Recombinant Interferon Research Articles

INFLUENCE OF INTERLEUKIN-2 AND INTERFERON-α ON LYMPHOCYTE AGGREGATION AND LYMPHOCYTE-PLATELET CLUSTER FORMATION

The aim of the research was to study the direct and platelet-mediated intercellular adhesion of blood-derived lymphocytes, as well as the influence of interleukin-2 and interferon-α on it.Materials and methods. Whole blood samples from 34 apparently healthy individuals were collected using vacuum tubes containing sodium citrate (3,8%). A suspension of lymphocytes and platelets was isolated on a Ficoll-Urografin gradient. Light microscopy was used to determine the percentage of lymphocyte-platelet aggregates. The effect of cytokines was studied by adding human recombinant interleukin-2 and interferon-α to whole blood, and incubating for 4 hours in a thermostat at 37 °C. After incubation the necessary parameters were counted using the method described above. The results were expressed as mean values and standard deviations (± SD). Statistical processing of the data was performed using the Mann-Whitney U-test and the Kolmogorov criterion (Statistica 10), with differences considered significant at p < 0.05.Results. The study showed that in addition to lymphocyte-platelet aggregates (11 ± 3.6%), the total pool of lymphocytes also contained intercellular aggregates of lymphocytes (3 ± 3,8 per 100 cells) and lymphocyteplatelet clusters (2 ± 0,6 per 100 cells). It was found that the addition of interleukin-2 (IL-2) led to an increase in lymphocyte-platelet aggregates (LPA) and lymphocyte-platelet clusters by 1,8 times (p < 0,001) and 3,3 times (p < 0,001), respectively, compared to the control group. In contrast, incubation of blood samples with interferon-α (IFN-α) led to a decrease in the number of LPA (by 5.5 times compared to the control, p < 0,001) and almost prevented the ability of lymphocytes and platelets to form clusters. The presence of the abovementioned cytokines in the incubated blood did not affect the ability of lymphocytes to form aggregates with each other.Сonclusion. It was found that IL-2 increases the ability of lymphocytes and platelets to form clusters, while IFN-α significantly reduces this ability and has an inhibitory effect on the ability of these cells to form LPA. In our opinion, it is important that the effects of these cytokines were manifested only upon contact of lymphocytes with platelets.

Yuri Anatolyevich Ovchinnikov and I. The Big is Seen from a Distance

I have talked about Yuri Anatolyevich in many popular articles. We have more than 30 publications in common. In particular, more than 10 are on RNA polymerase:, 5 are on ATPase, 1 (out of 3) is devoted to photoaffinity modification of the active centers of RNA polymerase; 18 articles and patents on interferons, 10 of which involve Yuri Anatolyevich: 6 of the most significant, 7 articles and patents (3 of which involve Yuri Anatolyevich) are devoted to the problem of a vaccine against the Hepatitis A virus. I was part of the team that received the USSR State Prize in 1981 for work on RNA polymerase, headed by Yuri Anatolyevich. It should also be noted that with the active assistance of Yu. A. Ovchinnikov, industrial production of interferon was organized, which I called Reaferon (recombinant alpha interferon), and I, together with other participants in this difficult work, was awarded the Lenin Prize. Reaferon is sold in pharmacies to this day and will be used for a very long time. I have provided this data so that it is clear how close our cooperation was.

Brief Communication: Combination of an MIP3α-Antigen Fusion Therapeutic DNA Vaccine With Treatments of IFNα and 5-Aza-2'Deoxycytidine Enhances Activated Effector CD8+ T Cells Expressing CD11c in the B16F10 Melanoma Model.

Previous studies in the B16F10 mouse melanoma model have demonstrated that combining a DNA vaccine comprised of regions of gp100 and tyrosinase-related protein 2 fused to macrophage-inflammatory protein 3-alpha (MIP3α) with recombinant interferon alpha (IFN) and 5-Aza-2'-deoxycytidine (5Aza) treatments resulted in significantly greater antitumor activity and immunogenicity in the tumor microenvironment (TME). This brief report details that the combination of vaccine with treatments IFN and 5Aza results in an increase in the TME of a distinct CD11c+ CD8+ T-cell population. This cell population correlates with tumor size, is primarily comprised of effector or effector memory T cells, and has a more robust response to ex vivo stimulation as compared with CD11c- CD8+ T cells. In conclusion, this combination therapy results in a greater presence of highly active effector CD8+ T cells expressing CD11c in the TME, which are likely primary contributors to treatment efficacy.

Fluorescence-Based Study of Oligonucleotide Interactions With Recombinant Proteins: Insulin, Interferon α2-β, Somatotropin, and Their Receptors

Background. Oligonucleotides (OLNs) can participate in a wide range of protein-ligand interactions and perform numerous cellular functions by forming structures that enable specific interactions with DNA, RNA, and proteins, what is crucial for many biological processes. Advances in understanding these interactions could lead to the development of new technologies for treating various diseases. However, the mechanism of interaction between proteins and OLNs is complex and still requires detailed study. More research is needed to fully elucidate this process and enhance our understanding of these biomolecular interactions. Objective. The aim of this study was to synthesize, purify, and investigate the interaction of OLNs with recombinant signaling proteins interferon α2-β and insulin with their receptors and somatropin by assessing binding strength using fluorescence spectroscopy. Methods. The interactions were analyzed using the Stern–Volmer equation in both general and modified forms, as well as the Hill equation. OLNs were synthesized via the solid-phase phosphoramidite method, purified through solid-phase extraction, and subsequently verified with a spectrophotometer. Results. Fluorometric titration revealed that OLNs bind to proteins within the medium affinity range, forming non-fluorescent complexes, with the most active interactions observed with shorter OLN. Positive cooperative binding of interferon to G20 and T20, and negative cooperative binding of insulin to C20 and A20, were identified. Additionally, negative cooperative binding of somatropin to C20 was observed. Conclusions. The study demonstrated the interaction between OLNs and recombinant signaling proteins and receptors through various binding mechanisms, which could potentially affect their conformation and biolo­gical activity. These findings have implications for the therapeutic use of OLNs in the context of signaling proteins and receptors.

Study on the Recombinant Human Interferon α1b, α2b, and Gamma Transient Expression and in Vitro Activities in Tobacco.

Interferons (IFNs) are universally acknowledged for their pivotal role in antiviral and anticancer responses. Thus, the primary aim of our study was to explore the expressions of IFN-α1b, α2b, and gamma in tobacco leaves via agrobacterium-mediated transient transformation and investigate their possible activities. Briefly, fusion with green fluorescent protein tags aided in detecting the expressed IFN proteins in the foliar tissues. The genetic constructs encoding these fusion proteins were inserted into the MagnICON plant transient expression vector, followed by transformation into the Agrobacterium strain GV3101. The transformed bacteria were then used to infiltrate tobacco leaves. After post-infiltration, protein expression was confirmed within 72 h via sodium dodecyl sulfate polyacrylamide gel electrophoresis, and the fusion proteins were subsequently purified using high-performance liquid chromatography for identification. Both the antiviral and anticancer potencies of these IFN fusion proteins were evaluated using the WISH/VSV (WISH cells/Vesicular stomatitis virus) microneutralization and MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays, respectively. Results indicated robust expression of the targeted IFN genes in plant tissues and significant biological activities against pathogens and cancer cells. Consequently, this study substantiated the viability of producing these therapeutic proteins in plants, potentially revolutionizing the manufacture of interferons biologically.

Evaluation of the pharmacokinetics of an experimental combined form of IFNα and IFNγ for intranasal use

Polypous rhinosinusitis is a chronic inflammatory disease of the nasal mucosa and paranasal sinuses. Recombinant interferon has antiproliferative activity and corrects the deficiency of endogenous regulatory molecules, which allows us to consider this class of immunotropic drugs as a promising component of conservative immunotherapy for polyposis rhinosinusitis. Purpose of the study: To select the optimal composition of the experimental composition of interferons and evaluate their pharmacokinetic parameters in laboratory animals.For laboratory evaluation of the effectiveness of the auxiliary components of the proposed composition in creating a local effect, Wistar rats in the amount of 30 animals were selected, to which the proposed form of the drug (IFN1) was administered once intranasally. The control group included 30 animals that were alternately administered intranasally with a similar dose of IFNα2b and IFNγ dissolved in water for injection (IFN2). Quantitative determination of interferon concentration in blood samples was carried out using the enzyme immunoassay method. To assess the dependence of changes in concentrations in the blood of animals on the time elapsed after their administration, standard pharmacokinetic models were used. Next, the integral from the initial moment of time to infinity was determined, which corresponded to the area under the pharmacokinetic curve and made it possible to calculate a number of pharmacokinetic characteristics.The following indicators were obtained: 1) Area under the pharmacokinetic curve (AUCt) ng/ml/min – IFN1 = 683.0; IFN2 = 1707.7. 2) Absorption constant (Kp) – IFN1 = 0.13096; IFN2 = 0.03836. 3) Suction constant (Kel) – IFN1 = 0.00177; IFN2 = 0.00317. 4) Clearance (Cl) ml/min – IFN1 = 129.35; IFN2 = 51.73.Based on the differences in the values of pharmacokinetic parameters (AUCt) for the IFN1 and IFN2 preparations, it can be concluded that the use of a composition based on chitosan, succinic acid and DMSO leads to a pronounced retention of interferon in the nasal mucosa, which provides a pronounced local therapeutic effect and allows for fewer systemic adverse reactions. This composition is suitable for further clinical studies of the effectiveness of immunotherapy for polyposis rhinosinusitis.

Combination of a MIP3α-antigen fusion therapeutic DNA vaccine with treatments of IFNα and 5-Aza-2'Deoxycytidine enhances activated effector CD8+ T cells expressing CD11c in the B16F10 melanoma model.

Previous studies in the B16F10 mouse melanoma model have demonstrated that combining a DNA vaccine comprised of regions of gp100 and tyrosinase-related protein 2 fused to Macrophage-inflammatory protein 3-alpha (MIP3α) with recombinant Interferon alpha (IFN) and 5-Aza-2'-Deoxycytidine (5Aza) treatments resulted in significantly greater anti-tumor activity and immunogenicity in the tumor microenvironment (TME). This brief report details that the combination of vaccine with treatments IFN and 5Aza results in both the upregulation of genes expressing CD11c-interacting proteins and an increase in the TME of a distinct CD11c+ CD8+ T cell population. This cell population correlates with tumor size, is primarily comprised of effector or effector memory T cells, and has a more robust response to ex vivo stimulation as compared to CD11c- CD8+ T cells as measured by surface activation markers 4-1BB (CD137) and KLRG1 (Killer cell lectin-like receptor G1) and intracellular IFNγ production. In conclusion, this combination therapy results in greater presence of highly active effector CD8+ T-cells expressing CD11c in the TME that correlate with and are likely primary contributors to treatment efficacy.

Combination of a MIP3α-antigen fusion therapeutic DNA vaccine with treatments of IFNα and 5-Aza-2'Deoxycytidine enhances activated effector CD8+ T cells expressing CD11c in the B16F10 melanoma model.

Previous studies in the B16F10 mouse melanoma model have demonstrated that combining a DNA vaccine comprised of regions of gp100 and tyrosinase-related protein 2 fused to Macrophage-inflammatory protein 3-alpha (MIP3α) with recombinant Interferon alpha (IFN) and 5-Aza-2'-Deoxycytidine (5Aza) treatments resulted in significantly greater anti-tumor activity and immunogenicity in the tumor microenvironment (TME). This brief report details that the combination of vaccine with treatments IFN and 5Aza results in both the upregulation of genes expressing CD11c-interacting proteins and an increase in the TME of a distinct CD11c+ CD8+ T cell population. This cell population correlates with tumor size, is primarily comprised of effector or effector memory T cells, and has a more robust response to ex vivo stimulation as compared to CD11c- CD8+ T cells as measured by surface activation markers 4-1BB (CD137) and KLRG1 (Killer cell lectin-like receptor G1) and intracellular IFNγ production. In conclusion, this combination therapy results in greater presence of highly active effector CD8+ T-cells expressing CD11c in the TME that correlate with and are likely primary contributors to treatment efficacy.

Effects of ganciclovir combined with recombinant human interferon-α on clinical efficacy and immune function in children with infectious mononucleosis.

To evaluate the effects of ganciclovir combined with recombinant human interferon on clinical efficacy and immune function of children with infectious mononucleosis(IM). This was a retrospective study. Children (n=120) with IM hospitalized in Beijing Children's Hospital Affiliated to Capital Medical University Baoding Hospital from January 2020 to January 2022 were selected and randomly divided into study group and control group((n=60). Patients in the control group were treated with ganciclovir by intravenous infusion, and patients in the study group were given ganciclovir+recombinant human interferon-α1b. The time for eliminating clinical symptoms, the levels of inflammatory cytokines, immune function condition and T-lymphocyte subsets between the two groups were compared and analyzed. After treatment, the time for body temperature returned to normal, time for recovery from cervical lymphadenopathy, time for recovery from hepatosplenomegaly and time for disappearance of angina and oral mucosal congestion in the study group were significantly shorter than those in the control group(p= 0.00); after treatment, the levels of TNF-a and IL-6 in the study group were significantly lower than those in the control group; the indexes of CD3+ and CD8+ in the study group were significantly lower than those in the control group; after treatment, the levels of CD4+ and CD4+/CD8+ in the study group were significantly higher than those in the control group. Ranciclovir combined with recombinant human interferon-α1b, rapid improvements of clinical symptoms, significantly decreased inflammatory cytokines, improved T-lymphocyte function and no significant increase in adverse drug reactions were found in children with IM.

Recombinant porcine interferon delta 8 inhibits swine acute diarrhoea syndrome coronavirus infection in vitro and in vivo

Swine acute diarrhoea syndrome coronavirus (SADS-CoV), which originates from zoonotic transmission of bat coronaviruses in the HKU2 lineage, causes severe illness in pigs and carries a high risk of spreading to humans. At present, there are no licenced therapeutics for the treatment of SADS-CoV. In this study, we examined the effectiveness of recombinant porcine interferon delta 8 (IFN-δ8) against SADS-CoV both in vitro and in vivo. In vitro experiments showed that IFN-δ8 inhibited SADS-CoV proliferation in a concentration-dependent manner, with complete inhibition occurring at a concentration of 5 μg/mL. In vivo experiments demonstrated that two 50 μg/kg doses of IFN-δ8 injected intraperitoneally protected piglets against lethal challenge, blocked viral shedding, attenuated intestinal damage, and decreased the viral load in the jejunum and ileum. Further findings suggested that IFN-δ8 inhibited SADS-CoV infection by increasing the expression of IFN-stimulated genes. These results indicate that IFN-δ8 shows promise as a biological macromolecule drug against SADS-CoV infection.

Preparation and evaluation of microencapsulated delivery system of recombinant interferon alpha protein from rainbow trout

Diseases caused by viruses pose a significant risk to the health of aquatic animals, for which there are presently no efficacious remedies. Interferon (IFN) serving as an antiviral agent, is frequently employed in clinical settings. Due to the unique living conditions of aquatic animals, traditional injection of interferon is cumbersome, time-consuming and labor-intensive. This study aimed to prepare IFN microcapsules through emulsion technique by using resistant starch (RS) and carboxymethyl chitosan (CMCS). Optimization was achieved using the Box-Behnken design (BBD) response surface technique, followed by the creation of microcapsules through emulsification. With RS at a concentration of 1.27 %, a water‑oxygen ratio of 3.3:7.4, CaCl2 at 13.67 %, CMCS at 1.04 %, the rate of encapsulation can escalate to 80.92 %. Rainbow trout infected with Infectious hematopoietic necrosis virus (IHNV) and common carp infected with Spring vireemia (SVCV) exhibited a relative survival rate (RPS) of 65 % and 60 % after treated with IFN microcapsules, respectively. Moreover, the microcapsules effectively reduced the serum AST levels and enhanced the expression of IFNα, IRF3, ISG15, MX1, PKR and Viperin in IHNV-infected rainbow trout and SVCV-infected carp. In conclusion, this integrated IFN microcapsule showed potential as an antiviral agent for treatment of viral diseases in aquaculture.

Assessment and correction of local antimicrobial protection factors in women with chronic recurrent vulvovaginal candidiasis and type 2 diabetes mellitus

Aim. Evaluation and correction of local antimicrobial protection factors in women with type 2 diabetes mellitus (DM 2) and chronic recurrent vulvovaginal candidiasis (rVVC). Materials and methods. The study involved 100 women undergoing outpatient follow-up at the age of 40.9±5.8 years, with a body mass index of 29.8±3.5. The period was 2022–2023. The examination included anamnesis, collection of anthropometric data, calculation of body weight, microscopic examination of smears from the cervical canal and vagina, Gram-stained before treatment, in the 1st and 3rd months after its completion, identification pathogenic microorganisms using PCR, colposcopy, ultrasound of the pelvic organs. Indicators of glycosylated hemoglobin in the subjects at the control points of the study: at 1 month the average values were 5.9±2.9%, at 3 months – 5.9±3.0%, the average value was 5.9±3.1%. Candida species were identified by the bacteriological method using Sabouraud dextrose agar (growth of colonies of fungi of the genus Candida in an amount of more than 103 CFU/ml). Immunological methods for studying antimicrobial protection factors included studying the number of neutrophil granulocytes (NG) on the surface of the mucous membranes of the vulva and vagina, their phagocytic and NBT-reducing activity in a latex test. Randomization of patients into groups: group 1 included 50 (50%) women with DM 2 who, as part of complex therapy for rVVC received the drug Genferon® 1 intravaginal suppository of 500 thousand units 2 times a day for 10 days and fluconazole 150 mg orally three times with an interval of 72 hours at the first stage of treatment; group 2 included 50 (50%) women who received therapy with fluconazole 150 mg three times with an interval of 72 hours. The maintenance anti-relapse course therapy in both groups included the use of fluconazole 150 mg once a week for 6 months. In group 1st anti-relapse therapy was supplemented by the administration of the drug Genferon® 1 intravaginal suppository of 500 thousand units at night 3 times a week for 3 months, after which the vaginal microbiota was corrected for 2 weeks using vaginal suppositories, containing Lactobacillus acidophilus in an amount of at least 108 CFU. In group 2nd, basic anti-relapse antimycotic therapy was not accompanied by the prescription of any immunomodulatory or probiotic drugs. Results. The etiological agents of rVVC in women of late reproductive age in 79% (n=79) и 28% (n=28), respectively, were C. albicans and C. glabrata, which during the period of acute of the disease were detected by culture in the vaginal discharge at more than 104 CFU/ml. Chronic recurrent course of vulvovaginal candidiasis in women with DM 2 was characterized by exacerbations 4 or more times a year, accompanied by the corresponding clinical picture: white or yellowish-white cheesy discharge from the genital tract, itching or burning in the anogenital area, discomfort in the external genital area, dyspareunia, dysuria, decreased phagocytic activity of NG in vaginal secretions by 25.8%, impairment of their spontaneous and latex-induced NBT-reducing activity by 35.2%, functional reserve by 23.0% relative to reference values. The use of the drug Genferon® as part of complex therapy for rVVC contributed to a decrease in the number of Candida spp. in 1st and 3d months of observation after completion of anti-relapse therapy, normalization of cellular factors of innate immunity of the mucous membranes, faster resolution of the clinical manifestations of an episode of the disease. The decrease in the number of relapses over a 12-month observation period compared to the control group was also facilitated by an increase in the protective properties of the vaginal mucosa due to the restoration of the vaginal microbiota with the help of lactobacilli acidophilus. Conclusion. The etiological agents of rVVC in women of late reproductive age are C. albicans and C. glabrata. Subcompensated DM 2 is a risk factor for rVVC, which requires additional monitoring of microbiological parameters of the vaginal microbiota. rVVC in women with DM 2 is associated with an increase in the number of NG in the vaginal secretion, a decrease in their phagocytic activity and functional reserve compared to the reference values. Combination therapy of rVVC with topical recombinant interferon α2b, benzocaine and taurine in the formulation of Genferon® (suppositories) is an effective method to improve the clinical and immunological efficacy of therapy.

On the Possibility of Using the Topical form of the Recombinant Interferon alpha-2b Drug in the Prevention of Acute Respiratory Viral Infections in Organized Groups

Relevance. Respiratory infections are an urgent problem for organized groups, including military ones, since the transmission of pathogens by airborne droplets and household contact routes, physical and psychological stress on adaptive mechanisms contribute to the development of the epidemic process. There is no doubt that vaccination makes a significant contribution to the prevention of respiratory infections, but the contingent remains vulnerable to other pathogens against which there are no vaccines. Therefore, the search for new methods of non-specific prevention is necessary in maintaining the health of persons permanently residing in collectives. Aim. Evaluation of the possibility of using the topical form of the recombinant interferon α-2b drug for the prevention of acute respiratory viral infections in organized groups. Materials and methods. The work was carried out according to the methodology of a multicenter (3 Centers) double-blind controlled trial involving 3,235 people aged 18 to 22 years, who were divided into three groups: 1 gy. - received a recombinant interferon α-2b (Grippferon) drug 3 drops in each nasal passage 2 times a day in for two weeks; 2 g. - saline solution intranasally according to the same scheme; 3 g. - the volunteers did not receive anything. The frequency of respiratory infections was studied. Results and discussion. Medical monitoring of the study participants, which was carried out for 2 months, showed that in the groups from the Center 1, the incidence of acute respiratory diseases for 2 months in the main group (gr. 1) was 2.3 times lower than in the control (gr. 2) and the comparison group (gr. 3). In the Center 2 The data corresponded to the dynamics of Center 1: the incidence values were 2.4-2.5 times lower in Group 1. In Center 3, the values were 2.0-2.1, respectively. The epidemic effect of intranasal administration of the topical form of the drug recombinant interferon α-2b is due to its effect on the factors of mucosal immunity, which contributes to non-specific protection and increased body resistance against respiratory infections. Conclusions. The presented advantages of the recombinant interferon α-2b drug make it possible to draw attention to the clinical feasibility of its use for preventive purposes in organized groups, including military ones, from the position of high epidemiological effectiveness, both pre-exposure and post-exposure prevention of acute respiratory viral infections.

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.

Resting and activated bovine neutrophils and eosinophils differ in their responses to adrenergic agonists

Polymorphonuclear cells (PMN) provide a rapid response to infection and tissue damage and stress can modify these critical innate immune defences. The study of adrenergic receptor (AR) expression and function in bovine PMNs is limited but both neutrophils and eosinophils express numerous AR genes but differ significantly in their expression of individual AR genes. A flow cytometric technique was developed to differentiate between bovine neutrophils and eosinophils so both neutrophil and eosinophil responses to adrenergic agonists could be analysed. Neutrophils and eosinophils displayed significantly different changes in CD11b, L-selectin, and CD44 expression when activated by bovine serum opsonized zymosan and recombinant bovine interferon gamma. The responses of activated and resting neutrophils and eosinophils were then compared following stimulation with endogenous adrenergic agonists, epinephrine (E) norepinephrine (NE), and synthetic agonists targeting α1-, α2-, or β-ARs. Both resting and activated neutrophils and eosinophils displayed differences in iROS, CD44, and L-selectin expression following stimulation with E and NE. Resting neutrophils displayed pro-inflammatory responses to both E and NE, while resting eosinophils displayed a pro-inflammatory response to only NE. No single synthetic adrenergic agonist fully recapitulated responses observed with either E or NE and responses to adrenergic agonists were dose-dependent. In conclusion, bovine eosinophils and neutrophils responded to multiple adrenergic agonists by altering expression of proteins involved in immune surveillance and pro-inflammatory responses. Significant differences in neutrophil and eosinophil responses to adrenergic agonists are consistent with their differences in AR gene expression. This highlights the importance of analysing separately these two PMN subpopulations when investigating the effects of either endogenous or synthetic AR agonists.

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.

Interferons and interferon-related pathways in heart disease.

Interferons (IFNs) and IFN-related pathways play key roles in the defence against microbial infection. However, these processes may also be activated during the pathogenesis of non-infectious diseases, where they may contribute to organ injury, or function in a compensatory manner. In this review, we explore the roles of IFNs and IFN-related pathways in heart disease. We consider the cardiac effects of type I IFNs and IFN-stimulated genes (ISGs); the emerging role of the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) pathway; the seemingly paradoxical effects of the type II IFN, IFN-γ; and the varied actions of the interferon regulatory factor (IRF) family of transcription factors. Recombinant IFNs and small molecule inhibitors of mediators of IFN receptor signaling are already employed in the clinic for the treatment of some autoimmune diseases, infections, and cancers. There has also been renewed interest in IFNs and IFN-related pathways because of their involvement in SARS-CoV-2 infection, and because of the relatively recent emergence of cGAS-STING as a pattern recognition receptor-activated pathway. Whether these advances will ultimately result in improvements in the care of those experiencing heart disease remains to be determined.

Pharmacodynamic of Recombinant Human Interferon Alpha-2b Nasal Drops and Effective Prophylaxis Against SARS-COV-2 Infection.

The recombinant human interferon alpha-2b (IFN-α2b) nasal drop formulation (Nasalferon) was studied as prophylaxis for SARS-CoV-2. Healthy volunteers between 19 and 80 years of age received 0.5 million international units of IFN in one drop (0.05 mL ) in each nostril, twice a day, for 10 consecutive days. The nondetection of SARS-CoV-2 by real-time polymerase chain reaction was the primary outcome variable. Several IFN-α biomarkers, including intranasal gene expression and innate immune effector activity, were increased in participants who received intranasal IFN-α2b. The study included 2,930 international travelers and 5,728 persons who were their close contacts. The subjects were treated with Nasalferon in January 2021, and 9,162 untreated travelers were included as controls. COVID-19 rate in treated subjects was significantly lower than in untreated subjects (0.05% vs. 4.84%). The proportion of travelers with COVID-19 decreased from 60.9% to 2.2% between December 2020 and February 2021. Furthermore, 1,719 tourism workers also received Nasalferon, and no cases of SARS-CoV-2 infection were detected, whereas 39 COVID-19 cases (10.6%) were reported in 367 untreated subjects. The main adverse events associated with the use of intranasal IFN-α2b were nasal congestion, headache, and rhinorrhea. Our prophylactic health interventions study demonstrates that the daily administration of Nasalferon for 10 days decreases the risk of developing COVID-19 in healthy volunteers. [Figure: see text].

Certification of a pharmacopoeial reference standard for potency testing of recombinant interferon α-2b

SCIENTIFIC RELEVANCE. Potency testing of recombinant interferons requires a reference standard. The availability of International Standards (ISs) that are commonly used to assess the quality of recombinant interferons is currently limited. Therefore, the quality of interferons should be assessed using pharmacopoeial reference standards (RSs) certified using ISs (if available).AIM. This study aimed to certify a pharmacopoeial RS for potency testing of recombinant interferon α-2b.MATERIALS AND METHODS. The potency determination involved comparing the inhibition of the virus-induced cytopathic effect observed in cell culture with the candidate RS and the WHO International Standard for Human rDNA-derived interferon α-2b. The study used A-549 and MDBK cell lines with encephalomyocarditis (EMC) and vesicular stomatitis (VSV) viruses. The authors followed the requirements of the State Pharmacopoeia of the Russian Federation (Cell-Culture Bioassays for Interferon Products, OFS.1.7.2.0002.15). The results were recorded using instrumental and visual methods. The calculations used mathematical statistics. To factor in the influence of intermediate precision, the authors applied Student’s t-test.RESULTS. The authors developed a certification programme and procedure and certified the pharmacopoeial RS for recombinant human interferon α-2b (rhIFN α-2b) to identify the corresponding medicinal products (by virus neutralisation) and test their potency (by antiviral activity in cell culture). Upon receipt, the candidate RS was verified for compliance with regulatory specifications. According to the test results, the potency of the candidate pharmacopoeial RS was 4.47×108 IU/mL, and the expanded uncertainty was 8.12×107 IU/mL, with a coverage factor of k=2 and a confidence level of 95%. The pharmacopoeial RS for rhIFN α-2b was considered to have the same shelf-life period as the corresponding medicinal products if stored at a temperature not higher than –40 ºC. When thawed, the pharmacopoeial RS for rhIFN α-2b can be stored at a temperature of 2–8 ºC for up to 1 month.CONCLUSIONS. Upon certification for potency testing, the pharmacopoeial RS for rhIFN α-2b has been included in the Register of Reference Standards of the Russian Pharmacopoeia as Recombinant Human Interferon α-2b (Potency) FSO.3.2.00455. The introduction of this pharmacopoeial RS will help to conduct the identification and potency testing of all Russian interferon α-2b products at an appropriate scientific and methodological level. As a result, Russian pharmaceutical manufacturers and quality control agencies will no longer depend on imported ISs.

Intralesional interferon alpha-2b as a novel treatment for periocular squamous cell carcinoma in horses.

To determine the safety and efficacy of perilesional human recombinant interferon alpha-2b (IFNα2b) for treatment of periocular squamous cell carcinoma (PSCC) in horses. Eleven horses (12 eyes) with PSCC were enrolled in this prospective clinical study with owner consent. Systemically healthy horses were included in the study following confirmation of PSCC via biopsy. Every two weeks for a maximum of six treatments, horses were sedated and perilesional injection of IFNα2b (10 million IU) was performed. Tumors were measured prior to each injection and at one, three, and 12 months after treatment completion. A greater than 50% reduction in tumor size was considered positive response to treatment (i.e., partial or complete response). Development of anti-IFNα2b antibodies was assessed using serum samples obtained after treatment initiation and compared with treatment responses. Antibody concentrations were analyzed using a mixed model. Statistical significance was considered p < 0.05. Each horse received four to six perilesional injections of IFNα2b. Five of 12 eyes (4/11 horses) responded to treatment. Two of five eyes showed complete resolution of gross PSCC. No systemic adverse effects were seen. Local swelling occurred during treatment protocol in 6/11 horses but resolved without intervention. All horses developed serum anti-IFNα2b antibodies. There was no evidence of statistical difference in antibody concentration between responders and non-responders. Perilesional administration of IFNα2b was found to be well-tolerated in horses with PSCC, and induced tumor regression in 42% of treated eyes. Treatment failure appears unrelated to the development of IFNα2b antibodies.