 73% of pts improved or remained stable regardless of initial low or int-1 score or driver mutation, but spleen size and HMR adversely affected treatment response.Although baseline driver mutation status was not prognostically useful, a molecular profile should be obtained initially to identify HMR mutations, which negatively affect response to treatment including rIFNα. In 73% of selected cases, low dose rIFNα in eMF resulted in marrow reversion, regression of splenomegaly or disease stability with tolerable toxicity. Early treatment of MF with rIFNα in pts without HMR may prevent the development of marked splenomegaly, anemia, and florid myelofibrosis. [Display omitted] DisclosuresSilver:Incyte: Membership on an entity’s Board of Directors or advisory committees. Ritchie:Pfizer: Honoraria; Incyte: Speakers Bureau; Celgene: Speakers Bureau; Novartis: Honoraria, Research Funding; Arian: Speakers Bureau. Roboz:Agois: Consultancy; Amgen: Consultancy; Amphivena: Consultancy; Astex: Consultancy; AstraZeneca: Consultancy; Boehringer Ingelheim: Consultancy; Celator: Consultancy; Celgene: Consultancy; CTI Biopharma: Consultancy; Genoptix: Consultancy; Janssen Pharamaceuticals: Consultancy; Junno: Consultancy; MedImmune: Consultancy; MEI Pharma: Consultancy; Novartis: Consultancy; Onconova: Consultancy; Pfizer: Consultancy; • Roche Pharmaceuticals: Consultancy; Sunesis: Consultancy; Teva: Consultancy. Orazi:Novartis: Honoraria. Tam:Millennium Pharmaceuticals, Inc.: Consultancy. Cross:Novartis: Consultancy, Honoraria, Research Funding, Speakers Bureau.

Experience using recombinant interferon-alpha with taurine therapy of congenital cytomegalovirus infection in children

Cytomegalovirus (CMV) belongs to the most frequent causes of intrauterine infection and can cause birth defects and severe manifestations of the disease fatal. In most cases the infection is latent, but there is a high risk of developing late manifestations. For the prevention of vertical transmission of CMV infection are needed early prenatal diagnosis of infection in a pregnant woman with the definition of the extent of its activity and conduct of antiviral therapy. When intravenous anti-CMV immunoglobulin observed rapid clinical efficacy. However, the titer of circulating antibodies in the child’s body is rapidly reduced, determines the duration of the protective effect of the drug to 4-5 weeks. Since the suppository form of interferon-α with taurine has been used successfully in obstetric practice, we have examined its effectiveness in the treatment of 11 outpatient children aged 1 month to 3 years with congenital CMV infection, 3 of them received specific immunoglobulin baby immediately after birth. The drug was administered rectally by prolonged scheme for 3 months 2 times a day every 12 hours within 10 days, followed by 1 suppository 2 times a day every other day (3 times a week). The therapy in all children experienced rapid resolution of symptoms CMV: normalization of the size of lymph nodes, liver and spleen, blood count, serum ALT, urinalysis, as well as the disappearance of CMV DNA in blood and IgM antibodies to CMV infection after treatment. During 12 months of follow-up episodes of reactivation of CMV infection in any of the observed child is not mentioned.

Removing a Cystein Group On Interferon Alpha 2b at Position 2 and 99 does Not Diminish Antitumor Activity of the Protein, Even Better

Interferon alpha 2b is the only standard therapeutic protein for hepatitis virus infections. Further study demonstrated that this protein also posseses antitumor activity in several cancerous organs. One main pathway of this antitumor activity is mediated through antiproliferation as well as proapoptotic effects. Previously, we have successfully developed recombinant human interferon alpha 2b (rhIFNα2b) by using a synthetic gene. In addition, two mutein forms of rhIFNα2b were generated to improve the characteristics of this protein. Two point mutations showed better pharmacokinetic profiles than one point mutation as well as the native form. In the present study, this mutein form was studied for ist antitumor effect in vitro using HepG2 cells. As a comparison, the native form as well as a commercial rIFNα2b were used. Several parameters were investigated including the MTT assay, cell viability test, cell cycle using flow cytometric analysis, and the genes and protein expressions involved in cell growth. The latest was observed to study the mechanism of rhIFNα2b. There was no significant difference in the MTT assay and cell viability after cells were treated with both forms of rhIFNα2b. However, the mutein rhIFNα2b tended to show better proapoptotic activity reflected by flow cytometric data, protein expression of pSTAT1, and DNA expression of caspase 3.

Recombinant interferon alpha 2b for ocular surface squamous neoplasia: An efficient and cost-effective treatment modality in Asian Indian patients.

Purpose:The purpose was to study the efficacy of interferon alpha 2b (INF α2b) in the treatment of ocular surface squamous neoplasia (OSSN) and analyze its cost-effectiveness in India.Study Design:This was a retrospective study of thirty patients with OSSN treated with topical INF α2b (1 MIU/cc) ± perilesional INF α2b (5 MIU/cc).Results:The tumor involved cornea (n = 9, 30%), conjunctivo-limbal-corneal surface (n = 19, 63%), or bulbar conjunctiva (n = 2, 7%). The mean basal dimension of the tumor was 16 mm. The tumors belonged to Tis (n = 6, 20%) or T3 (n = 24, 80%) based on the American Joint Committee Classification, 7th edition. In the six patients with Tis, three cycles of topical INF α2b were used for immunoprevention. In the remaining 24 patients, INF α2b was advised for immunoreduction, but served as immunotherapy with 100% tumor regression in 22 (92%) cases, and resulted in 95% immunoreduction in 2 (6%) cases. Complete tumor regression by immunotherapy (n = 22) was achieved with a mean number of three topical INF α2b cycles and two perilesional injections. All these 22 patients received three additional topical INF α2b cycles after complete tumor regression. For immunoreduction (n = 2), both patients received six cycles of topical INF α2b which was three perilesional INF α2b injections. The mean total treatment cost per patient with INF α2b was INR 9164 ($US 137). Based on maximum basal diameter of tumor at presentation, the mean total treatment cost per patient with INF α2b was INR 4866 ($US 73) for eyes with microscopic evidence of tumor residue (n = 6), INR 9607 ($US 143) for tumors ≤10 mm (n = 13), and INR 10,985 ($US 164) for tumors >10 mm (n = 11), with two patients needing additional surgical excision for complete tumor control.Conclusion:INF α2b can be used for immunoreduction, immunotherapy, or immunoprevention of OSSN. INF α2b is a cost-effective treatment modality for OSSN at an average total treatment cost of INR 9164 ($US 137) per patient.

Эффективность рекомбинантного интерферона-альфа в лечении и профилактике ОРВИ

Эффективность рекомбинантного интерферона-альфа в лечении и профилактике ОРВИ