Recombinant Human Interferon-alpha Research Articles

Production of Recombinant Human Interferon alpha 2b (rhIFN?-2b) in genetically engineered strain of Bacillus subtilis

Interferon alpha 2b (IFN?-2b) is a critical therapeutic protein with immunomodulatory properties, widely used in the treatment of various diseases, including viral infections and cancer. This study presents a comprehensive methodology for the cloning and characterization of the IFN?-2b gene, including the identification of point mutations, followed by expression in Bacillus subtilis. The research begins with gene isolation, primer design, RNA extraction, cDNA synthesis, and PCR amplification. Cloning into a T/A vector and subsequent sequencing reveal point mutations within the gene. The transformed construct is expressed in Bacillus subtilis, and the resulting protein is analyzed using SDS-PAGE. The identification of point mutations and successful protein expression in Bacillus subtilis. This study provides valuable insights into the IFN?-2b gene and its potential applications.

Use of Recombinant Human Interferon Alpha in felines infected with FeLV

Abstract. The feline leukemia virus (FeLV) is a retrovirus that belongs to the genus Gammaretrovirus and exclusively affects felids. The virus carries the reverse transcriptase enzyme, which integrates into the host's DNA permanently, leading to different clinical conditions. The main complications caused by FeLV are hematopoietic neoplasms, immune-mediated disorders, neurological alterations and immunosuppression, among other ailments. The disease has no effective treatment, but antiviral and immunomodulatory drugs are still under trial. Using Recombinant Human Interferon Alpha has shown to be helpful in reducing clinical symptoms and controlling opportunistic infections, increasing the quality and life expectancy of cats infected with FeLV. Retroviruses are among the main causes of death in domestic cats. The lack of knowledge about the disease among guardians and growing cat population in Brazil are helping to increase the number of FeLV cases. Although the disease lacks a universal veterinary medical protocol, there are new studies and information available that can help to treat infected animals, improving their quality of life and expectancy and, in some cases, potentially eliminating the virus. This fact clashes with another reality: outdated professionals who are not familiar with the guidelines for Feline Retrovirus Testing and Management from the AAFP (American Association of Feline Practitioners), an international entity that since 1971 has been working to improve feline health by providing evidence-based guidelines for veterinary practices. Population growth and diseases that exclusively affect the species also highlight the need to include a deeper understanding of feline health in the curricula of undergraduate courses, preparing these future professionals to meet the demand for updated veterinarians with sufficient knowledge to care for the species.

Specific sequence mutations in a long-lasting rhIFN-α2b version reduce in vitro and in vivo immunogenicity and increase in vitro protein stability

For decades, recombinant human interferon alpha (rhIFN-α2b) has been used to treat emerging and chronic viral diseases. However, rhIFN-α2b is immunogenic and has a short in vivo half-life. To solve these limitations, two long-lasting hyperglycosylated proteins with reduced immunogenicity were developed and designated as 4N-IFN(VAR1) and 4N-IFN(VAR3).Here, we continue to study the relevant characteristics of these therapeutic candidates. Thus, we demonstrated that both de-immunized IFN versions elicited significantly lower neutralizing antibody responses than the original molecule in HLA-DR1 transgenic mice, confirming our previous in vitro protein immunogenicity data. Also, we found that these biobetters exhibited remarkable stability when exposed to different physical factors that the protein product may encounter during its production process and storage, such as low pH, thermal stress, and repeated freezing/thawing cycles. Taking into consideration our previous and present results, 4N-IFN(VAR1) and 4N-IFN-4N(VAR3) appear to be valuable candidates for the treatment of human viral diseases.

Anti-Viral Activities of Umbilical Cord Mesenchymal Stem Cell-Derived Small Extracellular Vesicles Against Human Respiratory Viruses

The endemic and pandemic caused by respiratory virus infection are a major cause of mortality and morbidity globally. Thus, broadly effective antiviral drugs are needed to treat respiratory viral diseases. Small extracellular vesicles derived from human umbilical cord mesenchymal stem cells (U-exo) have recently gained attention as a cell-free therapeutic strategy due to their potential for safety and efficacy. Anti-viral activities of U-exo to countermeasure respiratory virus-associated diseases are currently unknown. Here, we tested the antiviral activities of U-exo following influenza A/B virus (IFV) and human seasonal coronavirus (HCoV) infections in vitro. Cells were subject to IFV or HCoV infection followed by U-exo treatment. U-exo treatment significantly reduced IFV or HCoV replication and combined treatment with recombinant human interferon-alpha protein (IFN-α) exerted synergistically enhanced antiviral effects against IFV or HCoV. Interestingly, microRNA (miR)-125b, which is one of the most abundantly expressed small RNAs in U-exo, was found to suppress IFV replication possibly via the induction of IFN-stimulated genes (ISGs). Furthermore, U-exo markedly enhanced RNA virus-triggered IFN signaling and ISGs production. Similarly, human nasal epithelial cells cultured at the air-liquid interface (ALI) studies broadly effective anti-viral and anti-inflammatory activities of U-exo against IFV and HCoV, suggesting the potential role of U-exo as a promising intervention for respiratory virus-associated diseases.

Pharmacokinetics Versus In Vitro Antiproliferative Potency to Design a Novel Hyperglycosylated hIFN-α2 Biobetter.

IFN4N is a glycoengineered version of recombinant human interferon alpha 2 (rhIFN-α2) that was modified to exhibit four N-glycosylation sites. It shows reduced in vitro specific biological activity (SBA) mainly due to R23 mutation by N23. However, it has improved pharmacokinetics and led to a high in vivo antitumor activity in mice. In order to prepare a new IFN-based biobetter, this work compares the influence of glycosylation (affecting pharmacokinetics) with the in vitro antiproliferative SBA on the in vivo efficacy. Based on IFN4N, three groups of muteins were designed, produced, and characterized. Group A: variants with the same glycosylation degree (4N) but higher in vitro antiproliferative SBA (R23 restored); group B: muteins with higher glycosylation degree (5N) but similar in vitro antiproliferative activity; and group C: variants with improved glycosylation (5N and 6N) and in vitro antiproliferative bioactivity. Glycoengineering was successful for improving pharmacokinetics, and R23 restoration considerably increased in vitro antiproliferative activity of new muteins compared to IFN4N. Hyperglycosylation was able to improve the in vivo efficacy similarly to or even better than R23 restoration. Additionally, the highest glycosylated mutein exhibited the lowest immunogenicity. Hyperglycosylation constitutes a successful strategy to prepare a novel IFN biobetter.

The Effect of Recombinant Human Interferon Alpha Nasal Drops to Prevent COVID-19 Pneumonia for Medical Staff in an Epidemic Area

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Use of immunostimulants in the successful treatment of a clinical EEHV1A infection in an Asian elephant (Elephas maximus)

Elephant endotheliotropic herpesvirus haemorrhagic disease (EEHV‐HD) poses a significant threat to the captive population of juvenile Asian elephants (Elephas maximus) and also affects elephants in the wild. In human and veterinary medicine, increasing attention is returning to the interferon system, a crucial mediator in the immune response for control of infection. We describe the first reported use of the Zelnate DNA immunostimulant and recombinant human interferon alpha (rhIFNα) as additional medications in the successful treatment of a case of EEHV1A‐HD in a juvenile Asian elephant at a UK zoo. Despite an exponential rise in viraemia to a peak of 2.82×106 viral genomic equivalents/ml, only mild clinical signs developed and the calf survived with no adverse effects attributed to the novel treatments. This case is compared with a previous fatal case within the same herd where Zelnate and rhIFNα were not given.

Clinical characteristics and predictors of the duration of SARS-CoV-2 viral shedding in 140 healthcare workers.

BackgroundEpidemiological and clinical features of patients with COVID‐19 have been reported, but none of them focused on medical staff, and few predictors of the duration of viral shedding have been reported. It is urgent to help healthcare workers prevent and recover quickly from the coronavirus disease 2019 (COVID‐19).MethodsWe enrolled 140 medical workers with COVID‐19 in Wuhan. Epidemiological, demographic, clinical, laboratory, radiological treatment and clinical outcome data were collected, and predictors of the duration of viral shedding were explored through multivariable linear regression analysis.ResultsThe medical staff with COVID‐19 presented mild clinical symptoms and showed a low frequency of abnormal laboratory indicators. All the medical staff were cured and discharged, of whom 96 (68.6%) were female, 39 (27.9%) had underlying diseases, the median age was 36.0 years, and 104 (74.3%) were infected whilst working in hospital. The median duration of viral shedding was 25.0 days (IQR:20.0–30.0). Multivariable linear regression analysis showed reducing viral shedding duration was associated with receiving recombinant human interferon alpha (rIFN‐α) treatment, whilst the prolonged duration of viral shedding correlated with the use of glucocorticoid treatment, the durations from the first symptom to hospital admission and the improvement in chest computed tomography (CT) evidence. Moreover, infected healthcare workers with lymphocytes less than 1.1 × 109/L on admission had prolonged viral shedding.ConclusionMedical staff with timely medical interventions show milder clinical features. Glucocorticoid treatment and lymphocytes less than 1.1 × 109/L are associated with prolonged viral shedding. Early admission and rIFN‐α treatment help shorten the duration of viral shedding.

Clinical and Hematological Follow-Up of Long-Term Oral Therapy with Type-I Interferon in Cats Naturally Infected with Feline Leukemia Virus or Feline Immunodeficiency Virus.

Simple SummaryThe viruses which produce feline leukemia (FeLV) and feline immunodeficiency (FIV) attack cells involved in the immune response. As specific drugs are either nonexistent, produce secondary effects, or are expensive, a therapeutic possibility is using nonspecific immunostimulants, such as human interferon alpha. We used this drug to treat 27 cats infected by FeLV and 31 infected by FIV. All cats were naturally infected and treatment was administered orally by their owners for four months. Participating cats were evaluated in our clinics at mid-treatment (M2), end of treatment (M4), and 4–8 months after the end of treatment (M10). We observed that treatment was well tolerated by the cats (as it did not affect the liver or the kidney functions), and improved most of the parameters analyzed (clinic, anemia, white cell counts, and CD4+/CD8+ ratio) as long as it was administered. However, 4–8 months after it was discontinued, though most animals remained clinically healthy, many of these parameters had rebounded to initial values or values even worse than the initial values. Thus, more studies should be conducted with longer administration of this drug to evaluate tolerability and sustained improvement of diseases produced by these two viruses which may lead to death.Feline leukemia virus (FeLV) and feline immunodeficiency virus (FIV), two of the most important pathogens of cats, produce chronic systemic diseases with progressive death of cells involved in the immune response, ultimately leading to death. Immunostimulants is one of the few alternatives to the symptomatic treatment. In this study, 27 naturally FeLV-infected (FeLV+) and 31 naturally FIV-infected (FIV+) cats were administered orally by their owners 60 IU/day of recombinant human interferon alpha (rHuIFN-α) for four months in alternate weeks. Clinical status was evaluated and blood samples collected at four different visits or months (M): pretreatment (M0), mid-treatment (M2), end of treatment (M4), and 4–8 months after end of treatment (M10). Most cats ostensibly improved their clinical status, and many became asymptomatic. rHuIFN-α treatment improved the anemic processes observed at M0 (at least in cats with mild or moderate anemia) and leukocyte counts, including a more favorable CD4+/CD8+ ratio. An increase in the serum gammaglobulin concentration was seen in 80% of the cats. Despite observing an obvious favorable progress in the clinical, biopathological, and CD4+/CD8+ values during treatment, almost invariably all the parameters analyzed worsened after treatment discontinuation (M10), which suggests that the interferon-α protocol should be either extended or include additional cycles for a long-lasting benefit in FeLV+ and FIV+ cats.

Effect of biphasic temperature regime on therapeutic recombinant protein production in the green alga Chlamydomonas reinhardtii

Microalgae are increasingly being considered for recombinant protein production because of low cultivation costs, absence of endotoxins and insusceptibility to human infectious agents. Despite these advantages, the yield of recombinant protein produced in microalgae is still low compared to more established expression systems and optimization at the genetic and cultivation levels is required for this new system to be economically viable. This study investigates the effect of biphasic temperature regimes on the yield of recombinant human interferon alpha 2a (IFN-α2a), a therapeutic protein known for its anti-cancer and anti-viral properties, produced by the model green alga Chlamydomonas reinhardtii (Cr.IFN-α2a). Biphasic growth is commonly employed to increase recombinant protein production in mammalian cell lines used for commercial production of therapeutic proteins, with a lowering of the temperature resulting in higher yields. In this study, lowering the temperature from 25 °C to 15 °C in mid-exponential growth phase increased the accumulation of Cr.IFN-α2a by 3.3-fold while it slowed down the growth of the three C. reinhardtii transgenic lines tested. In contrast, a rise of temperature from 25 °C to 35 °C accelerated cell growth, while negatively impacting the production of Cr.IFN-α2a. After a two-step chromatography purification, the Cr.IFN-α2a produced was estimated to be 53% pure with a yield of 90 μg/L of culture. The amino acid sequence of Cr.IFN-α2a was confirmed by mass spectrometry. However, the anti-viral activity of Cr.IFN-α2a was found to be 10 times lower than the human IFN-α2a standard produced using E. coli when challenged in a cytopathic effect (CPE) assay, likely due to the formation of aggregates. While the molecular mechanisms driving the accumulation of Cr.IFN-α2a at lower temperature remains unclear, our results support that reducing the temperature at the peak of expression is a valid strategy to increase the yield of recombinant Cr.IFN-α2a in C. reinhardtii.

Еxternal forms of recombinant interferon alfa-2b - ointment and gel in the treatment of acute respiratory viral infections and influenza in children

The review article presents the results of numerous studies on the effectiveness and safety of the newly developed forms of the human recombinant interferon alpha- 2b drug, ointment and gel, intended for the treatment and prevention of acute respiratory viral infections and influenza in children. Еxternal forms of the drug retain immunomodulatory, antiviral and antiproliferative effects. The dosage form of the preparation of interferon alpha-2b VIFERON® in the form of a gel or ointment allows to achieve a prolonged action due to good absorption, especially the gel, which increases its effectiveness, and also has a moisturizing effect and rarely causes allergies. All of the above properties justify the inclusion of the drug interferon alpha-2b VIFERON® in the form of a gel or ointment in the complex treatment of influenza and acute respiratory viral infections in children.

Aspects of a Methodological Approach to Determination of Interferon Alpha Specific Activity

Specific antiviral activity is one of the key indicators characterising pharmaceutical quality and pharmacological efficacy of interferon alpha products (IFN-α). Specific activity is determined using a bioassay measuring antiviral activity in cell culture. The aim of the study was to select the most appropriate conditions for in vitro determination of IFN-α product specific activity. Materials and methods: Vero, MDBK, Hep-2, and A-549 homologous and heterologous cell cultures, as well as vesicular stomatitis Indiana virus (VSV) and murine encephalomyocarditis (EMC) virus at a dose of 100 TCD50 /0.1 mL were used for determination of specific antiviral activity. The international reference standard of recombinant interferon alpha-2b activity (Interferon alpha 2b, human, rDNA, E. coli-derived, 2nd WHO International Standard, 1999, NIBSC Code No. 95/566) and human recombinant interferon alpha 2b in the form of solution (batch No. 040214, Pharmapark LLC, Russia) were used as IFN-α samples. Results: the analysis of the obtained data helped to determine: the combinations of cell lines and the indicator virus most sensitive to IFN-α; the optimal concentration of fetal serum in the medium, and the optimal time parameters; the preferred method of reporting test results. Conclusions: the following test conditions were found to be optimal: the MDBK/VSV and Hep-2/EMC combinations proved to be the most sensitive to IFN-α; the optimal period of interferon and cell culture incubation—24 hours; the optimal concentration of fetal bovine serum in the culture medium used for diluting interferon products—2–5%. The instrumental procedure is preferred for reporting the results of interferon antiviral activity determination, because it is up-to-date, reliable, accurate and time-efficient.

A prospective multicenter randomized controlled study on the efficacy and safety of pharyngeal spraying recombinant human interferon alpha 2b in the treatment of children with herpangina

Objective To evaluate the efficacy and safety of pharyngeal spraying recombinant human interfe-ron alpha 2b (rhIFNα2b) in the treatment of herpangina in children. Method s A prospective, multicenter, rando-mized, opened and controlled study was carried out in 11 hospitals in Anhui province from August 2018 to March 2019.According to the time of admission, 180 patients diagnosed as herpangina were prospectively and randomly divided into rhIFNα2b treatment group and Ribavirin control group.On the basis of giving both groups the heat-clearing, detoxifying and anti-infection treatment, the patients in treatment group received pharyngeal spraying rhIFNα2b 9 g/L saline solution[1 million IU/mL, 0.1 million IU/(0.1 mL·press)], and the patients in control group were treated by pharyngeal spraying Ribavirin (0.5 mg RBV/press, 150 press), 3 presses per time, 4 times per day, continuous administration for 5 days for both groups.Those who recovered in advance were no longer given medication.All patients were observed to fully recover.The clinical efficacy and the disappearing time of symptoms and signs between two groups were compared, and the safety of pharyngeal spraying rhIFNα2b for patients was evaluated. Result s All of the 180 patients completed the study, including 90 cases in the treatment group and 90 cases in the control group.There was no statistically significant difference in terms of gender, age, weight and course of illness before treatment between the two groups (all P>0.05), which had clinical comparability.The apparent efficiency of the treatment group [63.3% (57/90 cases)] was significantly higher than that in the control group [38.9% (35/90 cases)] and the difference was statistically significant(χ2=10.934, P=0.004); no significant difference in the total efficiency between the treatment group [96.7% (87/90 cases)]and the control group [92.2% (83/90 cases)]was observed (χ2=2.924, P=0.169). The duration of fever[(32.59±20.73) h vs.(45.72±26.96) h], hyperemia[(76.48±23.12) h vs.(92.44±24.31) h], herpes[(72.99±25.77) h vs.(85.09± 26.62) h], salivation[(45.44±24.96) h vs.(54.42±31.20) h] and anorexia[(62.70±23.99) h vs.(78.71±30.54) h] in the treatment group were significantly shorter than those in the control group, and the differences were statistically significant(all P 0.05). After treatment, the serum levels of TNF-α and IL-6 were(7.26±1.99) ng/L and (2.42±0.73) ng/L in the treatment group, which were significantly lower than those in the control group [(12.09±6.39) ng/L and (7.32±11.51) ng/L](all P 0.05). The comparison on positive rate of virus in pharyngeal swab between the treatment group [65.3% (32/49 cases) and 40.6% (13/32 cases) respectively] and the control group[66.7%(36/54 cases) and 41.0% (16/39 cases), respectively]before and after therapy showed no significant difference (all P>0.05). During the treatment, no serious adverse reactions were observed in the two groups.The incidence of adverse reactions was 1.1% (1/90 cases) in the treatment group and 5.6% (5/90 cases) in the control group.In addition, the serum hemoglobin level of children in the control group after treatment was significantly lower than that before treatment and that in the treatment group (all P<0.05). Conclusions Compared with pharyngeal spraying ribavirin, pharyngeal spraying rhIFNα2b can greatly improve the clinical efficiency, accelerate the disappearance of clinical symptoms and signs, and shorten the total course of disease, and is more safe and worthy of clinical application. Key words: Herpangina; Recombinant human interferon alpha 2b; Ribavirin; Pharyngeal spraying

Therapeutic responses to interferon-alpha in HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP): Differing clinical responses to leukocyte-derived human interferon-alpha versus recombinant interferon-alpha

Introduction: HTLV-1-associated myelopathy/tropical spastic paraparesis (HAM/TSP) is a progressive spinal cord syndrome seen in patients infected by human T-lymphotropic virus type 1 (HTLV-1). Initial infection by the virus is usually asymptomatic. As the disease progresses, a se

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.

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

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

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.