[Development and preservation of specific T-cell immunity after COVID-19 or vaccination against this infection].

Aim evaluation of specific T-cell immunity against SARS-CoV-2 in primary and secondary response to virus antigens by screening method. MATERIALS AND METHODS: Patients were tested 11.5 months after COVID-19 and 610 months before and after vaccination. Healthy volunteers were screened before, 26 times during the vaccination course, and 68 months after revaccination with the Sputnik V vaccine. IgG and IgM antibodies to SARS-CoV-2 were detected by ELISA using commercially available kits (Vector-Best, Russia). Antigenic (AG) activation of T cells in the fraction of bloods mononuclear cells was assessed by IFN- production after AG stimulation in the wells of plates from ELISA kits intended for detection of antibodies against SARS-CoV-2. Data were processed by MS Excel and Statistica 10.0 software. RESULTS: AG-specific T cells were detected in 88.5% of vaccinated healthy volunteers, half of whom were found to have T cells appearing earlier than antibodies to AG. After 6-8 months, the level of AG activation decreases. Following the revaccination, the level of AG activation of memory T cells in vitro increases within six months in 76.9100.0% of vaccinated subjects. On the contrary, after COVID-19, 86.7% of individuals had in their blood the AG-specific T cells with high activity at the time of vaccination. The activity of T cells recognizing the RBD domain of the SARS-CoV-2 S protein and the proportion of individuals who had these cells in their blood increased after the vaccination of reconvalescents. CONCLUSION: T-cell immunity against SARS-CoV-2 antigens has been shown to persist for 6 months after illness. In vaccinated individuals without history of COVID-19, such duration of the preservation of AG-specific T cells in blood was only achieved after the revaccination.

[Assesment of specific T-cell immunity to SARS-CoV-2 virus antigens in COVID-19 reconvalescents].

INTRODUCTION: The development of the COVID-19 pandemic has stimulated the scientific research aimed at studying of the mechanisms of formation the immunity against SARS-CoV-2. Currently, there is a need to develop a domestic simple and cost-effective specific method suitable for monitoring of T-cell response against SARS-CoV-2 in reconvalescents and vaccinated individuals. AIM: Development of a screening method for evaluation specific T-cell immunity against SARS-CoV-2. MATERIALS AND METHODS: Total 40 individuals who had mild to moderate COVID-19 and 20 healthy volunteers who did not have a history of this disease were examined. The presence and levels of IgG and IgM antibodies to SARS-CoV-2 were identified in participants sera by ELISA using the diagnostic kits from JSC Vector-Best (Novosibirsk, Russian Federation). Antigenic stimulation of mononuclear cells was carried out on commercial plates with adsorbed whole-virion inactivated SARS-CoV-2 antigen (State Research Center of Virology and Biotechnology VECTOR Novosibirsk, Russian Federation). The concentration of IFN- was measured in ELISA using the test systems from JSC Vector-Best (Novosibirsk, Russian Federation). The immunophenotyping of lymphocytes was performed on a flow cytometer Cytomics FC500 (Beckman Coulter, USA). Statistical data processing was carried out using the Microsoft Excel and STATISTICA 10 software package. RESULTS: Stimulation of mononuclear cells isolated from the peripheral blood with whole-virion inactivated SARS-CoV-2 antigen fixed at the bottom of the wells of a polystyrene plate showed a significantly higher median response in terms of IFN- production in 40 people who had history of COVID-19 compared to 20 healthy blood donors (172.1 [34.3575.1] pg/ml versus 15.4 [6.925.8] pg/ml, p 0.0001). There was no difference in median IFN- levels in supernatants collected from unstimulated mononuclear cells from COVID-19 reconvalescents and healthy donors (2.7 [0.411.4] pg/ml versus 0.8 [0.023.3] pg/ml, p 0.05). The overall sensitivity and specificity of this method were 73% (95% CI 5888%) and 100% (95% CI 100100%), respectively, at a cut-off of 50 pg/ml. CONCLUSION: The developed method for assessment of the cellular immune response to SARS-CoV-2 can be used as a screening method for monitoring the T-cell response in a population against a new coronavirus infection in recovered people.

Features of the Humoral Response to Infection, Vaccination, and Revaccination during COVID-19.

IgM and IgG antibodies to the SARS-CoV-2 virus are detected in subjects who have recovered from COVID-19; IgM antibodies persist in a 1/3 of infected subjects up to 12 months from the moment of the disease, while IgG antibodies are present in the vast majority of cases (97%; medium and high levels antibodies were registered in 85% of cases). By the 12th month, 40% of those who recovered still have a very high level of IgG antibodies to the S-protein (>500 BAU/ml). In the feces, urine, and blood serum of patients with long-term persistent IgM antibodies, no coronavirus antigens were detected. After vaccination with the Gam-COVID-Vac vaccine, IgG antibodies to the S-protein are detected in 100% of cases and remain at a high level for 4 months, by the 5-6th month, the level of antibodies decreases. During revaccination, the level of IgG antibodies to S-protein reaches high values earlier than during primary vaccination, and remains high for 4 months (observation period). The blood sera of recovered and vaccinated patients have a high virus-neutralizing activity (at least 1:80), while its level is somewhat higher in recovered patients.

Dynamics of Antibodies to Various Antigens of the SARS-CoV-2 Coronavirus in Patients with Confirmed COVID-19 Infection.

The presence of IgG and IgM antibodies in the venous blood of 76 patients with confirmed COVID-19 infection was determined by ELISA using Russian test systems. Different levels of IgM antibodies to N-protein and receptor binding domain of the Spike protein (RBD) were revealed. The dynamics of IgG antibodies to the whole virion antigen and recombinant antigens showed high values on weeks 4-5 of the disease. The level of IgG antibodies to Nprotein remained low throughout the observation period. The characteristic dynamics of IgG measured using test systems with sorbed whole virion or recombinant spike proteins reflects the duration of the disease.

Analysis of the Pharmacokinetics of Suppository Forms of Bacteriophages.

Pharmacokinetics of suppository forms of bacteriophages was studied on male Chinchilla rabbits. Suppositories with various composition of bacteriophages were administered once per rectum to rabbits, and the presence of phage particles was estimated in the blood, urine, and feces over 24 h. Pharmacokinetic study showed that the phages were detected in the blood, urine, and feces at various terms of the experiment irrespective of the size of viral particles, which confirmed the possibility of their systemic effects after rectal administration. Thus, the use of suppository form of bacteriophages can ensure the presence of phage particles even in infection foci that cannot directly contact with the preparation.

[COMPLEX IMMUNOGLOBULIN PREPARATION FOR THERAPY OF PERTUSSIS IN YOUNG CHILDREN].

AIM: Study the possibility of inclusion of complex immunoglobulin preparation (CIP) pos- sessing specific activity against pertussis exotoxins into complex therapy of pertussis infection in young children. MATERIALS AND METHODS: -2 groups of children with.pertussis younger than 3 years were examined. The main group (50 individuals) received CIPper os - 1 dose 1 - 2 times per day for 5 days, comparison group (34 children) received only basic therapy. Evaluation of clinical effectiveness of CIP was carried out, the content of anti-pertussis class G antibodies and total IgE in patients were studied. RESULTS: A good clinical effectiveness of the preparation was shown, as well as immune modulating activity against humoral immune response to pertussis infection. CONCLUSION: The detected positive effect of CIP on pertussis -course in children has indicated a principally novel use of this per oral drug form.

POLYMER TRACK MEMBRANES FOR ATMOSPHERIC PRESSURE FIELD EXTRACTION OF IONS FROM LIQUID SOLUTIONS.

An approach to the generation of gas phase ions by field extraction from liquid solutions has been investigated. The method uses a polymer membrane with nano-size channels as an interface between the liquid and the atmospheric pressure gas. Ions are produced by dissociation in the polar solvent and secondary ion-molecular reactions in the solution, which fills the channels of the membrane. Field extraction of the ions from the channels is stimulated by pulses of the electric discharge between the membrane and an adjacent electrode in the gas. The gas-phase ions are removed from the extraction zone by air flow and are detected by mass spectrometry. Possibilities of the membrane interface for generation of gas phase ions have been demonstrated from mass spectral investigation curried out for angiotensin II, gramicidin S and cytochrome C solutions. The current kinetics of the membrane ion source has been investigated to elucidate the mechanism of the ion extraction.

[Intranasal administration of immunoglobulins--perspectives for use in medical practice].

Effectiveness of topical use of immunoglobulines in respiratory infections along with preparations for parenteral use is discussed. Immunoglobuline preparations for intranasal use (drops, spays, aerosols) should take place among preparations intended for prevention and treatment of influenza, parainfluenza, respiratory syncitial virus infection and others. Potential to intranasal use of complex immunoglobulin preparation containing polymeric and monomeric antibodies of different isotypes is also discussed.

[Determination of protease activity in blood and in microorganisms].

For determination of protease activity it is possible to use immunoglobulins. Since proteolytic products apparently do not retain substrate antigenic determinants, it is possible to use ELISA methodsfor monitoring for enzymatic process. ELISA determination of functional activity of specific IgA1-protease has been applied not only for detection of this enzyme, but also for measurement of its inhibition constants. Fixed on a micropanel IgG may be used for evaluation of total proteolytic activity. Depending on pH values, it is possible to measure activity of neutral, alkaline and acid proteases. This approach has allowed to estimate total proteolytic activity of neutral proteases of serum. Measurement of a total level of serum pepsinogene activity can have diagnostic importance in gastroenterology, due to decisive contribution of pepsinogen I to the measured activity.

[Protein content in blood plasma and its changes in patients of the neurological critical care unit]. / Soderzhanie belkov v plazme krovi i ego dinamika u bol'nykh neiroreanimatsionnogo otdeleniia.

Twelve blood plasma proteins were studied in 120 patients with isolated traumatic and nontraumatic cerebral hemorrhage. At all stages of the study a marked imbalance of blood proteins occurred which was independent of the etiological factor. Patients with craniocerebral trauma exhibited the phenomenon of the atypical response manifesting with low concentration of acute-phase proteins. The levels of the complement component C5 and transferrin in patients with cerebral vascular aneurysm rupture are shown diagnostically significant while follow-up of blood transtiretin can show the trend in the metabolic processes in patients with cerebral hemorrhage in critical condition.

[Evaluation on monkeys of reactogenicity and effectiveness of the complex immunoglobulin preparation formulation]. / Otsenka na obez'ianakh reaktogennocti i éffektivnosti lekarstvennykh form kompleksnogo immunoglobulinovogo preparata.

Experiments on 75 monkeys, 20--healthy and 75--with diarrhea, showed that ready medicinal forms of the complex immunoglobulin preparation in tablets, capsules or in a dried form (vials) were safe, well tolerated, clinically and bacteriologically effective. When used simultaneously with antiparasitic treatment, this preparation, irrespective of the kind of its ready medicinal form, showed high curative effect (80-90%). The diarrhea ceasing was accompanied by the gradual normalization of intestinal microbiocenosis (an increased concentrations of Escherichia coli and Bifidobacterium, a decreased amount of opportunistic bacteria). The complex method of the treatment of diarrhea in monkeys (antibiotics, trichopol, phage and probiotics) made it possible to achieve curative effect only in 60% of cases, which required the additional course of treatment, namely the use of rehydration therapy.

alpha1-acid glycoprotein possesses in vitro pro- and antiinflammatory activities.

We studied the effects of alpha1-acid glycoprotein on tumor necrosis factor-alpha (TNF-alpha) and interleukin-10 (IL-10) production and lymphocyte response to phytohemagglutinin in cultured peripheral blood mononuclear leukocytes from 6 healthy donors. We observed 2 opposite responses to alpha1-acid glycoprotein: first, stimulation of TNF-alpha and IL-10 production and inhibition of lymphocyte proliferation, and second, suppression of cytokine production and stimulation of lymphocyte proliferation. In cell cultures isolated from 4 of 6 donors, the TNF-alpha/IL-10 ratio remained unchanged after addition of native alpha1-acid glycoprotein, but some fractions isolated by chromatography on concanavalin A-Sepharose changed this parameter. These changes were most pronounced after treatment with fraction C enriched with molecules with incomplete (biantennary) carbohydrate chains. The mechanisms of alpha1-acid glycoprotein-induced effects on peripheral blood mononuclear leukocytes are discussed.

[Early evaluation of blood loss by the profile of blood proteins in patients with injuries]. / Ranniaia otsenka ob"ema krovopoteri po profiliu belkov plazmy krovi u bol'nykh s travmoi.

The concentrations of positive (C-reactive protein, orosomucoid, haptoglobin, ceruloplasmin) and negative (transtiretine, alpha 2-macroglobulin, transferrin) markers of acute phase response were measured in 55 victims with injuries to hollow organs on day 1 of hospitalization. By visual intraoperative evaluation of blood loss, the patients were divided in 2 groups: with blood loss under 1 liter (n = 32, group 1) and more than 1 liter (n = 23, group 2). Acute phase response was more manifest in group 1. Blood loss affected the levels of orosomucoid and haptoglobin, which is explained by functional characteristics of these blood proteins. Orosomucoid and haptoglobin tests were highly sensitive, specific, and accurate in evaluation of blood loss in patients with hollow organ wounds (87.5, 90, 89% and 80, 80, 80%, respectively). The content of these proteins equal to no more than 1 g/liter during the first day after the injury indicates blood loss higher than 1 liter with high probability.

The solvent shells of cluster ions produced by direct electric field extraction from glycerol/water mixtures.

Electric field extraction of gaseous negative ions directly from water/glycerol solutions by use of a track membrane technique was investigated. The distributions of numbers of solvent molecules in the extracted cluster ions for different compounds were obtained. It is shown that the extraction mechanism is a direct field-stimulated evaporation of cluster ions from liquid, with a subsequent loss of several solvent molecules in the vacuum. For relatively simple ions a good correspondence of results was obtained with a continuous medium model. It was found that the number of solvent molecules in a cluster shell, for more complicated ions such as amino acids, is significantly greater than that for halide ions or ions of simple organic acids. An increase in the number of solvent molecules in the case of amino acid negative ions is rationalized in terms of the existence of several charged groups, each of which gives an additional contribution to the cluster shell.

[Electrolaser therapy on the Iarilo device in patients with chronic chlamydial prostatitis]. / Elektrolazernaia terapiia na apparate "Iarilo" u bol'nykh khronicheskim khlamidiinym prostatitom.

Multiple-modality treatment (antibiotics, immunocorrectors, vascular medication, physiotherapy) for chronic chlamydial prostatitis (CCP) was conducted in 141 patients. They were assigned to two groups which differed only by physical methods applied. Group 1 patients (n = 84) were exposed to electrolaser radiation (Yarilo unit), group 2 patients (n = 57) were treated by conventional physiotherapeutic methods. The advantages were reported in the group treated simultaneously with He-Ne laser introduced into the urethra, infrared laser, electrostimulation (Yarilo unit). Perineal pain and dysuria ceased in 58.9 and 9.8%, 62.1 and 13.2% of patients from group 1 and 2, respectively. Erection recovered in 90% of group 1 patients. In group 2 this percentage was 50%.

[The production of immunochemical reagents for the determination of C-reactive protein by latex agglutination]. / Poluchenie immunokhimicheskikh reagentov dlia opredeleniia C-reaktivnogo belka metodom lateksnoi aggliutinatsii.

A new simple and convenient method for the detection of C-reactive protein (CRP) in patients' blood sera is based on latex agglutination making use of new compounds obtained by conjugation of IgG fraction containing goat antibodies to CRP with various latex particles. The sensitivity of the methods is 3 mg/ml, this permitting its wide clinical application.

[Structure of carbohydrate chains of molecular forms of alpha1-acidic glycoprotein from ascitic fluid from patients with stomach cancer]. / Struktura uglevodnykh tsepei molekuliarnykh form alpha1-kislogo glikoproteina iz astsitnoi zhidkosti bol'nykh rakom zheludka.

The immunochemically pure alpha 1-acid glycoprotein (aAGP) from ascitic fluid of patients with stomach cancer was separated by chromatography on Con A-Sepharose into Con A-nonbound (aAGP-1, 43, 5 kDa, 70%) and Con A-bound (aAGP-2, 41.5 kDa, 24% and aAGP-3, 40.0 kDa, 5%) forms, differing in the monosaccharide composition. Comparative study of structures of their N-carbohydrate chains with the aid of the HPLC of fluorescence-labelled oligosaccharides showed that the molecular forms differ by the ratio of the di-, tri-, and tetraantennary carbohydrate N-chains of a complex type. The molecular forms of aAGP differ from nAGP by amounts of Le(x)-fragments and agalacto-oligosaccharides.

[Molecular forms of alpha-1-acid glycoprotein in human blood serum. Differences in levels of di-, tri-, and tetra-antennary N-linked carbohydrate chains]. / Molekuliarnye formy alpha-1-kislogo glikoproteina syvorotki krovi cheloveka. Razlichiia v soderzhanii di-, tri-, i tetraantennykh N-uglevodnykh tsepei.

alpha 1-Acid glycoprotein isolated from healthy individuals blood was separated on Con A-Sepharose into three fractions: non-bound (AGP-1, 84%, 43.5 kDa), Con A-bound (AGP-2, 14%, 41.3 kDa), and Con A-tightly bound (AGP-3, 2%, 39.6 kDa). Amino acid compositions of these fractions were similar but carbohydrate ones differed. HPLC analysis of 7-amino-4-methylcoumarin derivatives of the oligosaccharides in combination with their sequential exoglycosidase digestion showed that AGP-1, AGP-2, and AGP-3 have the same set of oligosaccharides and differ only by their proposition. A minor quantity of agalacto-oligosaccharides (with a terminal GlcNAc residue) was identified.