Human Leukocyte Interferon Research Articles

Clinical results of the using leucocyte interferon for the therapy of viral pathology of the oral mucous in somatic patients with combined dentistry pathology

Aim. To conduct a clinical test of the unguent form of human leukocyte interferon in somatic patients with relapsing of herpetic stomatitis.Material and methods. 98 somatic patients with viral pathology of the oral mucous at the age of 20 to 60 years and older were studied to identify the clinical efficiency of the leukocyte interferon. The order of the clinical studies was specified by a special program: preparations were decrypted, decryption was conducted after the analysis of the results of the clinical study. Preparation 1 – placebo (2% emulsion of lidocaine on vinilin), preparation 2 – interferon’s unguent, preparation 3 – 0.5% oxoline unguent. After active implementation of the complex treatment of patients with mucologic pathology were divided into 3 groups: 1st group included 31 patients who got a course of the treatment by preparation 1; 2nd - 34 persons, 2 have taken preparation; 3rd - 33 patients, 3 used preparation.Results. The most expressed clinical effect in the treatment of relapsing of herpetic stomatitis in somatic patients with combined dentistry pathology was shown by preparation 2, which had local anesthetic and anti-inflammation action. Indicators of the recovery of patients who got this preparation in contrast with patients of the third group decreased on average by 2 days, the 1st groups - by more than 1 day.Conclusion. Using oxolin’s unguent and preparation 1 in course of the treatment of herpetic stomatitis did not prevent the development of secondary elements in the oral mucous (24.0% and 44.0% accordingly) in most of the patients. Using preparation 2, new elements on oral mucous were observed only in 6.0% of cases, which shows it good antiviral effect.

Biomedical Text Link Prediction for Drug Discovery: A Case Study with COVID-19.

Link prediction in artificial intelligence is used to identify missing links or derive future relationships that can occur in complex networks. A link prediction model was developed using the complex heterogeneous biomedical knowledge graph, SemNet, to predict missing links in biomedical literature for drug discovery. A web application visualized knowledge graph embeddings and link prediction results using TransE, CompleX, and RotatE based methods. The link prediction model achieved up to 0.44 hits@10 on the entity prediction tasks. The recent outbreak of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), also known as COVID-19, served as a case study to demonstrate the efficacy of link prediction modeling for drug discovery. The link prediction algorithm guided identification and ranking of repurposed drug candidates for SARS-CoV-2 primarily by text mining biomedical literature from previous coronaviruses, including SARS and middle east respiratory syndrome (MERS). Repurposed drugs included potential primary SARS-CoV-2 treatment, adjunctive therapies, or therapeutics to treat side effects. The link prediction accuracy for nodes ranked highly for SARS coronavirus was 0.875 as calculated by human in the loop validation on existing COVID-19 specific data sets. Drug classes predicted as highly ranked include anti-inflammatory, nucleoside analogs, protease inhibitors, antimalarials, envelope proteins, and glycoproteins. Examples of highly ranked predicted links to SARS-CoV-2: human leukocyte interferon, recombinant interferon-gamma, cyclosporine, antiviral therapy, zidovudine, chloroquine, vaccination, methotrexate, artemisinin, alkaloids, glycyrrhizic acid, quinine, flavonoids, amprenavir, suramin, complement system proteins, fluoroquinolones, bone marrow transplantation, albuterol, ciprofloxacin, quinolone antibacterial agents, and hydroxymethylglutaryl-CoA reductase inhibitors. Approximately 40% of identified drugs were not previously connected to SARS, such as edetic acid or biotin. In summary, link prediction can effectively suggest repurposed drugs for emergent diseases.

Science at the Service of Public Health: Rafael Pérez Cristiá MD PhD General Director, Center for State Control of Medicines and Medical Devices

Cuba’s nascent biotechnology sector began making scientific breakthroughs in the 1980s, including the isolation of human leukocyte interferon alpha (1981) and the development of the world’s first safe, effective meningitis BC vaccine (1989). With positive results in hand and a growing R&D pipeline, the island nation established a national regula-tory authority (NRA) to implement and oversee best practices for all pharmaceuticals and medical devices, domestically produced and imported, used in the country’s universal health system. Founded in 1989, Cuba’s Center for State Control of Medicines and Medical Devices (CECMED) is the entity charged with regulating all phases of scientific innovation for health, from clinical trial design to postmarketing surveillance. Dr Rafael Perez Cristia, Distinguished Member of the Cuban Academy of Sciences, has been Director General of CECMED since 2000, overseeing regulation and control of unique and innovative biotechnology products and the concomitant evolution of the nation’s regulatory authority. Under his guidance, CECMED has regulated unique therapies, vaccines, and pharmaceutical products—some unavailable anywhere else in the world—aimed at improving population health both at home and abroad. Recognized internationally as one of the top 20 countries with a safe and reliable biotechnology industry and regulatory authority, Cuba is having a measurable impact on public health. In this exclusive interview, Dr Perez Cristia, explains how a small, resource-scarce country has rocketed into the global biotech elite—and how it intends to stay there.

Additive Effects of Water-Soluble Propolis (Greit 120) and Human Interferon Alfa (HuIFN-αN3) against Influenza Viruses in Vitro

Influenza virus affects the respiratory tract in humans causing a range of distinct manifestations including fever, nasal secretions, cough, headaches, muscle pain and pneumonia, which could become violent and severe. It was found that influenza A viruses remain resistant to amantadine and rimantadin with high level of oseltamvir resistance. Therefore, there is a need for constant improvement of drugs active against resistant influenza viruses. Propolis has anti-influenza activity both in vitro and in vivo. Human leukocyte interferon (HuIFN-αN3) is a multi- subtype protein that displays an antiviral activity against influenza virus. In this study we elucidated the anti-influenza activity of the mixes of water-soluble propolis (WSP) (Greit 120) and HuIFN-αN3 at different ratios. Greit 120 polyphenols, total phenol acids and bioflavonoid were characterized by HPLC-UV-ESI- MS504971 and HuIFN-αN3 by reverse-phase high- performance liquid chromatography (RP-HPLC). Influenza A and B viruses were separately added to the LLC-MK2 cells treated with WSP (Greit 120) and HuIFN-αN3 alone or in proportions 1:1, 1:2 and 2:1. Plates were incubated and cytopathic effect was determined. The best results (ID50) were obtained with the mix of 10% WSP and HuIFN-αN3 in proportion 1:2, showing ID50 at 12 ± 0.2 μg/mL and 19 ± 0.6 μg/mL for influenza A and B viruses, respectively. When comparing anti- influenza activity of WSP (Greit 120)/HuIFN-αN3 with that of ribavirin, it was found that 1:2 was the optimal ratio for WSP (Greit 120)/HuIFN-αN3 (0.5 and 0.6 for influenza A and B, respectively). This new formulation of WSP (Greit 120) and HuIFN-αN3, showing better anti-Influenza activity, will definitely improve its application in flu infections.

Science at the Service of Public Health: Rafael Pérez Cristiá MD PhD Center for State Control of Medicines and Medical Devices.

Cuba's nascent biotechnology sector began making scientific breakthroughs in the 1980s, including the isolation of human leukocyte interferon alpha (1981) and the development of the world's first safe, effective meningitis BC vaccine (1989). With positive results in hand and a growing R&D pipeline, the island nation established a national regulatory authority (NRA) to implement and oversee best practices for all pharmaceuticals and medical devices, domestically produced and imported, used in the country's universal health system. Founded in 1989, Cuba's Center for State Control of Medicines and Medical Devices (CECMED) is the entity charged with regulating all phases of scientific innovation for health, from clinical trial design to postmarketing surveillance. Dr Rafael Pérez Cristiá, Distinguished Member of the Cuban Academy of Sciences, has been Director General of CECMED since 2000, overseeing regulation and control of unique and innovative biotechnology products and the concomitant evolution of the nation's regulatory authority. Under his guidance, CECMED has regulated unique therapies, vaccines, and pharmaceutical products-some unavailable anywhere else in the world-aimed at improving population health both at home and abroad. Recognized internationally as one of the top 20 countries with a safe and reliable biotechnology industry and regulatory authority, Cuba is having a measurable impact on public health. In this exclusive interview, Dr Pérez Cristiá, explains how a small, resource-scarce country has rocketed into the global biotech elite-and how it intends to stay there.

The ABCs of Clinical Trials in Cuba.

In 1981, Cuba's biotechnology sector was born when Cuban scientists, trained by Finnish colleagues, successfully isolated large quantities of human leukocyte interferon alpha. Within a decade and following a $1-billion dollar investment by the Cuban government, 52 scientific research institutions were established in and around Havana to develop vaccines, therapies and other medical applications to meet demands of the national health system.

The ABCs of Clinical Trials in Cuba.

In 1981, Cuba's biotechnology sector was born when Cuban scientists, trained by Finnish colleagues, successfully isolated large quantities of human leukocyte interferon alpha. Within a decade and following a $1-billion dollar investment by the Cuban government, 52 scientific research institutions were established in and around Havana to develop vaccines, therapies and other medical applications to meet demands of the national health system.

135: Mycobacterium tuberculosis strains induce differential cytokine production and gene expression patterns in pulmonary alveolar epithelial cells

Aims To determine whether cytokine production and gene expression levels vary in epithelial cells infected by virulent strains of Mycobacterium tuberculosis . Methods Supernatants were collected and RNA extracted from A549 epithelial cells infected with the KZN, Beijing, F11, F28, Unique and H37Rv strains of M. tuberculosis . Cytokines were quantified using the Bio-Plex Pro Human Cytokine 27-Plex assay (BioRad) and the Bio-Plex 200 system. Changes in gene expression were determined by whole transcriptome analysis using the Illumina HiSeq 2000 platform. The 50 bp reads were mapped to the human genome (hg19) using Tophat (2.0.10). Differential expression was quantified using Cufflinks (2.1.0) and a log fold change cutoff of 2. Results Twenty-three analytes were detected. All strains, except the F28 induced an increase in the production of 18, and a decrease in 5 cytokines/chemokines at 24, 48 and 72 h post-infection, compared to the uninfected control. However, F28 induced an increased production of all 23 cytokines/chemokines at 48 and 72 h. A gradual increase in anti-, pro-inflammatory cytokines and chemokines that peaked mostly at either 48 or 72 h, was induced in a strain specific manner. Changes in host gene expression were observed in surface receptors, interleukins, chemokines, transcriptional factors, human leukocyte antigen and interferon associated proteins. High fold changes were observed for HLA (B, C, F and H) genes, TLR5 and NOD2 receptors. The Unique strain induced the lowest fold changes while the F15/LAM4/KZN and F11 strains induced the highest fold changes in most genes. Hierarchical clustering showed that the Unique strain clustered separately from the highly virulent ones. Conclusion Clinical strains of M. tuberculosis of different genotypes induce specific patterns of in vitro cytokine production and immune gene expression in pulmonary epithelial cells. Furthermore, our results support findings that lung epithelial cells are involved in antigen processing and presentation.

Human Leukocyte Antigen Genes and Interferon Beta Preparations Influence Risk of Developing Neutralizing Anti-Drug Antibodies in Multiple Sclerosis

A significant proportion of patients with multiple sclerosis who receive interferon beta (IFNβ) therapy develop neutralizing antibodies (NAbs) that reduce drug efficacy. To investigate if HLA class I and II alleles are associated with development of NAbs against IFNβ we analyzed whether NAb status and development of NAb titers high enough to be biologically relevant (>150 tenfold reduction units/ml) correlated with the HLA allele group carriage in a cohort of 903 Swedish patients with multiple sclerosis treated with either intramuscular IFNβ-1a, subcutaneous IFNβ-1a or subcutaneous IFNβ-1b. Carriage of HLA-DRB1*15 was associated with increased risk of developing NAbs and high NAb titers. After stratification based on type of IFNβ preparation, HLA-DRB1*15 carriage was observed to increase the risk of developing NAbs as well as high NAb titers against both subcutaneous and intramuscular IFNβ-1a. Furthermore, in patients receiving subcutaneous IFNβ-1a carriage of HLA-DQA1*05 decreased the risk for high NAb titers. In IFNβ-1b treated patients, HLA-DRB1*04 increased the risk of developing high NAb titers, and in a subgroup analysis of DRB1*04 alleles the risk for NAbs was increased in DRB1*04:01 carriers. In conclusion, there is a preparation-specific genetically determined risk to develop NAbs against IFNβ high enough to be clinically relevant in treatment decisions for patients with multiple sclerosis if confirmed in future studies. However, choice of IFNβ preparation still remains the single most significant determinant for the risk of developing NAbs.

Interferon for Hepatitis C: Where it has been and where it is Going

ImmunotherapyVol. 5, No. 7 EditorialInterferon for hepatitis C: where it has been and where it is goingRonald L KoretzRonald L KoretzDepartment of Medicine, Olive View – UCLA Medical Center, 14445 Olive View Drive, Sylmar, CA 91340, USA. Search for more papers by this authorEmail the corresponding author at rkoretz@msn.comPublished Online:5 Jul 2013https://doi.org/10.2217/imt.13.55AboutSectionsView ArticleView Full TextPDF/EPUB ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareShare onFacebookTwitterLinkedInReddit View articleKeywords: hepatitis Cinterferonsurrogate end pointtreatmentReferences1 Greenberg HB, Pollard RB, Lutwick LI, Gregory PB, Robinson WS, Merigan TC. Effect of human leucocyte interferon on hepatitis B infection in patients with chronic active hepatitis. N. Engl. J. 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Med.368,45–53 (2013).Crossref, Medline, CAS, Google ScholarFiguresReferencesRelatedDetailsCited ByAn injectable thermosensitive hydrogel/nanomicelles composite for local chemo-immunotherapy in mouse model of melanoma11 May 2022 | Journal of Biomaterials Applications, Vol. 37, No. 3 Vol. 5, No. 7 Follow us on social media for the latest updates Metrics Downloaded 43 times History Published online 5 July 2013 Published in print July 2013 Information© Future Medicine LtdKeywordshepatitis Cinterferonsurrogate end pointtreatmentFinancial & competing interests disclosureThe author has no relevant affiliations or financial involvement with any organization or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.No writing assistance was utilized in the production of this manuscript.PDF download

Nanofizyka ta antyvirusna terapiya

A new mechanism of interaction between viruses and nanoparticles is proposed. The mechanism is based on the local-field enhancement effect inherent only in nano-objects and can manifest itself in nanoparticle–virus systems. The basic idea consists in vacuum fluctuations that are always present in any physical system. This mechanism is universal and does not depend on the details of nanoparticle and virus structures, which was confirmed by numerous experiments carried out by us and in other scientific groups. A new method of purification of biofluids from nano-objects such as nanoparticles and viruses is also discussed. The method is based on a selective interaction between nano-objects and either a nanostructured surface, along which a surface plasmon-polariton propagates, or a system of nanothreads, on which a local plasmon-polariton is excited. On the basis of the method proposed for weakening the virus activity due to the action of a suspension of nanoparticles, a new effective way for the production of human leukocytic interferon has been developed and verified experimentally.

Abstract C110: Mutation and chemosensitivity profile of a panel of 25 patient-derived melanoma xenografts.

Abstract Melanomas account for a notable proportion of cancers worldwide. The heterogeneity and biological characteristics of melanomas present unique therapeutic challenges. Recently a number of somatic mutations were determined, which influence the prognosis and thus therapeutic needs markedly. Here we present a panel of patient-derived melanoma xenografts, established by Oncotest from primary patient material. From 80 patient explants, growth in serial passage was observed in 43% of the cases. Molecular profiling included mutational analysis for BRAF, PI3Kalpha, KRAS, NRAS, TP53, CTNNB1, MET and JAK3 using OncoCarta™ and exon sequencing. The chemosensitivity profile was determined in vitro using 2D and 3D culture assays. Clinically relevant BRAF mutations were the most prevalent mutation and were found in almost two thirds of the melanoma models. The second most common gene mutated was NRAS, with about one quarter of the models harbouring activating mutations of NRAS. PI3Kalpha mutations were rare, and for KRAS no mutations were detected. Most of the melanomas showed multiple mutations of the investigated genes. Chemosensitivity profiling included cytotoxic (e.g. cisplatin, carboplatin, vinblastine, vincristine, ifosfamide, paclitaxel, and docetaxel), as well as targeted drugs (e.g. sorafenib and PLX-4720). The compounds showed diverse patterns of selectivity and potency: Differential activity was determined e.g. for vinblastine, vincristine, paclitaxel, and docetaxel, but also for the BRAF inhibitor PLX-4720. PLX-4720 was most active in melanoma models carrying the BRAF mutation as determined in monolayer and soft agar assays. Activity of vincristine revealed a correlation of sensitivity with a BRAF wild type genotype and resistance with BRAF mutation. The latter result was also described recently for melanoma patients with BRAF mutations, showing a diminished duration of response to treatment with vincristine, dacarbazine, bleomycin, lomustine, and human leukocyte interferon. A collection of 25 patient-derived melanoma models is presented. Their mutational status as well as their responsiveness towards many cytotoxic and targeted agents was analyzed. This extensive genotypic and phenotypic characterization makes them a valuable tool for hypothesis generation and testing. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2011 Nov 12-16; San Francisco, CA. Philadelphia (PA): AACR; Mol Cancer Ther 2011;10(11 Suppl):Abstract nr C110.

Regeneration of transgenic plants from hairy roots of Cichorium intybus L. var. Foliosum Hegi

Transgenic plants containing either ifn-α2b gene encoding human leukocytic interferon or esxA::fbpBΔTMD genes encoding Mycobacterium tuberculosis antigens ESAT6 and Ag85B were regenerated from hairy root cultures after transformation of chicory cotyledons (Cichorium intybus L. var. Foliosum Hegi) with a wild-type A. rhizogenes A4 strain. The direct shoot regeneration from transgenic roots without callus formation phase on growth regulator-free nutrient medium was demonstrated. The transgenes transfer and transcription in the plants were confirmed by the results of RT-PCR and PCR analyses.

Inhibition of Endogenous Interferon Beta by Neutralizing Antibodies Against Recombinant Interferon Beta

To determine if neutralizing antibodies (NAbs) against interferon beta from patients with multiple sclerosis (MS) cross-react with other type 1 interferons, especially endogenous interferon beta, and thus might impede the immune systems of affected patients. Masked serum samples from MS patients were challenged in vitro against recombinant interferon beta-1a and interferon beta-1b, as well as human leukocyte interferon and fibroblast interferon, the latter representing endogenous interferon. The neutralizing capacity of serum samples on these type 1 interferons was assessed using a luciferase reporter gene assay. Randomly selected samples were titrated to further delineate the cross-reactivity of antibodies. University medical center in Düsseldorf, Germany. We randomly selected 150 samples from interferon beta-treated MS patients who had previously been tested for the presence of binding antibodies and NAbs. Neutralization of interferon beta bioactivity and cross-reactivity of anti-interferon beta antibodies. Antibody-mediated neutralization of interferon beta bioactivity in vitro against recombinant interferon beta was observed in all serum samples that had previously tested positive for binding antibodies and NAbs. A neutralizing pattern comparable to that of recombinant interferon beta was observed when endogenous interferon was assessed, reflecting cross-reactivity of NAbs. No differences in neutralization between recombinant and endogenous interferon were observed with respect to the interferon beta preparation used for treatment. Furthermore, no neutralization of other type 1interferons by NAbs could be detected. A proportion of MS patients who are treated with recombinant interferon beta develop NAbs that also neutralize endogenous interferon. Because NAbs at high titers can persist for years, these antibodies may impede the immune system in affected MS patients regardless of their current treatment regimen.

Abstract A173: Profiling of targeted anticancer agents in a panel of 48 patient-derived xenografts using a clonogenic assay

Abstract Novel experimental and approved cancer therapeutics target proteins with key functions in mitosis regulation, cell survival and migration, protein turnover or angiogenesis. These targets include receptor tyrosine (Y) kinases like EGF-R, PDGF-R, VEGF-R, c-KIT and FLT-3, serine-threonine (S/T) specific kinases like B-raf and Aurora A/B (ARK), mitotic kinesins like Eg5 (KSP), the chaperone Hsp90 and traditional targets, namely topoisomerase I / II and ß-tubulin. At Oncotest, a unique profiling screen for anticancer agents using a clonogenic assay using a panel of 48 proprietary solid tumor xenografts is routinely done. The xenograft models in general were established directly from patient tumor material, representing all major tumor histotypes (non-small cell lung cancer / NSCLC, pancreatic, prostate, colon, gastric, breast, ovarian and renal cancer, melanoma and sarcoma) as well as niche tumors (pleuramesothelioma, bladder and head and neck cancer). After an initial profiling screen in the 48 tumor panel, follow-up testing in defined tumor histotypes are of high value for hypothesis generation including selection of a clinical development strategy. Here, activity profiles for inhibitors targeting EGF-R (Erlotinib), Aurora A/B (VX-680), multiple S/T and Y kinases (Sunitinib, Sorafenib), Eg5 (Ispinesib) and tububiln (Vincristine), Hsp90 (17-DMAG) and topoisomerase I (SN-38, active metabolite of Irinotecan) are discussed. Erlotinib displayed a diverse pattern of activity with a mean IC50 = 20.7µM. Selectivity was studied in an extended panel of NSCLC with tumors showing activated EGF-R being mainly most sensitive. The spectrum-kinase inhibitors Sunitinib and Sorafenib displayed weak selectivity with mean IC50 values of 7.9µM and 11.8µM, respectively. The Hsp90 inhibitor 17-DMAG, the topoisomerase I inhibitor SN-38 and Vincristine were highly potent with mean IC50 values of 28nM, 45nM, and 100nM, respectively. For 17-DMAG and SN-38 as examples, tumor xenograft models selected based on clonogenic assay data responded very well in in vivo efficacy tests. Further evaluation of Vincristine in an extended melanoma panel revealed a strong correlation of sensitivity with a B-raf wild type genotype and resistance with the B-raf V600E mutation. This result was described recently for melanoma patients with B-raf mutations, showing a diminished duration of response to treatment with Dacarbazine, Vincristine, Bleomycin, Lomustine, and human leukocyte interferon. In conclusion, the Oncotest clonogenic assay by retaining important characteristics of the original patient tumor is of high value for profiling of traditional cytotoxic as well as new targeted cancer agents. By revealing diverse activity and resistance patterns, molecular characterization data including gene mutations and expression are guiding the selection of tumor entities and patients likely to respond to therapy. Citation Information: Mol Cancer Ther 2009;8(12 Suppl):A173.

Neurotoxic and other side effects of high-dose interferon in amyotrophic lateral sclerosis.

6 patients with amyotrophic lateral sclerosis were treated with intravenous infusion of 100-200 million IU per day of human leukocyte interferon. Side effects of treatment included fever, chills, malaise, nausea, marked leukopenia, mild anemia, and thrombocytopenia. Tiredness, confusion, papilledema, and overall signs of acute encephalitis were observed. Tendon reflexes and muscle force decreased. EEG activity was slowed, and evoked potentials showed significant slowing of conduction times. Neuropsychological tests revealed congitive dysfunction. The syndrome of inappropriate antidiuretic hormone secretion developed in all patients. All side effects were reversible with cessation of interferon treatment.

Memory and psychomotor impairment following high-dose interferon treatment in amyotrophic lateral sclerosis.

Patients with amyotrophic lateral sclerosis were treated with high-dose intravenous infusion of human leukocyte interferon for six days. Neuropsychological examinations were carried out before, during and after the treatment. Marked reversible dysfunction was detected in immediate memory functions, coordination of hand movements, and drawing. Motor perseveration, micrographia, and slowing of behaviour were also observed. Changes appeared four to 12 days after start of treatment, with the peak on days six to eight. Recovery was almost complete by day 15. Intellectual ability, as measured by three WAIS subtests, praxis of hand movements, visuognostic functions, speech, reading, writing, and calculation remained essentially unaffected. The profile of the neuropsychological deficits observed, the absence of defects typical of focal posterior cortical lesions, the simultaneously slowed electroencephalographic activity with frontal accentuation, and the increased central conduction times of brain stem auditory evoked potentials suggest frontobasal involvement.

Autoantibodies to crude human leucocyte interferon (IFN), native human IFN, recombinant human IFN-alpha 2b and human IFN-gamma in healthy blood donors

Recently, naturally occurring antibodies to IFN-alpha were discovered in a few systemic lupus erythematosus (SLE) and cancer patients; however, in most patients monitored for anti-IFN antibodies before treatment, no antibodies were found. In an attempt to explain the 'IFN-blocking effect' that we observed in all serum samples we investigated 200 sera from healthy blood donors. We isolated the globulin fraction, and used rabbit anti-human IgG and IgM columns, protein A columns and T-gel affinity chromatography to isolate human IgG and IgM. All sample fractions were tested in a biological IFN neutralization assay by means of a sensitive MTT-assay. We found that normal human serum contained autoantibodies to crude human leucocyte IFN, native human fibroblast IFN, recombinant human leucocyte IFN-alpha 2b and recombinant human IFN-gamma, and that these naturally occurring antibodies were biologically active immunoglobulins of IgG and IgM type. These anti-IFN antibodies were also present in purified human normal immunoglobulin pools. We conclude that all humans have naturally occurring anti-interferon antibodies in their serum, and it is a tempting theory that human cytokines and lymphokines are, at least partly, regulated by immunoglobulins.

The purification and characterization of alpha interferons and related cytokine receptors—A personal account

In 1976–1977, I adapted reversed-phase HPLC (RP-HPLC) to peptide and protein purification, starting with pituitary proteins and continuing with the first successful purification to homogeneity of human leukocyte interferon (IFN-α). Using this technology, I isolated and characterized 6–8 different leukocyte interferon subtypes, which were later identified as products of the IFN-α gene family. Since then, RP-HPLC became a standard procedure for isolation and analysis of proteins. The successful purification of IFN-α led to the development of Roferon-A™, a drug used for the treatment of hairy cell leukemia, hepatitis C and a variety of other diseases. Later studies with my colleagues in Israel and abroad led to isolation and discovery of several cytokine receptors and binding proteins, including those of Type I IFNs, TNF and IL-18. The use of HPLC was indispensable in most of these studies.

Toll-like Receptor-dependent and -independent Viperin Gene Expression and Counter-regulation by PRDI-binding Factor-1/BLIMP1

Here we identify Viperin as a highly inducible gene in response to lipopolysaccharide (LPS), double-stranded RNA (poly(I-C)) or Sendai virus (SV). The only known function of Viperin relates to its ability to inhibit human Cytomegalovirus replication. Very little data are available on the regulation of this gene. In silico analysis of the promoter identified two interferon (IFN)-stimulated response elements (ISRE), which in other genes bind IRF3 or the IFN-stimulated gene factor-3 (ISGF3) complex. LPS and poly(I-C) induce very high levels of Viperin in wild type cells but not in cells deficient in TRIF, TBK1, IRF3, or the type I IFNalpha/betaR. SV-induced Viperin gene expression was mediated independently of Toll-like receptor (TLR) signaling by retinoic acid-inducible gene (RIG-I) and the downstream adapter, mitochondrial anti-viral signaling (MAVS). Virus-induced Viperin expression was not attenuated in macrophages deficient in either TBK1 or IKKepsilon alone. Moreover, IRF3-deficient, but not IFNalpha/betaR deficient, macrophages still induced Viperin in response to SV. Promoter reporter studies combined with DNA immunoprecipitation assays identified the ISGF3 complex as the key regulator of Viperin gene expression. Moreover, positive regulatory domain I-binding factor 1 (PRDI-BF1, also called BLIMP1) binds the ISRE sites and competes with ISGF3 binding in a virus inducible manner to inhibit Viperin transcription. Collectively, these studies identify Viperin as a tightly regulated ISGF3 target gene, which is counter-regulated by PRDI-BF1.