Recombinant Interferon Research Articles

IMMUNE STATUS OF HYPOTROPHIC CALVES WHEN USING DRUGS BASED ON RECOMBINANT INTERFERONS

The article presents the results of studying the effect of drugs based on recombinant interferons on the body of hypotrophic calves in terms of changes in their immune status. Forty (40) newborn calves were selected for the experi-ment. The animals of the experimental group I (n=10) were administered the complex drug interamin at a dose of 1 ml/10 kg of animal weight, the animals of group II (n=10) were administered the drug Biferon-B according to a similar scheme. Group III (n=10) consisted of hypotrophic calves without using drugs. Group IV was represented by normotrophic calves. It was found that the use of the studied drugs had a positive effect on the completion rate of the adaptive processes of the formation of cellular and humoral factors of nonspecific defense of the body of hypotrophic calves.

LEVEL OF EXPRESSION OF THE INTERLEUKIN-6 GENE IN PIGS AGAINST THE BACKGROUND OF ETIOTROPIC THERAPY OF GASTROINTESTINAL PATHOLOGY IN COMBINATION WITH SPECIES-SPECIFIC RECOMBINANT CYTOKINES AND WHEN VACCINATED AGAINST PCV-2

The study presents the data on the level of gene expression of the interleukin-6 (IL-6g) gene when administering the drugs Proautovak and Feronamine, based on recombinant interferons, to pigs with diarrheal syndrome against the background of etiotropic therapy; and to swine when vaccinated against porcine circovirus with the drug Circoflex. The analysis of the data obtained showed a decrease in the expression of the IL-6 gene when using the drugs Proautovak and Feronamine against the background of treatment with Quinocol. When vaccinating, the use of the drug Proautovak led to a decrease in the expression of the IL-6 gene, while the drug Feronamine had no effect.

Влияние рекомбинантного интерферона α-2b на цитокинпродуцирующую активность клеток иммунной системы у детей с COVID-19

The clinical efficacy of recombinant IFN-α2b in therapy for COVID-19 in children is the basis for studying the parameters of the cytokine production activity of immune system cells and will allow to optimize antiviral therapy regimens. Objective. To study the effect of recombinant IFN-α2b on serum IFN-α and IFN-γ concentrations and their synthesis by immune system cells in children with COVID-19. Materials and methods. Peripheral blood samples from 100 patients aged 1 to 17 years (1–7 years – 50 people, 8–17 years – 50 people) with a moderate course of COVID-19 were examined. Patients in the study group received recombinant IFN-α2b as Вклюpart of complex therapy. Patients in the comparison group received antiviral therapy with Arbidol®. Multiplex analysis was used to determine serum IFN-α and IFN-γ concentrations and the level of their synthesis by immune system cells. The enzyme-linked immunosorbent assay (ELISA) was used to determine serum concentrations of antibodies to IFN-α. Results. Combination therapy with recombinant IFN-α2b in children of both age groups led to an increase in serum IFN-α concentrations compared to baseline values prior to treatment, in children in the control group and patients with COVID-19 after treatment with Arbidol. There were no significant changes in serum IFN-α and IFN-γ concentrations and their synthesis in intact and PHA-stimulated cells in children of both age groups during treatment with Arbidol. Serum concentrations of antibodies to IFN-α during treatment with recombinant IFN-α2b did not depend on the age of children and remained within the reference range. Conclusion. A significant increase in serum IFN-α concentrations and restoration of its synthesis level induced by PHA to reference values indicate that the use of recombinant interferon medications with their antiviral and immunomodulatory effects should become an integral part of COVID-19 therapy. Key words: children, COVID-19, recombinant interferon IFN-α2b, serum IFN-α concentrations, IFN-α synthesis levels

Comparative Study of Commercial Media to Improve GMP Manufacturing of Recombinant Human Interferon β- 1a by CHO Cells in Perfusion Bioreactor

Comparative Study of Commercial Media to Improve GMP Manufacturing of Recombinant Human Interferon β- 1a by CHO Cells in Perfusion Bioreactor

Next-generation version of recombinant human interferon-beta: A molecular dynamic simulation study

The ability of recombinant human interferon beta-1b (IFNβ-1b) to elicit the production of anti-IFNβ-1b antibodies becomes a significant issue for interferon therapy in patients. Mitigating the immunogenicity issues requires understanding the stimulating factors in observed immune responses. Among multiple factors, epitopes might be the main reason for the observed immunogenicity of this drug. In this study, we investigate the ability of epitopes in IFNβ-1b to bind to the MHC class II receptors. We applied point mutations to reduce the binding affinity of the existing epitopes of IFNβ-1b to MHC class II molecules. Several bioinformatics methods were applied to compare the binding affinity of both native and mutated epitopes to the MHC class II receptors. We identified three residues, Tyrosine 3, Tyrosine 132, and Glycine 162, that if altered to Leucine, Leucine, and Glutamate, respectively, leads to a decline in the binding affinity of IFNβ-1 b's epitope to the MHC class II receptors.This suggests that we can decline the binding affinity of therapeutic IFNβ-1b in silico by introducing those point mutations.

IMPACT OF INTERFERON-CONTAINING DRUGS ON PRO- AND ANTIOXIDANT STATUS IN NEWBORN PIGLETS

The article presents the results of studies on the impact of biferon-S, containing recombinant porcine interferons alpha and gamma, and prostimul, which contains a recombinant type I cytokine and vitamins A, E and C, on the pro- and antioxidant status of piglets under conditions of an industrial pig breeding complex. It has been found that the use of drugs for the newborn piglets helps to reduce the lipid peroxidation and endogenous intoxication, manifested by a decrease in the number of malondialdehyde, peptides of medium molecular weight and indicators of endogenous intoxication. This also contributes to a decrease in the number of stable nitric oxide metabolites involved into the oxidative stress reactions and an increase in the activity of antioxidant enzymes. A more essential positive effect on the LPO-AOD system in piglets has been exerted by prostimul, which, in addition to the type I cytokine, contains vitamins A, E and C. They increase the antioxidant status of the body, enhancing the antioxidant activity of the blood plasma, protecting the cells of the immune system from oxygen-dependent types of apoptosis, they potentiate the efficacy of interferons as well.

The use of recombinant interferon alpha-2b with antioxidants in the treatment of relapses of herpes simplex

The use of recombinant interferon alpha-2b with antioxidants in the treatment of relapses of herpes simplex

Оценка эффективности методов профилактики COVID-19 у беременных

Objective. To assess the effectiveness of different preventive measures for novel coronavirus infection in pregnant women. Patients and Methods. This study included 125 pregnant women hospitalized with moderate to severe laboratory-confirmed SARS-CoV-2 infection between September and November 2021 (the fourth pandemic wave), and 175 pregnant women who were not infected with COVID-19 during the same period. All women in these two groups were comparable for gestational age (II–III trimesters, 24–39 weeks), age (20–40 years), social status, parity, body mass index, and had no known COVID-19 risk factors. Results. Our findings revealed that vaccination 3-5 months before pregnancy (OR = 4.12; 95% CI 1.28–13.27; χ2 = 0.022), inconsistent use and/or non-timely replacement of face masks (OR = 5.71; 95% CI 2.83–11.51) were associated with the increased risk of COVID-19 in the second and third trimesters of gestation. It was showed that systematic (once in the morning at 24–48-hour intervals) intranasal administration of recombinant interferon alpha-2b (IFN-α; Grippferon) as compared with a single application after exposure to COVID-19 reduced the disease incidence rate and there was no evident risk of illness (OR = 0.08; 95% CI 0.05–0.14; 19.2% vs 74,3%, p < 0.001). This can be explained by the fact that women were mostly infected in unpredictable conditions (e.g., 29.2% of pregnant women were infected from family members, 23.9% had unknown source of exposure). The use of umifenovir, not currently authorised for the medication-assisted prevention of COVID-19 in pregnant women, and rectal administration of IFN-α suppositories did not reduce the disease incidence rate. Rectal use of IFN-α suppositories by pregnant women off-label increased the incidence (32.0 vs 15.4%, p = 0.001) and risk of developing novel coronavirus infection (OR = 2.58; 95% CI 1.48–4.50). Conclusion. There is a need to improve awareness among pregnant women about the mandatory and timely vaccination against COVID-19 during pregnancy and the importance of strict adherence to wearing face masks. Increased efforts should be made to monitor and inform pregnant women about the use of only authorised medication-assisted preventive measures of SARS-CoV-2 infection, such as intranasal administration of recombinant IFN α-2b (Grippferon). During the epidemic rise in COVID-19 cases, the systematic intranasal administration of recombinant interferon-based medication Grippferon (once in the morning at 24–48-hour intervals) is recommended for pregnant women. Key words: pregnancy, COVID-19, incidence rate, vaccination, recombinant interferon alpha-2b, Grippferon, prevention

USE OF PREPARATION OF RECOMBINANT INTERFERONS FOR CORRECTION OF OVULATORY FUNCTION IN OVARIES OF DAIRY COWS

The article presents the material on studying the efficacy of the preparation of recombinant interferons containing granulocyte-macrophage colony-stimulating factor (GM-CSF) for the prevention of the post-estral uterine bleeding in dairy cows, accompanied by a retardation in ovulation of the dominant follicle. It has been found that a single administration of the drug containing GM-CSF in its composition to cows at a dose of 5.0 ml during insemination, contributes to the ovulatory functional recovery in 86.7% of animals, and reduces the period from calving to fertilization by 27.7 – 37.8 days. The introduction of the recombinant interferon preparation to cows provides activation of the cellular link in the immune defense, normalization of the blood hormonal-cytokine profile, recovery of the ovulatory function in the genital gonads. This is manifested by an increase in the serum lysozyme activity – by 29.5%, leukocyte phagocytic activity – by 12.4%, phagocytic index – by 38.8%, phagocytic number – by 58.8%, progesterone – by 59.9%, estradiol – by 60.0%, testosterone – by 59.3%, DHEA-S – by 56.8%, IL-4 – by 42.5%, IL-10 – by 53.4%, with a decrease in the content of IL-2 – by 40.4% and IL-1β – by 37.2%.

Эффекты влияния рекомбинантного интерферона α-2b на фенотип субпопуляций нейтрофильных гранулоцитов пациентов с COVID-19

Investigation of molecular mechanisms associated with interferon (IFN) production and receptor function of neutrophil granulocytes (NGs) in COVID-19 is highly relevant because it can be promising in the search for new therapeutic strategies targeting NGs and their reactivity to restore and strengthen the innate immune response against SARS-CoV-2. Objective. To assess the effects of recombinant IFN-α2b on the phenotype of CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119–, and CD16+IFNα/βR1+CD119+ NGs from peripheral blood of patients with COVID-19 in an in vitro experiment. Patients and methods. We analyzed blood samples from 31 patients with a mean age of 61 years (range: 57;71 years) with moderate COVID-19. We assessed the number of CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119–, and CD16+IFNα/βR1+CD119+ NGs, receptor density (FC 500, ‘Beckman Coulter,’ USA), phagocytic activity of NGs before and after incubation with recombinant IFN-α2b. We also measured serum levels of several cytokines, including IFNα, IFNγ, IL-6, IL-8 (ELISA, ‘Vektor-Best’ LLC). The control group comprised 22 adult healthy individuals with a mean age of 58 years (range: 57; 70 years). Results. Patients with moderate COVID-19 demonstrated low serum levels of IFNα and IFNγ along with elevated levels of IL-6 and IL-8. We observed transformation of 3 phenotypes among NG subpopulations: CD16+IFNα/βR1–CD119+, CD16+IFNα/βR1+CD119-, and CD16+IFNα/βR1+CD119+. We observed positive remodulating effects of recombinant IFN-α2b on the number and phenotype of NG subpopulations and their phagocytic activity in our in vitro experiment. Conclusion. Recombinant IFN-α2b demonstrated positive effects in in vitro experiments; therefore, it can be considered in the future as a potential therapeutic tool for moderate COVID-19. Restoration of type I IFN might be an effective treatment option for COVID-19, because it can promote faster virus elimination, restore normal functioning of the IFN system, and have positive regulatory effects on the phenotype of NG subpopulations. Key words: neutrophil granulocytes, subpopulations, COVID-19, phenotype, recombinant interferon α2b

Preparation and Evaluation of rhINF-α-2b Sodium Hyaluronate Cross-Linked Porous Microspheres: Characterization, Sustained-Release Properties, and Antitumor Activity.

Recombinant human interferon α-2b (rhINF-α-2b), like most proteins, has several shortcomings such as relatively short half-life, low therapeutic index, high circulating drug fluctuations, and rapid degradation which could hinder its effective delivery. Novel electrostatic spray and ion exchange drug-loading techniques were combined to formulate rhINF-α-2b sodium hyaluronate cross-linked porous sustained-release microspheres (rhINF-α-2b-SHCPM), a protein delivery system. The different properties of rhINF-α-2b-SHCPM including the physicochemical nature, in vitro release behavior, and antitumor activity were evaluated. The loading rate (10.31 ± 0.94%) and encapsulation efficiency (89.09 ± 2.37%) of rhINF-α-2b-SHCPM produced acceptable values. The in vitro cumulative release rate of rhINF-α-2b-SHCPM within 24 h was also 86.26 ± 2.11% with a much better sustained release effect. Thus, the half-life (10.763 h) and retention time (14.067 h) of rhIFNα-2b-SHCPM were significantly prolonged with enhanced bioavailability (43,198.387 ng/L*h) and decreased peak concentration (15,266.4 ngL-1) compared with the free rhIFNα-2b protein (0.912 h, 0.952 h, 34,749.048 ng/L*h, and 48,870.2 ngL-1, respectively). The in vitro anti-proliferative activity and in vivo tumor inhibitory rate of rhIFNα-2b-SHCPM also increased by 90 and 55.86%, respectively, compared with the free rhIFNα-2b solution. The findings significantly supported a well-developed protein delivery system with improved sustained release, acceptable bioavailability, and increased antitumor activities. Graphical Abstract.

Functions of Traditional Chinese Medicine Combined with Recombinant Human Interferon α2b in Cervical Intraepithelial Neoplasias Patients.

Cervical cancer is a common malignant neoplasm in women, and its incidence is increasing year by year. This study explored the effects of traditional Chinese medicine combined with recombinant human interferon α2b in cervical cancer patients. 178 cervical intraepithelial neoplasias (CIN) combined with high-risk HPV-positive patients from June 2017 to August 2020 were divided into the study group (n = 89 cases) and the control group (n = 89 cases) by the random number table method. Patients in the control group were treated with recombinant human interferon α2b, and the study group was treated with traditional Chinese medicine (TCM) on the basis of the control group. After treatment, the recurrence rate in the study group was significantly decreased while the human papillomavirus (HPV) negative conversion rate was significantly increased. 3 months after treatment, the TCM symptom scores in the study group were lower than in the control group. Moreover, serum levels of inflammatory factors decreased in both groups, and the decrease was more significant in the study group. After treatment, the ultrasound parameters were significantly decreased in the study group than in the control group. In conclusion, traditional Chinese medicine combined with recombinant human interferon α2b in cervical cancer patients could effectively improve the negative conversion rate of HPV infection, the level of inflammatory factors, reduce the degree of cervical erosion, and enhance the immunity of patients with high safety and significantly improve the quality of life.

Delayed induction of type I and III interferons mediates nasal epithelial cell permissiveness to SARS-CoV-2

The nasal epithelium is a plausible entry point for SARS-CoV-2, a site of pathogenesis and transmission, and may initiate the host response to SARS-CoV-2. Antiviral interferon (IFN) responses are critical to outcome of SARS-CoV-2. Yet little is known about the interaction between SARS-CoV-2 and innate immunity in this tissue. Here we apply single-cell RNA sequencing and proteomics to a primary cell model of human nasal epithelium differentiated at air-liquid interface. SARS-CoV-2 demonstrates widespread tropism for nasal epithelial cell types. The host response is dominated by type I and III IFNs and interferon-stimulated gene products. This response is notably delayed in onset relative to viral gene expression and compared to other respiratory viruses. Nevertheless, once established, the paracrine IFN response begins to impact on SARS-CoV-2 replication. When provided prior to infection, recombinant IFNβ or IFNλ1 induces an efficient antiviral state that potently restricts SARS-CoV-2 viral replication, preserving epithelial barrier integrity. These data imply that the IFN-I/III response to SARS-CoV-2 initiates in the nasal airway and suggest nasal delivery of recombinant IFNs to be a potential chemoprophylactic strategy.

Development of highly stable and de-immunized versions of recombinant alpha interferon: Promising candidates for the treatment of chronic and emerging viral diseases

Human interferon alpha (hIFN-α) administration constitutes the current FDA approved therapy for chronic Hepatitis B and C virus infections. Additionally, hIFN-α treatment efficacy was recently demonstrated in patients with COVID-19. Thus, hIFN-α constitutes a therapeutic alternative for those countries where vaccination is inaccessible and for people who did not respond effectively to vaccination. However, hIFN-α2b exhibits a short plasma half-life resulting in the occurrence of severe side effects. To optimize the cytokine's pharmacokinetic profile, we developed a hyperglycosylated IFN, referred to as GMOP-IFN.Given the significant number of reports showing neutralizing antibodies (NAb) formation after hIFN-α administration, here we applied the DeFT (De-immunization of Functional Therapeutics) approach to develop functional, de-immunized versions of GMOP-IFN. Two GMOP-IFN variants exhibited significantly reduced ex vivo immunogenicity and null antiproliferative activity, while preserving antiviral function. The results obtained in this work indicate that the new de-immunized GMOP-IFN variants constitute promising candidates for antiviral therapy.

Effect of immune drugs to treat acute viral nasopharyngitis

The task in treating acute nasopharyngitis (ANP) deals with reducing the disease symptoms and the risk of complications. The lack of reliable antiviral drugs makes it important to search for appropriate medicines among other pharmacotherapeutic groups.The study involves a comparative analysis of the efficiency and estimates potential: the recombinant interferon α2b and the compound containing fungal β-D-glucans used in treat ANPThe studies involved patients with ANP from 18 to 55 years old. As many as 152 people were examined including the following: 38 were practically healthy people (group 1); and 114 patients wuth ANP: 38 people (group 2) was subject to a standard therapy (vasoconstrictor nasal drops, nasal cavity irrigation using 0.1% Miramistine solution, gargling using the Furacilin solution); forty people (group 3) were administered application of intranasal interferon α2b of 105IU, it was delivered with a spray into each nasal passage twice a day; 36 people (group 4) were administered an immunotropic drug containing β-D-glucans orally twice a day. The duration of drug administration lasted 7 days. Polymerase chain reaction (PCR) was used to identify the ANP etiological factor. Concentrations of cytokines IL-1β, IL-1ra were estimated using enzyme immunoassay (ELISA) technique. Clinical efficiency was assessed through score approach. The following symptoms were taken into account: general malaise, sore throat, character of nasal discharge, and the difficulty of nasal breathing. The results of the study were analyzed using parametric and nonparametric statistical methods. In 60.0% the nasal secretions of patients revealed RV. The distribution of cytokine concentrations in nasal secretions in group 1 indicated that the concentration of IL-1β was in the range of 20.0-25.0 pg/ml, and the concentration of IL-1ra was about 1250.0-2500.0 pg/ml. Developing ANP stimulated an increase in IL-1β concentration up to 30.0-70.0 pg/ml in nasal secretions of patients without affecting IL-1ra concentrations. On day 7 of treatment, the cytokine concentrations among the patients treated using the immunotropic drugs were the same as in the group of healthy individuals. There were no significant changes in cytokine production on day 7 in the group of patients undergoing the standard treatment. Application of proposed immunobiological medicines to ANP does not result in overproduction of proinflammatory cytokine IL-1β in nasal secretion. This confirms that these drugs are promising in the treating strategy including reduction of the risk of developing complications.

Evaluation of the Anti-Viral Activity of Human Recombinant Interferon Lambda-1 against SARS-CoV-2.

The antiviral activity of recombinant human IFN-lambda type 1 (IFNλ-1) against culture strain of SARS-CoV-2 virus was determined by infecting a highly sensitive VeroE6 coronavirus cell culture after preincubation test (the cell monolayer was incubated with 4-fold dilutions of IFNλ-1 in a concentration range of 0.16-42,500 ng/ml in a culture medium for 12 h at 37°C) and without preincubation (simultaneous addition of different concentrations of IFNλ-1 and SARS-CoV-2 infection in a dose of 102 TCID50). The created recombinant human IFNλ-1 demonstrated obvious antiviral activity against SARS-CoV-2 virus in vitro. In the tests with and without preincubation, IFNλ-1 exhibited significant activity, although somewhat lower in variant with simultaneous addition of IFNλ-1 and virus to the cell culture. It should be noted that the antiviral effect of IFNλ-1 was observed in a wide range of concentrations.

Investigation of the antiviral activity of the recombinant human interferon lambda 1 in human conjunctiva cell culture

The aim of the study was to determine the efficacy of recombinant human interferon lambda 1 (IFN-λ1) against human adenovirus serotype 5 in a culture of human conjunctival cells Chang conjunctiva clone 1-5c-4. Material and methods. The study design consisted of three experimental schemes, reflecting a prophylactic and two options for a therapeutic and prophylactic treatment regimen (with the constant presence of the virus in the culture medium and with its removal after adsorption). The antiviral activity of IFN-λ1 was determined by the number of viable cells after exposure to the virus (MTT test). Results and discussion. It has been established that IFN-λ1 has antiviral activity against human adenovirus in vitro under a prophylactic and therapeutic-prophylactic scheme of administration at an infection dose of 1 and 10 TCID50 (50% tissue culture infectious dose), but not at an infection dose of 100 TCID50. The antiviral effect of the use of IFN-λ1 in a therapeutic and prophylactic regimen at an infection dose of 1 TCID50 was comparable to that of IFN-α. At the same time, both interferons did not have a toxic effect on the cell culture even at a concentration of 84 and 58 μg/ml, respectively. The antiviral activity and the absence of cytotoxic action provide the basis for further study of the possibility of development of based on IFN-λ1 drug for eye conjunctiva viral diseases treatment.

The Effects of Chinese Herbal Decoction Combined with Recombinant Human Interferon α2b on MRI Imaging, Tumor Markers, and Immune Function in Patients with Renal Cell Carcinoma.

Background To explore the clinical efficacy of traditional Chinese medicine decoction combined with recombinant human interferon α2b in the treatment of renal cell carcinoma (RCC). Methods 68 RCC patients were divided into the control group and treatment group (n = 34). The control group was treated with recombinant human interferon α2b, and the treatment group was treated with traditional Chinese medicine decoction on the basis of the control group. The clinical efficacy and life quality were observed. At the same time, the changes of immune function before and after treatment were compared. Results After one course of treatment, the effective rate and clinical benefit rate of the treatment group were higher than those of the control group. The Karnofsky score in the treatment group was better than that in the control group. In improving the immune function, the treatment group was better than the control group in increasing CD3+ and CD4+ and reducing CD8+. Conclusion Traditional Chinese medicine decoction combined with recombinant human interferon α2b has a good effect on the treatment of RCC. It can not only improve the common clinical symptoms of patients but also improve the quality of life and cellular immune function of patients.

Validation of a test procedure for determination of specific activity of human recombinant interferon beta (Infibeta®) using various cell/virus combinations

Interferon beta preparations have demonstrated efficacy in the treatment of multiple sclerosis. One of the most important quality attributes that support efficacy and safety of interferon beta preparations is specific antiviral activity. Interferon beta activity is determined by the biological test method which can lead to an erroneous final result due to the system uncertainties that contribute to the overall uncertainty. Accurate assessment of specific activity plays an important role in adequate determination of the product’s therapeutic dose, therefore the test method standardisation and validation are of particular relevance. The aim of the study was to validate a test procedure for assessing specific activity of human recombinant interferon beta preparations, using various cell/virus combinations, as illustrated by the example of Infibeta®. Materials and methods: WISH, Vero, A-549, and MDBK cells in combination with mouse encephalomyocarditis virus were used in the study. The testing was performed using the biological test method based on the interferon ability to suppress virus-induced cytopathic effects in cell cultures. The results were processed using methods of mathematical statistics and the GraphPadPrism, Statistical 2/0 software package. Results: the paper compares the results of specific activity determination of Infibeta® (interferon beta-1b) using various cell/virus combinations. The dose-response curves were used to compare the test results. It was demonstrated that all the tested cell lines could be used in the biological test procedure for determination of interferon beta specific activity. However, the best results were obtained with A-549, WISH cells in combination with mouse encephalomyocarditis virus. The following validation characteristics were determined in the cell/virus systems: specificity, linearity, precision, and robustness. Conclusions: the study validated the test procedure that enables measurement of interferon beta specific activity in the range of 4.8–11.2 IU/mL at a satisfactory accuracy level, which guarantees reliable test results. The study demonstrated robustness, intermediate precision, and reproducibility of the test procedure.

Investigation of Cytotoxic Effects of Recombinant Human Interferon Lambda-1 and Its Pegylated Form on Human Conjunctival Epithelial Cells

Currently, there are no efficacious, all-purpose antiviral medicines for the treatment of ocular surface infections caused by viruses. At the same time, type III interferons demonstrate high potency for histological barriers, such as the conjunctiva. Modification of protein molecules in native products can significantly improve their pharmacodynamic properties. Thus, it seems reasonable to develop antiviral medicines based on interferon lambda (IFN-λ1) and its pegylated form (PEG IFN-λ1).The aim of the studywas to evaluate thein vitrocytotoxic effect of recombinant human IFN-λ1 and its pegylated form onChang conjunctiva clone 1-5c-4human conjunctival cells.Materials and methods:PEG IFN-λ1 was obtained by the electron beam immobilisation method. A normal human conjunctival cell lineChang conjunctiva clone 1-5c-4was used for cell cultivation. The MTT test was used to assess the cytotoxic effect. Cell proliferative activity was studied by measuring microelectrode impedance. Ultrastructural changes were assessed by electron microscopy. Statistical processing was performed using the Statistica 10.0 software package.Results:IFN-λ1 (37 μg/mL) and PEG IFN-λ1 (42 μg/mL) had no significant cytotoxic effect on the human conjunctiva cell culture and the cell proliferative activity. The analysis of ultrastructural changes demonstrated that IFN-λ1 activated metabolic processes in the cells, and PEG IFN-λ1 promoted differentiation and keratinisation of epithelial cells and led to modification of the cell membrane. A ten-fold increase in IFN-λ1 and PEG IFN-λ1 concentration (to 370 μg/mL and 420 μg/mL, respectively) reduced the cell viability by 15–20% as compared to the intact control.Conclusions:the study results demonstrated that IFN-λ1 and PEG IFN-λ1 could be used as active pharmaceutical ingredients in the development of medicines for the treatment of conjunctival viral infections.